15 September 2022, Volume 48 Issue 17
    

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  • LI Yanjie, GAO Jing, JIANG Yanjun, ZHOU Liya
    Food and Fermentation Industries. 2022, 48(17): 1-7. https://doi.org/10.13995/j.cnki.11-1802/ts.030343
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    Selective monooxygenation of inert hydrocarbon bonds is a kind of chemical reaction with exploratory significance in organic synthesis. At present, chemical method and enzymatic method are mainly used for catalysis. Chemical catalysis method often has harsh reaction conditions, while enzymatic catalysis method is relatively mild and has high regional and enantioselectivity. Unspecific peroxidase (UPO, EC 1.11.2.1) from Agrocybe aegerita can carry out complex monooxygenation reaction with H2O2 as co substrate. However, unspecific peroxidase (AaeUPO) as a heme-dependent enzyme, is easy to inactivate under high concentration of H2O2. In situ H2O2 production catalyzed by enzyme can provide lower concentration H2O2 for AaeUPO continuously and stably, and the process is mild and efficient. It is one of the effective ways to avoid adding high concentration H2O2. Therefore, taking AaeUPO and glucose oxidase (GOx) as the research objects, a cross-linked enzyme aggregate (combi-CLEAs) was prepared, and a method for enzyme catalyzed in-situ production of H2O2 and chiral alcohols was established. In this cascade system, GOx oxidizes glucose while reducing O2 to H2O2, and then AaeUPO catalyzes hydroxylation reaction with H2O2 as co substrate. In order to obtain combi-CLEAs with good performance, the effect of preparation conditions on the recovery of combi-CLEAs enzyme activity was investigated. The results showed that the optimum preparation conditions of combi-CLEAs were 6:1 (U:U) of GOx:AaeUPO , the volume ratio of precipitant isopropanol to crude enzyme solution was 2:1, the volume fraction of crosslinker glucan aldehyde (200 kDa) was 15%, and the mass concentration of BSA was 40 μg/mL and the crosslinking time was 3 h. At this time, the recovery of enzyme activity of combi-CLEAs was 87.43% and the specific enzyme activity was 63.12 U/mg. When it is used to catalyze the hydroxylation of ethylbenzene, under the optimal catalytic conditions of pH 7.0 and 30 ℃, continue to optimize the concentration of glucose to make GOx use glucose to stably and continuously generate H2O2 required for AaeUPO. When the concentration of glucose is 6 mmol/L, the conversion of ethylbenzene can reach 99.9%, and the ee value of (R)-phenylethanol is greater than 99.9%. In the reusability experiment of ethylbenzene catalyzed by combi-CLEAs, it can still maintain 67.20% ethylbenzene conversion after 8 cycles. The hydroxylation of different substrates was catalyzed by combi-CLEAs. The results show that the catalytic performance of combi-CLEAs is better than AaeUPO-CLEAs with H2O2, and has a good application prospect.
  • LIU Song, HAN Xuyue
    Food and Fermentation Industries. 2022, 48(17): 8-15. https://doi.org/10.13995/j.cnki.11-1802/ts.030130
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    Laccase is a multi-copper oxidase, which has important applications in food, textile, environmental remediation, and papermaking. To improve the fermentation level of laccase, Bacillus subtilis laccase (CotA) was expressed in Escherichia coli BL21 (DE3), and the effects of ribosome binding site, induction expression condition, and co-expression of lysis protein on its expression efficiency were investigated. The results showed that the recombinant strain with the optimal ribosome binding site (CotA-RBS5) achieved 1 236 U/L of intracellular CotA, 1.75 times higher than that of before optimization. Based on the orthogonal experimental, the optimal condition of induction expression was as follows: the final concentration of 2 mmol/L Cu2+ and 0.1 mmol/L IPTG were added when the biomass OD600 reached 0.8; then, induction condition was performed at 30℃. Under the optimal condition, the intracellular enzyme activity of the recombinant strain increased to 7 926 U/L, 5.40 times higher than that of before optimization; meanwhile, the extracellular enzyme activity reached 4 984 U/L. Co-expression of lysis protein E promoted cell lysis. With the expression of cleavage protein E was induced at 3.5 of OD600, the extracellular CotA activity reached 10 283 U/L as it fermented to 30 h. The results will provide important basic data for the industrial production of laccase.
  • CONG Huihui, ZHANG Yanan, NIU Dandan, ZHAO Jihua, WANG Zhengxiang
    Food and Fermentation Industries. 2022, 48(17): 16-20. https://doi.org/10.13995/j.cnki.11-1802/ts.032442
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    Bacillus naganoesis pullulanase (PulA) is a starch debranching enzyme that has achieved large-scale production in China.However, its enzymatic properties (especially saccharification pH) are inconsistency with that of glucoamylase in starch saccharification, which effects on the conversion rate of starch to glucose.The characteristics of an acidic mutant, LoPulA, from PulA, and its application value in the production of glucose from starch were studied.The results showed that the maximal activity of LoPulA was at pH 4.5 and 60 ℃ and kept 96.40% of the maximal activity at pH 4.3, which was greatly consistent with the optimal saccharification pH with glucoamylase.In starch saccharification experiments, the DX value of the sugar solution could reach 96.66% with the synergistic hydrolysis of LoPulA and glucoamylase, which was 1.40% higher than that of PulA (wild type) and glucoamylase, thereby indicating LoPulA of great application potential in high glucose syrup preparation from starch.
  • LI Min, ZHU Li, ZHAN Xiaobei
    Food and Fermentation Industries. 2022, 48(17): 21-28. https://doi.org/10.13995/j.cnki.11-1802/ts.030855
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    Agrobacterium sp. WS-8E3T produced low-molecular-weight β-1,3-glucan was obtained by adaptive domestication of Agrobacterium sp. ATCC 31749. Before optimization, the yield of low-molecular-weight β-1,3-glucan by the bacterium used glycerol as a carbon source was 1.648 g/L. To obtain high-yield oligosaccharides with lower cost, the fermentation conditions were optimized. The optimal initial glycerol concentration was 20 g/L, after replenished 20 g/L glycerol when the residual glycerol content was about 10 g/L. The optimal initial ratio of compound nitrogen source was 4 g/L of yeast extract and 3 g/L of ammonium sulfate, with a liquid volume of 75 mL/500 mL and pH of 6.3. In 7 L fermenter, during the growth period of the bacteria, the optimal pH was 7.0, and the glycerol was 20 g/L. During the oligosaccharide-producing period, the pH was 6.3, and the glycerol was 30 g/L. The final yield of oligosaccharides reached 8.03 g/L, which was 386.7% higher than that of before optimization. The results of FT-IR analysis and monosaccharide composition was consistent with curdlan, and the degree of polymerization was 18-22. The single-strain fermentation strategy was used to significantly increase the yield of low molecular weight β-1,3-glucan for the first time, which provide a theoretical basis for further low-cost and large-scale fermentation production of low molecular weight β-1,3-glucan.
  • DAI Yue, WU Dan, ZHENG Pu, CHEN Pengcheng
    Food and Fermentation Industries. 2022, 48(17): 29-34. https://doi.org/10.13995/j.cnki.11-1802/ts.030589
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    Alternan is a novel functional oligosaccharides which has good prebiotic properties. The content of alternate sugars is less in nature and mainly prepared by microbial fermentation and enzyme conversion. Currently, low activity of alternansucrase prevents its application in largescale. To enhance the enzyme expression, the alternansucrase gene was synthesized and heterologously expressed in Pichia pastoris GS115 for the first time. After 72 h high-density fermentation in 3 L fermenter, the enzyme activities in supernatant reached 34.5 U/ml, which was about 12 times than that of shake flask fermentation, and reached the highest level of recombinant alternansucrase at present. The optimum pH and temperature of recombinant ASR were 6.0 and 55℃, Km and kcat were 31.97 mmol/L and 0.36 s-1, respectively. In addition, alternan was prepared by using sucrose or sucrose and other receptors as substrates at 40°C and pH 5.5. It was identified by TLC that both oligoalternan and polyalternan were produced in the catalytic products. The alteransucrase gene was successfully expressed in P. pastoris for the first time, which will provide a basis for the heterologous expression of alteransucrase and the large-scale preparation of alternan.
  • ZHANG Wen, WANG Fangting, PANG Jingcheng, SHI Xiangzhu, CHEN Bingdian, NI Li
    Food and Fermentation Industries. 2022, 48(17): 35-41. https://doi.org/10.13995/j.cnki.11-1802/ts.029713
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    The purpose of this study is to produce the probiotics of Bacillus subtilis BS08 with high viable count and high stability.Spray drying technology was used to select better wall materials from protein, carbohydrate and hydrocolloid protectant by single factor experiment.And, the number of viable bacteria and stability of bacterial powder was used as indicators to study the effect of the embedding order of anti-thermal protectant on the protective effect of BS08. Finally, the response surface methodology was applied to optimize the formulation of compound anti-thermal protectant.The results showed that the anti-thermal protectant composed of gelatin, arabic gum and trehalose from the inside to the outside could effectively protect BS08 in microcapsules.Through stability evaluation, it is estimated that the viable count of the product were >1014 CFU/g if storage for 1 year at 4 ℃, with the same order of magnitude.After response surface optimization, when the mass fraction of gelatin, Arabic gum and trehalose is 26%, 24%, and 24%, the maximum viable count of BS08 is 5.98×1014 CFU/g.
  • LAI Wenmei, TAN Shuyu, SHI Feng
    Food and Fermentation Industries. 2022, 48(17): 42-48. https://doi.org/10.13995/j.cnki.11-1802/ts.032011
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    4-Hydroxyisoleucine (4-HIL) has great potential in the treatment of type II diabetes. Isoleucine dioxygenase (IDO) can convert L-isoleucine (Ile) into 4-HIL. To improve the product of 4-HIL and reduce the by-product L-lysine (Lys) synthesis in recombinant Corynebacterium glutamicum, the Lys-OFF riboswitch was integrated into the upstream of dapA gene, the key gene of Lys synthesis pathway, and the D-RS strain was obtained. The strain could dynamically minimize the synthesis to Lys by the concentration of intracellular and extracellular Lys, and decreased the content of Lys by 46.7%. Secondly, the Ile-activated biosensor Lrp-PbrnFEN was used to control the expression of codon-optimized ido in D-RS strain, and the titer of 4-HIL was increased to 116.3 mmol/L. In order to further improve the titer of 4-HIL, the strong promoter PbrnFE7 was used to dynamically control the expression of odhI and vgb to enhance the supply of α-ketoglutarate and oxygen. Finally, a recombinant strain D-RS-0I7O7V with the 4-HIL titer up to 166.0 mmol/L and Lys content down to 6.5 mmol/L after fermentation in shake flasks was obtained. The dynamic regulation strategy provides a new idea for the efficient biosynthesis of 4-HIL.
  • CHENG Kun, LIU Rui, SUN Sijia, ZHAI Lei, ZHENG Jia, YAO Su
    Food and Fermentation Industries. 2022, 48(17): 49-56. https://doi.org/10.13995/j.cnki.11-1802/ts.029691
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    Bacillus aquiflavi 3H-10 was isolated from yellow water of strong-flavor Chinese Baijiu in Yibin, Sichuan province, China, and it had significant ability in producing protease and various flavor substances. High-throughput genome sequencing of strain 3H-10 was carried out and antimicrobial resistance genes, virulence factors genes, pathogenicity, antimicrobial susceptibility testing and virulence test in mice were analyzed to comprehensively evaluate the safety of the strain. The result showed that the genome of strain 3H-10 was a circular DNA with a size of 3 806 094 bp and the G+C content was 35.66 mol%. A total of 4 031 genes were predicted along with 81 tRNA, 27 rRNA and 16 sRNA. No potential antimicrobial resistance genes were detected in the genome of 3H-10 and the strain was sensitive to 10 common antibiotics affiliated with nine classes, indicating no risks of horizontally transferring antimicrobial resistance. No potential virulence genes were detected in the genome of strain 3H-10, and the virulence test demonstrated no toxic reaction or death in mice, implying that the strain 3H-10 was non-virulent or non-human pathogenic. It was concluded that strain B. aquiflavi 3H-10 possesses no safety risks in the aspect of antimicrobial resistance, virulence or pathogenicity.
  • LI Ruiying, ZHANG Yilin, YOU Chunping
    Food and Fermentation Industries. 2022, 48(17): 57-63. https://doi.org/10.13995/j.cnki.11-1802/ts.029676
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    The antioxidant capacity of several Lactobacillus strains was compared with Lactobacillus rhamnosus GG (LGG) strain in vitro. Lactobacillus rhamnosus B6 was selected with a more comprehensive antioxidant capacity, and the antioxidant capacity of its metabolites cultured in different media was compared. Then, the oxidative damage model of PC12 cells was established to determine the content of malondialdehyde and ROS after the treatment of probiotic metabolites, and to explore the effect of metabolic products of B6 on cellular oxidative stress. The results showed that the free radical scavenging rate of DPPH, ·OH and ·O2- of L. rhamnosus B6 were 63.23%, 99.38% and 26.35%, respectively, significantly higher than that of LGG. Compared with fermentation in MRS, DPPH free radical and ·OH free radical scavenging rates of metabolites obtained by B6 fermentation with skim milk significantly increased, and ·O2- free radical scavenging rates were similar between each other. The oxidative damage of PC12 cells reduced and the antioxidant capacity improved under oxidative stress with B6 metabolites fermented with skim milk.
  • HU Mengying, XU Haoyuan, HAN Jinpeng, ZHANG Lianzhong, NIU Ce, WANG Zhibin
    Food and Fermentation Industries. 2022, 48(17): 64-70. https://doi.org/10.13995/j.cnki.11-1802/ts.030332
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    To test the effects of Lactobacillus plantarum P-8 on the intestinal flora, antioxidant and anti-inflammatory capacity of mice for providing a basis for the development and utilization of a probiotic preparation of L. plantarum P-8, Fifty-two BALB/c male mice were randomly divided into blank control, low, medium and high-dose groups and administered with normal saline and different doses of probiotic preparation of L. plantarum P-8 for 30 d. The results showed that, compared with the control, there was no significant difference on the food intake, body weight and organ coefficient in each dose group(P>0.05), the levels of GSH, T-SOD and GSH-Px in the small intestine and liver tissues significantly increased (P<0.05) and the levels of MDA significantly decreased in the high-dose group (P<0.05) and the levels of GSH-Px significantly increased in the medium-dose group (P<0.05). The levels of IL-1β and IL-6 significantly decreased in the high-dose group (P<0.05). After administration, there was no significant difference in the number of microorganisms in blank control, low and medium-dose groups (P>0.05), the number of Lactobacillus and Bifidobacterium significantly increased in the high-dose group (P>0.05). 16S rDNA high-throughput sequencing results showed that, compared with the control, the abundance of Lactobacillus significantly increased in the low-dose group (P<0.05) and extremely increased in the medium and high-dose group (P<0.05), the abundance of Odoribacter significantly increased (P<0.05) and that of Muribaculum and Bacillus decreased in the high-dose group significantly (P<0.05). In summary, L. plantarum P-8 can improve the antioxidant capacity, reduce serum inflammation level, regulate the composition of intestinal flora and has a great application potential in the development of probiotic preparation.
  • ZHANG Jing, WU Ying, GU Shaobin, MA Jinliang, TIAN Pingping, ZHAO Lina, LI Xin, ZHANG Jie
    Food and Fermentation Industries. 2022, 48(17): 71-78. https://doi.org/10.13995/j.cnki.11-1802/ts.030314
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    Antimicrobial peptides are natural biological preservatives that inhibit the growth of food-borne pathogens. In order to obtain antimicrobial peptide from Bacillus coagulans CGMCC 9951, whole genome sequencing and bioinformatics analysis were used to mine the antimicrobial peptide synthesis gene cluster, and their expression and activity were verified. In order to study the antibacterial activity of the peptide, the fusion protein containing glutathione S-transferase (GST) tag was successfully constructed in Escherichia coli and verified by mass spectrometry. The results showed that two antimicrobial peptides were excavated by antiSMASH software. After comparison with National Center for Biotechnology Information (NCBI) database, Region 3 was found to be greatly similar with Amylocyclicin, and was selected for expression. It was found that mature Region 3 was composed of 64 amino acid residues with a molecular weight of 6 371.57 Da, which was positively hydrophobic protein. Homology of Region 3 showed cyclic antimicrobial peptide, and the tertiary structure was mainly α helix structure. Using Listeria monocytogenes as an indicator bacterium, the inhibition zone diameter was (15.2±1.4) mm, which confirmed that the expressed fusion protein had antibacterial activity. The successful expression of the fusion protein lays a foundation for the further study of B. coagulans CGMCC 9951 antimicrobial peptide.
  • WEI Bo, WANG Jie, CHEN Yixin, WANG Chunguang, CHEN Zhuojun, ZHANG Bolin, ZHU Baoqing
    Food and Fermentation Industries. 2022, 48(17): 79-86. https://doi.org/10.13995/j.cnki.11-1802/ts.030337
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    Real-time monitoring of microbial dynamics during fermentation is essential for wine quality control. A method could distinguish between dead and live microbes for Saccharomyces cerevisiae was developed, which combines the fluorescent dye propidium monoazide (PMA) with CELL-qPCR. It also could detect the quantity of microbes efficiently and rapidly without DNA extraction during wine fermentation. The method was optimized for cell wall breakage, PMA treatment concentration, PMA detection sensitivity and multiple conditions of sample pretreatment in wine environment. The results showed that the optimum method can accurately quantify 103-107 CFU/mL of the target strain in multiple matrices; when the concentration of dead microorganisms in the system was 104 times higher than that of live microorganisms, there was an error of 0.5 lg CFU/mL; the optimized PMA-CELL-qPCR method was used to monitor the dynamic changes of S. cerevisiae during the fermentation of Cabernet Sauvignon wine were consistent with the plate count method. In conclusion, the live bacteria quantification method developed for PMA-CELL-qPCR in S. cerevisiae wines is accurate in quantification and simple in operation, and can be used as a mean to accurately monitor microbial dynamics in wine and other fruit wines.
  • GUO She, XU Ming, ZHANG Can, ZHU Hanfang, CHEN Xia
    Food and Fermentation Industries. 2022, 48(17): 87-92. https://doi.org/10.13995/j.cnki.11-1802/ts.032080
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    Lysogenic phages of Lactobacillus spp. are widely distributed in dairy fermentation setting. Their special infection characteristics are somewhat hidden and arduous to detect. However, under certain conditions, these phages could cause lysis of fermenting lactic acid bacteria strains, and eventually lead to fermentation failure. Therefore, lysogenic phages have become one of the major safety hazards in dairy fermentation. The Lactobacillus fermentum IMAU32579 was induced and the inactivation effects of heat treatment and different chemical biocides were evaluated. The results showed that a bacteriophage named as LFP-VB1was successful induced from L. fermentum IMAU32579 after treatment with 1.9 μg/mL mitomycin C. Morphology analysis revealed that this bacteriophage strain belonged to the family of Siphoviridae. It was observed that bacteriophage LFP-VB1 was sensitive to heat treatment (completely inactivated 63℃ for 4 min). The inactivation effects of biocides were insignificant, 6.29 lg decrease was observed after treatment with 800 mg/L NaClO for 60 min. This study can provide some theoretical basis for the constructing of phage control measures.
  • LI Haoying, XU Yan, WANG Dong
    Food and Fermentation Industries. 2022, 48(17): 93-101. https://doi.org/10.13995/j.cnki.11-1802/ts.029964
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    Chinese traditional Jiuyao (the fermentation starter of Chinese traditional rice wine, Mijiu) has a wide range of sources and different fermentation characteristics. Exploring the key factors influencing the fermentation characteristics of different Jiuyao is helpful to control the production and improve the quality of Mijiu. Two different types of Jiuyao by natural culture, saccharification-type Jiuyao (SJ) and saccharification and fermentation-type Jiuyao (SFJ), were used for the fermentation of Mijiu. The key factors that may cause differences in the fermentation characteristics of different Jiuyao were explored. Based on correlation analysis and experimental verification of Mijiu fermentation, the results showed that key factors influencing the fermentation characteristics of Jiuyao was the acidic protease activity of Jiuyao, though the content and fermentation characteristics of yeast were also very important. The increase of acidic protease activity of Jiuyao could improve the fermentation performance by modifying the content of α-amino nitrogen required by yeast growth and thus affecting yeast proliferation at the initial stage of Mijiu fermentation. It could even convert a saccharification-type Jiuyao into a saccharification and fermentation-type one. Therefore, the activity of acidic protease in Jiuyao could be an important indicator for predicting the fermentation characteristics and the classification of Jiuyao. These results provide reference for the classification of Jiuyao as well as the control of Mijiu production.
  • SHI Getu, FU Jianhua, WU Yuan, CHEN Qiuyu, ZHENG Jiajing, LYU Jiaheng, LI Jiming, TANG Ke
    Food and Fermentation Industries. 2022, 48(17): 102-107. https://doi.org/10.13995/j.cnki.11-1802/ts.030789
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    Tannins, as important non-volatile matrix components of wine, can significantly affect the aroma perception of wine and then affect preferences of consumers. The model and mechanism of their influences on the orthonasal and retronasal perception of wine aroma are still unclear, which is a focus of wine flavor research. In this study, a three-alternative forced-choice (3-AFC) procedure, combined with the best estimate threshold values (BETs), was used to comprehensively evaluate the influence of tannins on the orthonasal and retronasal detection thresholds of six common and important aroma compounds in simulated wine. The results showed that tannins increased the orthonasal thresholds of 1-octen-3-one, β-damascenone, benzaldehyde and vanillin, among which the orthonasal thresholds of β-damascenone (from 7.2 μg/L to 25.4 μg/L) and vanillin (from 35.5 μg/L to 296.9 μg/L) had the most significant changes. The dominant factors of changes may be the hydrophobic mechanism of interaction between tannins and aroma compounds and the π-π stacking mechanism, and it is assumed that other physicochemical properties of the compounds such as polarity may also influence changes in the threshold. Besides, tannins increased retronasal thresholds of 1-octen-3-ol and vanillin and decreased retronasal thresholds of 1-octen-3-one, β-damascenone and benzaldehyde. Moreover, oral physiological factors such as saliva would complicate the study of the mechanism of tannins' influences on retronasal thresholds. It is worth noting that tannins had no significant influences on the orthonasal threshold of 1-octen-3-ol and both the orthonasal and retronasal thresholds of isoamyl acetate. Studies have shown that although tannins can reduce the release of most aroma during wine drinking, the effects on the release of some aromas were not significant and changes in aroma release do not necessarily cause sensory changes. Overall, tannins can alter the orthonasal and retronasal thresholds of aroma compounds. However, it is also worth noting that tannins had significantly different effects on the orthonasal and retronasal thresholds of the same compound, which was caused by the huge differences in the in vivo and in vitro environment, especially the influence of a large number of physiological factors involved in the in vivo environment, as well as the influence of perceptual interactions. The various molecular mechanisms involved in this still need to be further explored. This study will provide data and theoretical basis for the control of aroma perception in wine glasses and during wine drinking based on non-volatile matrix tannins.
  • YANG Yanli, YANG Yuhang, WANG Luyao, SUO Huayi, WANG Hongwei, ZHANG Yu
    Food and Fermentation Industries. 2022, 48(17): 108-115. https://doi.org/10.13995/j.cnki.11-1802/ts.029873
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    In order to explore the quality changes and clarify the best aging time during the fermentation and aging process of lemon wine, Eureka lemon was used as raw material to brew low-alcohol lemon wine. The volatile components, physicochemical characteristics, and the sensory qualities of lemon wine in different periods were determined and analyzed. The results showed that during fermentation and aging, the soluble solid content and pH decreased, while the titratable acid content increased, and the alcohol content increased at first and then decreased, reaching a peak of 4.27%vol at 28 d of aging. The color change of lemon wine in each period didn't show an obvious trend. A total of 109 volatile components were detected, including 15 esters, 25 alcohols, 36 terpenes, 5 acids, 14 aldehydes and ketones, and 14 other types. Besides, the sensory qualities of lemon wine in different stage were distinguishing. Based on the results, lemon wine aged for 28 days and 42 days perform better in terms of physicochemical characteristics and volatile components, but the wine aged for 28 days has better sensory quality.
  • WU Youna, WANG Yurong, YANG Yang, SHUANG Quan
    Food and Fermentation Industries. 2022, 48(17): 116-121. https://doi.org/10.13995/j.cnki.11-1802/ts.029627
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    To clarify the changes in microbial community structure during the fermentation of congee, high-throughput sequencing was used to analyze bacteria and fungi at different fermentation times of congee. The physicochemical characteristics and the correlation between microbial diversity and physicochemical indexes of congee were analyzed. The results showed that the dominant bacteria in congee during nine fermentation periods were mainly Lactobacillus, Kosakonia, Enterococcus, Ralstonia, Erwinia and Klebsiella. And the fungi were mainly Yarrowia, Dipodascus, Pichia. During the whole fermentation process, the relative content of dominant bacteria fluctuated obviously, and more bacteria participated in the fermentation than fungi, and Lactobacillus, Yarrowia and Dipodascus were always the dominant microorganism, and the acidity, soluble solids, protein and most amino acids of congee showed an upward trend. Correlation analysis showed that the Enterococcus and Pichia played an active role in the formation of most amino acids. The pH has an important influence on the fermentation of congee, therefore, controlling the acidity of fermentation plays a crucial role in the quality of fermented congee.
  • XI La, XIANG Fanshu, ZHANG Yan, ZHANG Haibo, GUO Zhuang
    Food and Fermentation Industries. 2022, 48(17): 122-128. https://doi.org/10.13995/j.cnki.11-1802/ts.028616
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    The bacterial community structure and function of Zha-chili from Tianmen region were analyzed by MiSeq high-throughput sequencing and PICRUSt software, respectively, and the lactic acid bacteria were isolated, identified and preserved by a pure culture method. Fifteen samples were divided into two clusters. The results showed that the bacteria in cluster I were mainly affiliated to Lactobacillus, with a relative abundance of 89.77%; the bacteria in cluster II were mainly affiliated to Klebsiella, Kosakonia, and Tetragenococcus, with relative abundances of 19.81%, 12.24%, and 10.89%, respectively. The two clusters were found to be highly significantly different (P<0.001) from one another by Mann Whitney test; meanwhile, a significant negative correlation observed between Lactobacillus and other bacterial genera such as Kosakonia and Pseudomonas (P<0.05). There were significant differences in 15 functions among the two clusters, but both of them were highly expressed in the transport and metabolism functions of amino acids and carbohydrates. The results of isolation and identification of lactic acid bacteria showed that L. plantarum, L. brevis, L. curvatus and L. sakei were contained in Zha-chili, and L. plantarum was the dominant Lactobacillus in Zha-chili from the Tianmen region.
  • LI Chunyan, XIONG Zhiqiang, WANG Guangqiang, SONG Xin, YANG Yijin, ZHANG Hui, AI Lianzhong, XIA Yongjun
    Food and Fermentation Industries. 2022, 48(17): 129-135. https://doi.org/10.13995/j.cnki.11-1802/ts.030574
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    Lactic acid bacteria (LAB) adjuncts have important impacts on the maturity and function of cheese. To improve the maturation characteristics of natural cheese, the fermentation characteristics of 24 LAB strains were evaluated, including acidification, autolysis, activity of aminopeptidase, proteolysis, lipolysis, salt tolerance, exopolysaccharide production activities and growth under low-temperature conditions. Then the cheese slurry rapid ripening model was applied to evaluate the influence of the selected strains on the cheese ripening. The results showed that the four strains (Lactobacillus fermentum AR497, Lactobacillus plantarum AR611, Lactobacillus salivarius AR612, and Lactobacillus salivarius AR809) had the potential to be adjunct cultures, with low acid production capacity, high autolysis and aminopeptidase activities, moderate protein hydrolysis and lipolysis activity, and the ability to adapt to high-salt and low-temperature environments during cheese ripening. The cheese ripening model results revealed that four adjunct cultures had no significant effect on the basic composition of the cheese slurry (pH, moisture, fat, and protein), and the four strains could proliferate well in cheese slurry without affecting the growth of starter cultures. Protein degradation analysis illustrated that four strains stimulated increased in the pH 4.6-soluble nitrogen and 12% trichloroacetic acid-soluble nitrogen contents in cheese slurry, among which AR809 had the most significant effect on promoting the breadth and depth of casein hydrolysis (P<0.05). Reversed phase-HPLC results showed that the types and content of hydrophilic peptides had an increasing trend as the ripening progressed. L. salivarius AR809 was able to increase the hydrolysis of casein to form peptides and had a higher number of peaks and peak areas compared with control. The above results indicate that AR809, as an adjunct culture, has good application potential in the ripening process of cheese.
  • CHEN Lele, SHI Haibo, SHEN Fangdi, WU Junkai
    Food and Fermentation Industries. 2022, 48(17): 136-142. https://doi.org/10.13995/j.cnki.11-1802/ts.032010
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    To analyze the changes of organic acids, free amino acids and volatile flavor compounds during the fermentation of coix seed inoculated with Lactobacillus pentosus AB-8, the molecular sensory science, artificial sensory evaluation, stability testing and multivariate statistical analysis were used. Results showed that after fermentation with L. pentosus AB-8 for 48 h, the contents of organic acids in coix seed increased from 380.98 mg/L to 5 440.37 mg/L. The contents of lactic acid changed significantly, reaching 3 412.67 mg/L, accounting for 62.73% of the total organic acids. The contents of total amino acids and essential amino acids of coix seed increased by 3.72 times and 5 times, respectively, compared with control. The kinds and contents of volatile flavor compounds in coix seed also increased after fermentation. The results of artificial sensory evaluation showed that the sour, sweet and umami were the main taste in coix seed fermentation with 48 h, while fruit and flower aromas were the most prominent aromas. In addition, the stability of coix seed increased, the absolute value of its zeta potential significantly reduced from 9.16 mV to 1.13 mV, and the particle size increased from 1 020 nm to 5 116 nm after fermentation. This research provides theoretical basis and data reference for the fermentation technology of coix seed.
  • YI Puhong, LI Qinping, ZHAO Caihong, DING Xiaoyan, WANG Ning, ZHU Tingheng, LI Ji
    Food and Fermentation Industries. 2022, 48(17): 143-149. https://doi.org/10.13995/j.cnki.11-1802/ts.030250
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    The high lipid content of kitchen waste has become one of the problems affecting its treatment and utilization. Through enrichment of samples from different environmental sources and screening on lipid degradation plates, a highly efficient lipid degradation bacterium strain JY34 was obtained and identified as Microbacterium esteraromaticum by 16S rDNA analysis. The lipid degradation rate of JY34 under different temperature, pH, inoculation amount and lipid content conditions were explored, and its lipase activity and lipid degradation rate in the wastewater of kitchen waste were evaluated. The results showed that the soybean oil degradation rate of JY34 was 98.78% at 30℃, pH 7.0, the inoculation amount of 2% and lipid content of 1% after fermentation for 3 d. The lipase activity in the crude enzyme solution of JY34 increased with temperature rising among in the range of 30-50℃, reaching 159.5 U/L at 50℃ after fermentation for 2 d. After JY34 strain was fermented in the kitchen waste lipid-containing wastewater for 3 d or 6 d, the lipid degradation rate reached 47.49% or 72.92%, respectively, which were 4.1 times and 1.63 times than that of control. In summary, strain JY34 has good adaptability to mesophilic condition and lipid degradation ability, so it will have a broad application prospect in the treatment of oily wastewater from kitchen waste.
  • ZENG Lin, WANG Zhaojun, HE Zhiyong, QIN Fang, ZENG Maomao, CHEN Jie
    Food and Fermentation Industries. 2022, 48(17): 150-158. https://doi.org/10.13995/j.cnki.11-1802/ts.031660
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    Heat-induced mixed gels were prepared with soybean protein isolate (SPI) and camellia seed protein (CPI) as raw materials. The rheological properties, texture, water holding capacity, and gel structure of the mixed gels were determined. The interaction between the two proteins and the mechanism that lead to the improvement of the mixture gel were explored based on the respective amino acid compositions of the two proteins, the solubility, electrophoresis, surface free sulfhydryl groups, disulfide bonds, surface hydrophobicity, solution particle size distribution, secondary structure, gel microstructure, etc. Results showed that when the compounding ratio of SPI and CPI was between 10:2 and 10:3, the gelling point temperature of the mixed protein decreased from 82.34 ℃ to 75.73 ℃, and the storage modulus value reached the maximum, which was 5 times that of pure SPI. Meanwhile, the gel strength reached the maximum, which was 2 times that of pure SPI. Moreover, the electron microscopy results showed that an appropriate proportion of CPI enabled the mixed protein to form a dense and uniform gel network structure. The results of electrophoresis, surface free sulfhydryl content, and disulfide bonds content showed that the two proteins interacted by disulfide bonds and was formed a complex between the subunits, which could reduce the surface hydrophobicity of the complex and effectively improve the solubility of SPI, it also could promote the gel network structure to be denser and more uniform. Furthermore, infrared spectroscopy results showed that the secondary structure of the mixed protein was changed from β-sheet to β-turn and random coil. Above results indicated that the improvement of the gel strength of soy protein by a certain proportion of camellia seed protein was mainly based on the interaction between the two proteins. A certain proportion of camellia seed protein had an enhancement effect on the gel strength of soybean protein, which provided a new thought and theoretical basis for the preparation of high gel plant-based mixed protein in the future.
  • LI Cili, GAO Yuan, ZENG Jianhua, SUN Bingyu, HUANG Yuyang, ZHU Xiuqing
    Food and Fermentation Industries. 2022, 48(17): 159-167. https://doi.org/10.13995/j.cnki.11-1802/ts.031387
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    In order to explore the effect of pH-shift induction on the dissociation-association behavior of soybean lipophilic protein(SLP)and the self-assembled nanoparticles, the SLP was treated by pH-shift inducted and the dissociation-association behavior of SLP and its self-assembled nanoparticles induced were characterized by thermal analysis and dynamic light scattering techniques. The results showed that the structure of SLP protein experienced unfolding, subunit dissociation and reaggregation during pH-shift induction. After about 10 h, more tyrosine and tryptophan were distributed on the surface of SLP with a stable and a loose three-dimensional conformation. The results of Fourier transform infrared spectroscopy and circular dichroism spectroscopy showed that pH-shift induction (10 h) changed the hydrophilic side chains on the protein surface and destroyed the hydrogen bonds and other forces acting on the secondary structure of the protein. Besides, it also caused the α-helix structure change to spin and increased the β-sheet content. The thermal analysis results showed that SLP had the maximum anti-degeneration ability at the isoelectric point. During the process of pH-shift induction, SLP nanoparticles self-assembled to form spherical nanoparticles with regular shape and uniform size, whose molecular morphology changed from disordered aggregation state to uniform particle size distribution. The SLP nanoparticles induced at pH 4 showed a stable single distribution system, and then exhibited good molecular flexibility and emulsifying properties.
  • KAN Jintao, GONG Shufang, WANG Yuanyuan, SONG Fei, SHEN Xiaojun
    Food and Fermentation Industries. 2022, 48(17): 168-174. https://doi.org/10.13995/j.cnki.11-1802/ts.028780
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    To meet the demand of the coconut processing industry for healthy, stable and fragrant food flavoring agent, gum Arabic and maltodextrin were used as wall materials in this experiment, and virgin coconut oil was used as core material to prepare coconut oil nano-microcapsules by complex condensation method. Fourier transform infrared spectroscopy, scanning electron microscopy, laser particle size analysis, thermogravimetric analysis, differential scanning calorimetry, oxidation stability analysis was used to characterize the physical and chemical properties of microcapsules, in vitro controlled release performance analysis and release mechanism exploration respectively. The results showed that the nano-microcapsules of coconut oil were successfully prepared by the complex condensation method. The nano-microcapsules had a spherical, irregular, multi-core and unfixed structure, and the average particle size was 378.07 nm. The nano-microcapsules had better thermal stability and oxidation stability. The in vitro sustained-release performance analysis showed that more than 65% of the core material was released in the simulated gastric fluid within 15 mins, which indicated that it could been shortly and rapidly released. The release mechanism of coconut oil nano-microcapsules in gastric juice was Fickian free diffusion mechanism. However, in intestinal fluid, it conformed to the Huguchi release model, which was the diffusion mechanism of the gel layer and skeleton dissolution release mechanism. The result provides a theoretical and technical basis for the application of coconut health and natural coconut flavor.
  • YAN Shanyao, CHI Hai, FENG Guangpeng, FANG Wenhong, ZHANG Shuping, HUANG Yanqing
    Food and Fermentation Industries. 2022, 48(17): 175-182. https://doi.org/10.13995/j.cnki.11-1802/ts.029526
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    Firstly, the enzymatic hydrolysis conditions were optimized by using single-factor (material-to-liquid ratio, amount of enzyme, temperature, time) and orthogonal test from defatted Antarctic krill powder in this work. Antarctic krill peptides with high-Fischer ratio (HF-AKP) were prepared by ultrafiltration and followed by activated carbon adsorption. Then the CCK-8 method was used to observe the effect of HF-AKP on the growth of human liver cancer cell line HepG2. Finally, the cell cycle distribution and apoptosis-related protein expression of HepG2 were detected by flow cytometry and ELISA methods, which made efforts to explore the effects of HF-AKP on HepG2 cells. The results revealed that enzymatic hydrolysis conditions of alkaline protease were as follows: the material-to-liquid ratio was 1:17.5 (g:mL) and the addition amount of enzyme was 5.0% for enzymatic hydrolysis at 55℃ for 6 h. The flavor protease optimal conditions were as follows: the addition amount of enzyme was 4.0% and enzymatic hydrolyzed for 3 h. After the second round hydrolysis, the degree of hydrolysis increased up to(17.4±0.2)%. Especially, after activated carbon adsorption, the Fischer ratio of defatted Antarctic krill peptides increased from (2.4±0.2) to (87.2±0.7) compared to AKP. HF-AKP had an inhibitory effect on the cell proliferation of the HepG2, which was positively dependent on dose- and time- manner. It also could block HepG2 cells in the G0/G1 phase. In addition, the mechanism of apoptosis may be related to the increase in Bax/Bcl-2 protein expression ratio and the decrease in NF-κB protein expression, so HF-AKP may have certain anti-tumor effects.
  • LI Conghu, CHEN Qiang, WU Shuangshuang, HUO Hongyue, CHENG Xu, WU Yan, DUAN Lian
    Food and Fermentation Industries. 2022, 48(17): 183-190. https://doi.org/10.13995/j.cnki.11-1802/ts.031888
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    The effects of concentration, temperature, and pH on the aggregation behavior of native and acylated collagen molecules were investigated by fluorescence spectrum technology. Results showed that the characteristic fluorescence peaks of native and acylated collagen molecules presented at 277 and 293 nm, which belonging to the typical peak of phenylalanine (Phe) and tyrosine (Tyr), respectively. Compared with native collagen, acylated collagen molecules peak had a wide range at 360-400 nm, belonging to the excited state of Tyr, indicating that the aggregation degree of acylated collagen molecules was larger than that of native one. With the increase of concentration, the aggregation degree of native and acylated collagen molecules increased which resulted in the gradual increase of fluorescence intensity. However, with the increase of temperature, the fluorescence quantum yield of Phe and Tyr decreased, and both native and acylated collagen molecules depolymerized until the random curl structure appeared. Therefore, the fluorescence intensity of native and acylated collagen molecules decreased with the increase of temperature. Moreover, with the increase of pH, the aggregation degree of native and acylated collagen molecules first increased and then decreased gradually. The aggregation degree of native and acylated collagen molecules was the largest near the isoelectric point. Two-dimensional fluorescence spectrum data showed that under the influence of concentration, Phe of native collagen molecules and the excited state of Tyr of acylated collagen molecules responded preferentially. Under the influence of temperature, Tyr of native collagen molecules and the aggregation state of Tyr of acylated collagen molecules responded firstly. Under the influence of pH, Tyr of native collagen molecules and the excited state of Tyr of acylated collagen molecules responded preferentially.
  • QIN Xiuyuan, WEI Ying, LIN Yi, ZHU Yan, GU Ruizeng, PAN Xingchang
    Food and Fermentation Industries. 2022, 48(17): 191-197. https://doi.org/10.13995/j.cnki.11-1802/ts.029957
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    To investigate the promoting effects of collagen peptides (CP) and rice peptides (RP) on skin moisturizing, whitening and anti-oxidation. CP, RP and their complexes on skin care were investigated in vitro, with 5% glycerol as positive control, the water retention of the samples was investigated by weight loss method. Total antioxidant capacity was used to evaluate the antioxidant capacity of samples. The whitening activity of samples was evaluated by tyrosinase inhibition method. Based on the results of in vitro experiments, the proportion of cells was designed. The changes of superoxide dismutase (SOD) activity and reactive oxygen species (ROS) level in mouse melanoma cells (B16) were investigated to reflect the antioxidant capacity of the samples. The activity of tyrosinase in B16 cells was investigated to reflect the whitening ability of B16 cells. The results showed that both CP and RP showed good moisturizing activity in vitro, meanwhile, CP and its complex with RP in a certain proportion were superior to 5% glycerol under the same conditions. Both peptides had antioxidant function in vitro (ORAC method), and could inhibit tyrosinase activity, and synergistic synergistic effect was obtained when they were combined. The complex could scavenge ROS in B16 cells, improve the activity of SOD in B16 cells, and significantly reduce the activity of tyrosinase in B16 cells. The optimal complex ratio was CP:RP=2:1. Moreover, CP showed better antioxidant and moisturizing effects, while RP had better inhibitory effects on tyrosinase activity. The combined raw material could have synergistic effect in promoting moisture retention, anti-oxidation and inhibiting tyrosinase compared to each of them. The combination of collagen peptides and rice peptides in the ratio of 2:1 showed a stronger moisturizing effect than 5% glycerol in vitro, and it also showed a stronger tyrosinase inhibitory activity. In the cell experiment of mouse melanoma cell B16, it could clear the intracellular ROS, improve the activity of intracellular SOD, and significantly reduce the activity of tyrosinase. Collagen peptides and rice peptides at the ratio of 2:1 may play a positive role in skin health improvement, such as moisturizing, whitening and anti-oxidation.
  • HOU Yuke, JIANG Yufei, KANG Mingli, LI Lingyun, HAN Minyi, XU Xinglian
    Food and Fermentation Industries. 2022, 48(17): 198-206. https://doi.org/10.13995/j.cnki.11-1802/ts.029562
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    To efficiently utilize the chicken skeleton and prepare chicken flavor ingredients, the chicken skeleton enzymolysis product was used as raw material. Through the single factor experiment combined with Friedman sorting tests and response surface optimization in combination with sensory evaluation method of fuzzy mathematics, Maillard reaction of enzymolysis liquid from the chicken skeleton in the optimum condition were explored, and the taste and smell before and after the reaction were determined by electronic nose and electronic tongue. The contents of amino acids were measured by HPLC, and volatile substances were estimated by gas chromatography-ion mobility spectroscopy (GC-IMS). The results showed that the optimal conditions of Maillard reaction were pH 6 at 100 ℃ for 60 min. According to the results of electronic nose and electronic tongue, the bitterness of enzymatic hydrolysate was weakened, the unpleasant smells such as amines were reduced, and the overall flavor was enriched after the Maillard reaction. The results of the analysis of amino acid content revealed that the contents of tyrosine, lysine and isoleucine decreased significantly, which indicating that these amino acids were the main amino acids involved in the Maillard reaction. Volatile flavor substances were detected in enzymolysis solution and Maillard reaction products (MRPs), and it was found that alcohols, aldehydes and ketones were the main components of flavor. The present investigation could provide some reference for developing high-quality meat flavoring base material in the industry.
  • TIAN Yue, XU Haoyang, KANG Xiaoou, ZHAO Minzi, RUAN Changqing
    Food and Fermentation Industries. 2022, 48(17): 207-213. https://doi.org/10.13995/j.cnki.11-1802/ts.028558
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    To seek green and safe natural polymer coating film applied to egg preservation, two natural polymer materials, dialdehyde cellulose (DAC) and carboxymethyl cellulose (CMC) were selected to prepare composite films which were applied in coating eggs for fresh-keeping. Firstly, DAC oxidized from cellulose by sodium periodate was heated to make it water-soluble, and then cross-linked with CMC to prepare the composite film (DAC/CMC). The mechanical properties, barrier properties, and hydrophilicity of DAC/CMC were characterized. Furthermore, DAC/CMC was applied as a coating agent for the preservation of the egg, and its effects on the organoleptic quality, weight loss rate, Hastelloy unit, yolk coefficient and pH of the egg white within 60 d were investigated. The results showed that the addition of DAC improved the tensile strength of composite film and further inhibited water vapor and oxygen. It also reduced the hydrophilicity of the composite film. The coating agent had a good inhibitory effect on the reduction of weight, huff unit, yolk coefficient, and pH of egg white. The coating agent with DAC accounting for 12% of the dry weight of CMC and pH 5 was the best condition for fresh-keeping. The polymer composite material has a promising application in green packaging material and the preservation of edible coating film.
  • LIU Wenbo, KE Zhigang, XU Xia, WU Huawei, WANG Qiujuan, ZHOU Xuxia, DING Yuting, LIU Shulai
    Food and Fermentation Industries. 2022, 48(17): 214-221. https://doi.org/10.13995/j.cnki.11-1802/ts.032098
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    In this paper, three different kinds of alginate, alginate + D-sodium isoascorbate, and alginate + compound phosphate were selected as ice glazing solutions for frozen skipjack tuna. Frozen fish was stored at -18 ℃ for 300 d to investigate the effects of different ice glazing solutions on fish freshness quality and oxidation of protein and lipid. The changes in total volatile basic nitrogen (TVB-N), thiobarbituric acid (TBA), peroxide value (POV), fluorescence intensity (FI), weight loss, pH value, drip loss, texture, and sensory characteristics were analyzed during frozen storage. Results showed that the quality of unglazing skipjack tuna decreased significantly after 180 d of frozen storage. During 300 d of frozen storage, ice glazing skipjack tuna significantly inhibited fish corruption, protein and lipid oxidation compared with the control, and also greatly reduced the quality loss of fish. Among them, the combination of D-sodium isoascorbate and alginate had the best effect on freshness quality, lipid oxidation, protein degeneration, and drip loss. After 300 d of storage, the TVB-N value was 7.11% lower than that of the control group, the TBA value was 57.34% lower than that of the control group. Meanwhile, the fluorescence intensity was 21.31% higher than that of the control group and the drip loss rate was 68.18% lower than that of the control group.
  • CAO Siyuan, LI Baoxiang, HE Yue, LIU Min, WU Xiyu, REN Dan, XU Dan
    Food and Fermentation Industries. 2022, 48(17): 222-229. https://doi.org/10.13995/j.cnki.11-1802/ts.028528
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    To investigate the preservation effect of nanocrystalline cellulose (NCC)/chitosan (CS) composite coating on red tangerines (Citrus tangerine Hort. ex Tanaka) at different temperatures, the control fruits and NCC/CS coated fruits were stored at three temperatures of 4, 8 and 12 ℃ in this study. Then the changes in weight loss, decay rate, nutrient content, malondialdehyde content, antioxidant enzyme activity and antioxidant capacity were measured periodically, and the morphologies of the peels and coating films were observed by scanning electron microscopy. The results indicated that there was no significant difference in fruit quality between the control and coated group at 8 ℃. The decay rate, weight loss and malondialdehyde content of the fruit in the coated group at 12 ℃ were lower compared with the control group. Meanwhile, the soluble solids, soluble protein content and peroxidase (POD) activity of the coated group were more stable than that of the control group. Stored at 4 ℃ decreased decay rate in the coated group, maintained antioxidant capacity and increased antioxidant enzyme activity compared with that in the control group. The results of the coating morphology showed that the coating structure on the fruit skin stored at 4 ℃ was the most stable and the microporosity appeared at the latest. In general, NCC/CS coating significantly delayed the senescence of red tangerines fruits and improved their storage quality at 12 ℃, however, it enhanced the tolerance of fruit to chilling injury at 4 ℃. Therefore, coating treatment could effectively improve the storage quality of red tangerines under unsuitable temperatures and reduce the decay rate caused by temperature fluctuations during storage and transportation.
  • GUO Jie, SHI Feng, SUN Manman, MA Fanqi, LI Yongfu
    Food and Fermentation Industries. 2022, 48(17): 230-235. https://doi.org/10.13995/j.cnki.11-1802/ts.032096
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    Sodium nitrite is one of the most toxic food additives, and insoluble dietary fiber (IDF) can scavenge sodium nitrite. To reduce the harm caused by sodium nitrite to humans, this paper studied the chemical composition of Eucheuma sp. waste residue after extracting carrageen. Then the Eucheuma sp. waste residue was pulverized to be ultrafine powder, and their hydration characteristics and cation exchange capacity were determined. On this basis, their capacity to adsorb sodium nitrite and ability to block nitrosamine synthesis under the simulated gastric juice in vitro was investigated. Results showed that the IDF content of Eucheuma sp. waste residue was up to 90.1%. Their maximum water holding capacity and expansion capacity reached (5.61±0.06) g/g and (5.67±0.30) mL/g, respectively, and their maximum cation exchange capacity was (0.32±0.02) mmol NaOH/g. More interestingly, they could adsorb (70.0±0.4)% sodium nitrite within 5 min and the adsorption ratio reached (93.1±0.7)% after equilibrium. The IC50 value was 0.87 mg/mL, and the adsorption capacity was up to (18.376±0.066) mg/g. The adsorption accorded with the Freundlich isotherm model and pseudo-second-order kinetic model. Furthermore, their blocking rate of nitrosamine synthesis reached (65.2±2.1) %. In conclusion, the ultrafine powder of Eucheuma sp. waste residue has excellent physicochemical properties, sodium nitrite adsorption capacity, and nitrosamine synthesis blocking ability, thereby it is suitable for developing sodium nitrite scavengers.
  • FAN Ying, DU Lin, HAN Jiamin, CHEN Qingju, ZHANG Li, HOU Ping
    Food and Fermentation Industries. 2022, 48(17): 236-240. https://doi.org/10.13995/j.cnki.11-1802/ts.031310
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    In this study, the shallow empty honeybee combs were used to produce mature honey. The combs with completely cover honey was taken out from the hive and put into the drying chamber (temperature 26-28 ℃, humidity 18%-20%) for post-ripening treatment. The effects of different post-ripening treatment time (0, 24, 72, 120, 168 h) on the water content, sugar, conductivity, 5-hydroxymethyl furfural, amylase and other components in gallnut honey were analyzed. The results showed that, with the extension of post-ripening treatment time, the water content and amylase in gallnut honey decreased gradually, while electrical conductivity, sucrose and 5-hydroxymethyl furfural increased. But glucose, fructose and chromaticity did not change significantly. After post-ripening treatment for 120 h, water content, conductivity, sucrose, glucose, fructose, 5-hydroxymethyl furfural and amylase value in gallnut honey decreased to 19.63%, 467.93 μS/cm, 3.32%, 31.45%, 35.75%, 1.03 mg/kg and 11.31 mL/(g·h) respectively, which have met the quality standard of first-grade honey in China. The post-ripening treatment method avoided the destruction of nutritional components under high temperature concentration. This study could be used to improve the processing technology of honey products and guide the mechanization of beekeeping industry.
  • LING Yuting, CHEN Shuang, XU Yan, FAN Wenlai
    Food and Fermentation Industries. 2022, 48(17): 241-248. https://doi.org/10.13995/j.cnki.11-1802/ts.031416
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    The volatile compounds in Gujinggong Baijiu with different aging times were characterized by headspace solid phase microextraction(HS-SPME) with comprehensive two-dimensional gas chromatography/time-of-flight mass spectrometry(GC×GC-TOFMS). Through the identification strategy, 262 potential aroma active components were accurately identified. In addition to alcohol, aldehyde, acid, and ester compounds, a variety of trace aroma compounds in Gujinggong Baijiu were richest. 50 substances with significant differences in the content of Gujinggong Baijiu at different aging times were analyzed by multivariate statistical analysis method. Most of the furan, terpene, aromatic and acid flavor compounds gradually increased with aging time, which contributed to the aroma of Baijiu samples. Chemometrics methods were used to analyze the Baijiu samples with different grades and aging times. The results showed that the differences of grades among Baijiu samples were more important than the differences of years, which confirmed that the quality control of Baijiu had certain significance for regulating the flavor quality of Baijiu aging.
  • LIU Huijie, WANG Juan, WANG Zhongchao, HUANG Mingquan, LIU Yongguo, WU Jihong, ZHANG Jinglin
    Food and Fermentation Industries. 2022, 48(17): 249-255. https://doi.org/10.13995/j.cnki.11-1802/ts.030718
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    The volatile compounds in Fangxian Huangjiu (FXHJ) were analyzed by sensory evaluation, headspace solid phase microextraction (HS-SPME) combined with two-dimensional gas chromatography-mass spectrometry (GC×GC-MS). Three samples, including Fuzhi (fermented for 3 d), dry-type and sweet-type Huangjiu were investigated. The sensory evaluation results showed that the main aroma profiles of FXHJ were rice aroma, alcoholic, floral, fruity, sweet, acidic, Zao-aroma (fermented grain aroma) and roast aroma, and the characteristic attributes of FXHJ were the strong aroma of rice, fruity and floral. A total of 205 volatile compounds were detected in FXHJ. The total content of volatiles in Fuzhi (fermented for 3 d) was the highest 20 867.26 μg/L, followed by dry-type 16 020.60 μg/L, and sweet-type Huangjiu had the lowest concentrations 14 287.75 μg/L. Partial least squares regression analysis predicted the β-phenylethanol, 3-methylthio-1-propanol, ethyl propionate, 3-methylbutanal, 1-octen-3-ol, 4-ethylphenol, ethyl acetate, nonanal, 3-methyl-1-butanol, 1-propanol, 3-methylbutanoic acid, 2-methyl-1-propanol and phenylethyl acetate significantly positively associated with the rice and sweet aroma, while diethyl succinate, ethyl lactate, isoamyl acetate, 4-octanone, and ethyl pentadecanoate significantly associated with the acidic, fruit, Zao-aroma and alcoholic notes.
  • ZHANG Chen, ZHOU Jia, MA Yaqin, DENG Tujing, WANG Jun
    Food and Fermentation Industries. 2022, 48(17): 256-264. https://doi.org/10.13995/j.cnki.11-1802/ts.028946
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    The aim of this study was to compare the specific differences in the composition and content of free and bound aroma substances in wide peel citrus (Nanfeng, Ponkan), oranges (Qichen, Changyexiangchen), hybrid citrus (Aiyuan 38, Wogan) and Guanxi Honey Pomelo, in order to provide a scientific basis for the flavor characteristics of citrus fruit juice. Headspace solid-phase microextraction gas chromatography-mass spectrometry (HS-SPME-GC-MS) was used to extract the volatile components from the four citrus juices, and principal component analysis (PCA) was used to distinguish them. A total of 127 bound and free aroma substances were characterized in the four categories of orange juice, among which olefins yields were the highest (50); alcohols, aldehydes and esters were the second (20, 19 and 17 respectively); ketones and phenols were less (6 and 5 respectively). In terms of content, the olefins yield in the total aroma substances from the total orange juice samples were more than 90%, followed by alcohols, aldehydes, esters, ketones and phenols, respectively. In addition, the four categories of citrus juice were taken as the research object, and the target aroma substances were analyzed by principal component analysis. The four categories of citrus juice could be distinguished by PCA analysis. The content of free aroma substances in hybrid citrus was much lower than wide peel citrus and orange (their content was similar), and much higher than guanxi pomelo. However, the free aroma substances contained in hybrid citrus were rich, and among which Aiyuan 38 was the most abundant, and was much higher than other citrus varieties (the number of free aroma substances contained in other citrus varieties was similar). In addition, broad peel citrus contained the most kinds of bonded aroma compounds, followed by orange, while Guanxi pomelo was the least.
  • LIU Yao, QIAO Haijun, JIA Zhilong, ZHANG Weibing, ZHANG Chunyan, HUO Shanglei, MIAO Jingyuan, SHEN Mengna
    Food and Fermentation Industries. 2022, 48(17): 265-272. https://doi.org/10.13995/j.cnki.11-1802/ts.029646
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    The changes of volatile flavor compounds in yak milk cheese during ripening were analyzed by gas chromatography-ion mobility spectrometry. The results showed that 22 volatile flavor compounds from 46 signal peaks were identified. The contents of volatile flavor compounds of yak milk cheese at different ripening time were significantly different. The contents of acetoin and ethanol were highest at the early ripening stage (1 d); the content of 2,4,6-trimethylpyridine, isoamyl acetate and ethyl butyrate increased gradually during the middle stage of ripening (30, 60 and 90 d); At the late stage of ripening (120, 150 and 180 d), the compounds including ethyl butyrate, ethyl hexanoate dimer, isoamyl acetate and ethyl acetate dimer increased to maximum value. Twelve potential characteristic volatile markers (VIP>1) were screened out by partial least squares-discriminant analysis, including 3-methylbut-3-en-1-ol, ethyl acetate dimer, acetoin, isoamyl acetate, ethanol, ethyl butyrate, 2-butanone, n-butanol, ethyl hexanoate dimer, allyl nitrile, 2-pentanone dimer and benzaldehyde were screened out. The results of principal component analysis can facilely distinguish the ripening stages of yak milk cheese. This study provides a reference for the development and quality control of yak milk cheese products in future.
  • ZHANG Mingjuan, LIU Mingming, YU Qiudi, YUAN Lei, WANG Juan, QIN Ai, LI Genrong
    Food and Fermentation Industries. 2022, 48(17): 273-281. https://doi.org/10.13995/j.cnki.11-1802/ts.031352
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    To establish a multiplex PCR method for the simultaneous detection of 10 kinds of foodborne pathogens in food, including Listeria monocytogenes, Salmonella spp., Cronobacteria spp., Shigella spp., Staphylococcus aureus, Vibrio parahaemolyticus, Escherichia coli O157: H7, Pseudomonas aeruginosa, Streptococcus faecalis and Clostridium perfringens, the multiplex PCR detection system was established by specific primer design, primer specificity verification, primer sensitivity verification and optimization of the main reaction conditions. Furthermore, the specificity, sensitivity and application of artificially infected samples of the multiplex PCR system were evaluated. The results showed that the specific target bands of 10 foodborne pathogens could be amplified by the multiplex PCR system without nonspecific amplification. The sequencing results showed 98% homology with the target gene sequence, and the sensitivity of the system was up to 10-1 ng/μL. The application results of artificially contaminated samples showed that the multiplex PCR method was simple and rapid, requiring only simple multiplication, with a detection limit of up to 10 CFU/mL, and the detection time was less than 48 h. The multiplex PCR detection system have strong specificity and high sensitivity, which is closely related to actual detection work of inspection institutions, and have great promotion and application value.
  • HUANG Hua, XIE Wendong, GU Yu, MENG Zhaoqi, SUN Yuanjie, DU Hongmiao, SUN Yana, KANG Shuai, LI Mengyun
    Food and Fermentation Industries. 2022, 48(17): 282-290. https://doi.org/10.13995/j.cnki.11-1802/ts.029524
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    This study intended to develop a method for the determination of 50 pesticide residues in fruits and vegetables by sample purification with QuEChERS of combing multi-walled carbon nanotube and ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Because of its unique hollow structure and large specific surface area, multi-walled carbon nanotube has excellent adsorption properties, which make it shows superior purification efficiency in the sample pre-treatment process of pesticide residue analysis of fruits and vegetables. This paper selected and determined the type and usage of multi-walled carbon nanotubes through optimization experiments. Based on the QuEChERS method, this paper improved the sample purification effect. This paper extracted fruit and vegetable samples with 0.1% formic acid acetonitrile solution. Meanwhile, this paper detected 50 pesticide residues in fruits and vegetables through ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). The results demonstrated that they were 0.002-0.010mg/kg for the limits of quantification (LOQs) for 50 compounds, 71%-105% recoveries of low, medium and high supplemental levels within the linear range and 3.1%-11.2% relative standard deviation (RSD). Good linearity of the Matrix matching standard curve was observed for all calibration curves with correlation coefficients (r) > 0.99. This method was rapid, high sensitivity and accuracy. It is suitable for rapid detection and analysis of pesticide residues in fruit and vegetable samples.
  • WANG Huifang, YU Yongxin, LI Chenhong, CHEN Wenjun, LING Lanxin, ZHOU Ying
    Food and Fermentation Industries. 2022, 48(17): 291-297. https://doi.org/10.13995/j.cnki.11-1802/ts.031606
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    In this study, a method of recombinase polymerase amplification (RPA) combined with lateral flow dipstick (LFD) technology (RPA-LFD) was established for the characterization of Atlantic cod related products. Based on the comparison between the sequences of mitochondrial genomes of the Atlantic cod and popular adulterated cod on the market, specific probes and primers were designed for detecting the Atlantic cod and an RPA-LFD detection method for the Atlantic cod were established. The objectives of this study were to select the best primers and probes, evaluate their specificity and optimize the best reaction conditions, which including temperature, reaction time both in the lab and in the field. The optimized method was used to perform the real-time detection of 20 ‘cod' products and common adulterated ones that were currently available on the market. The results demonstrated that RPA-LFD showed a strong specificity by incubating the reaction at 44 ℃ for 14 min. The results of real-time detection of 20 ‘cod' products were consistent with the true identity of the product, which was confirmed by sequencing results. The method of RPA-LFD for distinguishing Atlantic cod and related products established in this study showed a strong specificity and with the advantages of short reaction time, simple protocol, and visible results. It provides a new strategy for real-time identification of cod products in the field, which is important for the supervision of the market of aquatic products.
  • LU Yinyue, SONG Xin, XIA Yongjun, WANG Guangqiang, AI Lianzhong, XIONG Zhiqiang
    Food and Fermentation Industries. 2022, 48(17): 298-303. https://doi.org/10.13995/j.cnki.11-1802/ts.030769
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    Yogurt starter is a microbial material composed of lactic acid bacteria (LAB), mainly Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus. The two-component signal transduction system (TCS) composed of the transmembrane receptor histidine protein kinase and the homologous intracytoplasmic response regulator protein is a common signal mechanism in bacteria, which enables bacteria to sense external stimuli and regulate their own physiological and biochemical processes. Compared with the model bacteria such as Escherichia coli, Bacillus subtilis, and Streptomyces coelicolor, the response and regulation mechanism of LAB TCS is lack of systematic elucidation. This article summarizes the progress in the TCS of L. delbrueckii subsp. bulgaricus and S. thermophilus, with a view to provide help for the research of LAB TCSs and the industrial production control of yogurt.
  • YANG Qinghua, ZHANG Yafei, TIAN Xiaojing, WANG Wenhang
    Food and Fermentation Industries. 2022, 48(17): 304-312. https://doi.org/10.13995/j.cnki.11-1802/ts.031278
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    Fermentation is one of the most economical and safe methods to improve the nutritional value, sensory quality and functional characteristics of raw materials, and it is also an important method to process grain. This paper summarizes the latest research progress of grain fermentation products and related biotransformation. It is focused on the improvement of nutritional value and prebiotic potential, including the effect of fermentation on the digestibility of grain protein and starch, the reduction of anti-nutrients and the enhancement of phenolic compounds and vitamins. As a prebiotic functional substance, cereal fermented products have great potential to maintain the balance of intestinal flora, and also have a positive effect on cardiovascular system and gastrointestinal tract. Finally, the activity function evaluation and future market development of fermented cereal products have been summarized.
  • SHANGGUAN Lingling, LU Huifang, XIA Huili, CHEN Xiong, DAI Jun
    Food and Fermentation Industries. 2022, 48(17): 313-320. https://doi.org/10.13995/j.cnki.11-1802/ts.030792
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    Corynebacterium glutamicum, as a core strain for industrial production of amino acids, has been developed into a multifunctional microbial cell factory that can use renewable raw materials to produce a variety of valuable natural or unnatural chemicals. Aiming at the new construction technology and application fields of C. glutamicum cell factory in recent years, this review first summarizes the new mutation, high-throughput screening and gene editing technology for the construction of C. glutamicum cell factory, and then introduces the application of the engineered microbial cell factory in the efficient synthesis of organic acids, biofuels and terpenes. Finally, some classic examples of efficient microbial cell factory using renewable raw materials to produce high value-added products are listed. In summary, it provides a reference for expanding the scope of C. glutamicum product and substrate.
  • ZHANG Xinran, LING Yan, YANG Ying
    Food and Fermentation Industries. 2022, 48(17): 321-328. https://doi.org/10.13995/j.cnki.11-1802/ts.029940
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    Pichia pastoris expression system has many excellent characteristics and is one of the most successful foreign protein expression systems. Through the rapid development in recent decades, it has attracted wide attention from all walks of life, and has produced huge economic and social value. Compared with other existing expression systems, P. pastoris has great advantages in post-translational modification of target proteins, and has been widely used to express exogenous proteins. However, on the other hand, the system is affected by many factors, including the characteristics of exogenous genes, bacterial species, expression environment and fermentation technology, so it also shows great differences in the expression of different exogenous proteins. Gene transcription module, this paper focuses on the molecular level of protein secretion and degradation of modules and transport four modules, and introduces the use of P. pastoris expression system by optimizing codon, high copy number exogenous gene screening, gene knockout protease, and guide the peptides were folded factors to improve the efficiency of heterologous protein expression of related policies, The development prospect of the system is briefly prospected.
  • XIE Yuejie, WANG Kai, XIAO Yixue, WANG Cao, WANG Yihan, WANG Qiang
    Food and Fermentation Industries. 2022, 48(17): 329-335. https://doi.org/10.13995/j.cnki.11-1802/ts.031913
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    Nanoemulsion has many potential advantages over traditional emulsion, especially in special foods such as baby food, functional beverage and so on. Long chain polyunsaturated fatty acids, phytosterols, carotenoids and other liposoluble substances usually need to be embedded before they can be evenly dispersed in aqueous food. However, because of the larger size of carrier particles and the carrying of active substances, ordinary emulsion cannot show clarified, transparent, uniform and other quality characteristics, which also limits the transmission system of liposoluble bioactive substances. In this paper, the preparation methods, emulsion characteristics and bioavailability of long chain n-3 polyunsaturated fatty acid nanoemulsion were reviewed in order to provide references for the preparation and application of nano emulsion with functional components.
  • ZOU Yuanting, ZHANG Jian, WANG Gongming, JING Yuexin, LIU Fang, JIAO Chunna, ZHONG Jingshi, WANG Yixin
    Food and Fermentation Industries. 2022, 48(17): 336-344. https://doi.org/10.13995/j.cnki.11-1802/ts.029941
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    Bioactive peptides(BAPs) are a short sequence of peptides composed of about 2-20 amino acids that are beneficial to biological health, as molecular polymers between amino acids and proteins, BAPs have beneficial effects such as anti-hypertension, anti-oxidation, anti-cancer, anti-inflammation, anti-bacterial, and promoting wound healing. It also has the functions of immune regulation and biological regulation. In recent years, bioactive peptides have been widely used in formula foods, functional foods and biomedical fields due to their unique physical and chemical properties, health promotion, disease prevention or delay and high safety. Based on the physical and chemical properties and characteristics of bioactive peptides, this review focuses on the research progress of nutritional interventions of bioactive peptides in patients with gastrointestinal dysfunction, cancer, diabetes and muscle attenuation. The potential applications and future development of BAPs are also discussed to provide certain reference values to the applications and researches of bioactive peptides in FSMPs.
  • ZHANG Yue, LI An, ZHAO Jie, PAN Ligang
    Food and Fermentation Industries. 2022, 48(17): 345-352. https://doi.org/10.13995/j.cnki.11-1802/ts.029749
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    Postharvest vegetables are fresh and perishable farm produces. However, due to the strong physiological metabolism, pathogenic microorganism infection and incorrect storage methods, they are very prone to decay and cause a heavy loss therefore. While, properly using the preservatives not only could maintain the quality of the vegetables, but also facilitates their storage and transportation. Over recent years, preservatives have played an increasingly important role in the preservation, storage and circulation of post-harvest vegetables. Starting with a brief description of the principles of preservatives, this paper classified the preservatives according to their mechanisms and characteristics and further explained from four aspects: biological preservatives, chemical preservatives, plant growth substance preservatives and food additive preservatives. Research progress in the use of botanical, animal, microbial and chemical sourced preservatives in vegetable storage and transportation was also carefully elaborated in this article. Finally, a prospect of researching and developing high-efficient, safe and healthy preservative agents, as well as the compound preservative agent was proposed at the end of this paper.
  • WANG Jinjing, ZHENG Feiyun, NIU Chengtuo, LIU Chunfeng, LI Qi
    Food and Fermentation Industries. 2022, 48(17): 353-356. https://doi.org/10.13995/j.cnki.11-1802/ts.031610
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    Brewing industry is an important part of China's food and beverage industry, and brewing masters' training is the main task for brewing talents' cultivation. Brewing Process Experimental Course is an important practical course for students majoring in brewing engineering, which requires students to combine theoretical knowledge with brewing practice and improve their engineering practice ability. Based on the phenomenon of teacher-oriented teaching in current teaching methods, Project-Based Experimental Teaching (PBET) method was applied to the Brewing Process Experimental Course in this study, trying to explore the role of experiential learning method in students' engineering practice learning process. PBET helps improve students' autonomous learning ability. In the process of completing the assigned project, students can constantly revise and improve the project proposal, finish the experimental practice, through effective online/offline communication with teachers, and further cultivates students' engineering practical ability. When applying this method to the experimental course of brewing process, students complete the design of beer recipe, the brewing test and finally submit the final product via the whole cycle learning of Do-Reflect-Think-Apply. It is shown to receive good teaching effects through PBET method.