Food and Fermentation Industries >

2023 , Vol. 49 >Issue 17: 37 - 43

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.033847

Isolation and purification of a cellulose-producing strain and its enzyme-producing properties

  • HUANG Cuixin ,
  • ZHANG Guirong ,
  • LIU Jun ,
  • SHU Lin ,
  • CAO Rong ,
  • WEN Xueping ,
  • LI Li
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  • 1(College of Bioengineering, Sichuan University of Science & Engineering, Yibin 644000, China)
    2(Sichuan Slan Biotechnology Co.Ltd., Chengdu 610000, China)
    3(Sichuan Taiyuanjing Vinegar Co.Ltd., Zigong 643000, China)
    4(Soild-state Fermentation Resource Utilization Key Laboratory of Sichuan Province, Yibin 644000, China)

Received date: 2022-10-04

  Revised date: 2022-11-05

  Online published: 2023-09-27

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Abstract

In this research, a fungus with high cellulase activity was screened from Cupei of Sichuan sun vinegar. Its physiological-biochemical characteristics were studied and analyzed by ITS sequence. The conditions of enzyme producing and enzymatic properties of the strain were investigated by single factor and orthogonal experiments. The strain was identified as Lichtheimia ramosa, the crude enzyme solution was extracted under the following conditions: 3.5×106 CFU/mL of bacterial suspension, 1% of inoculation volume, 30 ℃, cultured for 24 h at 180 r/min, and the reaction was performed at 50 ℃ for 30 min. The max activities of endoglucanase and β-glucosidase were 95.92 U/mL and 47.57 U/mL at pH 5.0 and 6.0. This study revealed that endoglucanase and β-glucosidase were produced by L. ramosa during the fermentation of Cupei, and had well stability of pH and heat stress, which provided some reference value for the production of Sichuan sun vinegar.

Key words： Sichuan sun vinegar; isolation and identification; Lichtheimia ramosa; endoglucanase; β-glucosidase

Cite this article

HUANG Cuixin , ZHANG Guirong , LIU Jun , SHU Lin , CAO Rong , WEN Xueping , LI Li . Isolation and purification of a cellulose-producing strain and its enzyme-producing properties[J]. Food and Fermentation Industries, 2023 , 49(17) : 37 -43 . DOI: 10.13995/j.cnki.11-1802/ts.033847

References

[1] 朱文优, 张超, 魏琴, 等.四川晒醋烘醅工艺的改良研究[J].中国调味品, 2012, 37(4):82-84;91.
ZHU W Y, ZHANG C, WEI Q, et al.Technical innovation in the baking fermented grains processing of Sichuan Sun vinegar[J].China Condiment, 2012, 37(4):82-84;91.
[2] 周荣清. 四川麸醋生产工艺的特点及发展趋势[J].四川食品与发酵, 2000, 36(3):24-28.
ZHOU R Q.Characteristics and development trend of Sichuan bran vinegar production technology[J].Sichuan Food and Fermetation, 2000, 36(3):24-28.
[3] 张桂容, 冯洁雅, 蔡吉, 等.四川晒醋固态发酵过程中理化因子与真菌群落结构的动态变化规律[J].食品工业科技, 2022, 43(9):131-138.
ZHANG G R, FENG J Y, CAI J, et al.Dynamic changes of physicochemical properties and fungal community structure during solid-state fermentation of Sichuan Sun vinegar[J].Science and Technology of Food Industry, 2022, 43(9):131-138.
[4] 冯洁雅, 张桂容, 蔡吉, 等.基于高通量测序解析四川晒醋固态发酵过程中细菌群落变化[J].食品工业科技, 2021, 42(19):135-143.
FENG J Y, ZHANG G R, CAI J, et al.Analysis of bacterial community changes in Sichuan Sun vinegar solid-state fermentation process based on high-throughput sequencing[J].Science and Technology of Food Industry, 2021, 42(19):135-143.
[5] 鲍晓华, 董维多.纤维素酶在食品发酵和加工中的应用[J].思茅师范高等专科学校学报, 2010, 26(3):26-29.
BAO X H, DONG W D.Application of cellulase in food fermentation and processing[J].Journal of Simao Teacher’s College, 2010, 26(3):26-29.
[6] 邓天福, 杜开书, 李广领.纤维素酶及其在酿造业中的应用[J].中国酿造, 2011, 30(12):17-19.
DENG T F, DU K S, LI G L.Cellulase and its application in brewing industry[J].China Brewing, 2011, 30(12):17-19.
[7] ZANG X Y, LIU M T, WANG H, et al.The distribution of active β-glucosidase-producing microbial communities in composting[J].Canadian Journal of Microbiology, 2017, 63(12):998-1008.
[8] 阎伯旭, 齐飞, 张颖舒, 等.纤维素酶分子结构和功能研究进展[J].生物化学与生物物理进展, 1999, 26(3):233-237.
YAN B X, QI F, ZHANG Y S, et al.Progress in structure function studies of cellulases[J].Progress in Biochemistry and Biophysics, 1999, 26(3):233-237.
[9] TAO P, ZHANG Y H, WU Z M, et al.Enzymatic pretreatment for cellulose nanofibrils isolation from bagasse pulp:Transition of cellulose crystal structure[J].Carbohydrate Polymers, 2019, 214:1-7.
[10] 苑广伟, 周洁嫦, 王欣宇, 等.一株来源于土壤的横梗霉菌及其产羧甲基纤维素酶性质[J].工业微生物, 2021, 51(1):23-29.
YUAN G W, ZHOU J C, WANG X Y, et al.A Sporidiobolus pararoseus XWNR1 from soil and its properties of carboxymethyl cellulase[J].Industrial Microbiology, 2021, 51(1):23-29.
[11] 刘军, 江若宇, 王麒麟, 等.四川晒醋醋醅中一株产纤维素酶的芽孢杆菌分离筛选[J].食品与发酵工业, 2018, 44(6):140-144.
LIU J, JIANG R Y, WANG Q L, et al.Screening of cellulose-producing Bacillus from Sichuan Sun vinegar[J].Food and Fermentation Industries, 2018, 44(6):140-144.
[12] 吴佳. 链霉菌AJ56的生理生化特性研究[J].长治医学院学报, 2022, 36(1):12-14.
WU J.Study of the physiological and biochemical characteristics of Streptomyces AJ56[J].Journal of Changzhi Medical College, 2022, 36(1):12-14.
[13] 马鹏. 高酯化力、糖化力和液化力霉菌的筛选及强化大曲的研究[D].阿拉尔:塔里木大学, 2022.
MA P.Screening of molds with high esterification, saccharification and liquefaction ability and study on strengthening Daqu[D].Ala’er:Tarim University, 2022.
[14] WILLIAMS S T, SHARPE E M, HOLT J G.Bergey’s Manual of Systematic Bacteriology[M].Philadelphia:The William’s & Wilkins Company, 1984, 23:103-107.
[15] 张纪忠. 微生物分类学[M].上海:复旦大学出版社, 1990:317-325.
ZHANG J Z.Microbial Taxonomy[M].Shanghai:Fudan University Press, 1990:317-325.
[16] 邓诗贵. 白酒糟粗纤维降解真菌的筛选与降解工艺的初步研究[D].南昌:江西农业大学, 2018.
DENG S G.Screening of fungi degrading crude fiber from distiller’s grains and preliminary study on degradation technology[D].Nanchang:Jiangxi Agricultural University, 2018.
[17] 郭金玲, 陈程鹏, 周一郎, 等.黑曲霉β-葡萄糖苷酶的分离纯化及酶学性质研究[J].中国酿造, 2021, 40(2):83-87.
GUO J L, CHEN C P, ZHOU Y L, et al.Isolation, purification and enzymatic properties of β-glucosidase from Aspergillus niger[J].China Brewing, 2021, 40(2):83-87.
[18] 江学斌, 成雪鸿, 马苗鹏, 等.产纤维素酶芽孢杆菌的鉴定与酶活力比较[J].广东农业科学, 2022, 49(4):116-122.
JIANG X B, CHENG X H, MA M P, et al.Identification and enzyme activity comparison of cellulase-producing Bacillus[J].Guangdong Agricultural Sciences, 2022, 49(4):116-122.
[19] S.A.瓦克斯曼著〔美〕,阎逊初译.放线菌属和种分类、鉴定和描述(第二卷)[M]. 北京:科学出版社, 1974:187-188.
S.A.WAKSMAN[USA], translated by YAN X C.Taxonomy, Identification and Description of Genera and Species of Actinomycetes (vol.2)[M].Beijing: Science Press, 1974:187-188.
[20] 魏景超. 真菌鉴定手册[M].上海:上海科学出版社, 1979.
WEI J C.Manual of Fungal Identification[M].Shanghai:Shanghai Science Press, 1979.
[21] 白茹玉, 石彥鹏, 张萍.不同培养条件对林可链霉菌种子液代谢影响[J].中国兽药杂志, 2022, 56(3):53-59.
BAI R Y, SHI Y P, ZHANG P.Effects of different culture conditions on metabolism of seed fluid of Streptomyces lincolnensis[J].Chinese Journal of Veterinary Drug, 2022, 56(3):53-59.
[22] 雅男. 纤维素降解菌的筛选及酶学性质的研究[D].哈尔滨:东北林业大学, 2017.
YA N.Screening of cellulose degrading bacteria and studies on cellulase characteristics[D].Harbin:Northeast Forestry University, 2017.
[23] HONG S B, KIM D H, LEE M N, et al.Zygomycota associated with traditional meju, a fermented soybean starting material for soy sauce and soybean paste[J].Journal of Microbiology, 2012, 50(3):386-393.
[24] 孙启星, 曾小波, 李学伟, 等.酱油曲中谷氨酰胺酶酶学特性研究[J].中国调味品, 2021, 46(4):82-85.
SUN Q X, ZENG X B, LI X W, et al.Research on the enzymatic properties of glutaminase in soy sauce koji[J].China Condiment, 2021, 46(4):82-85.
[25] 徐菲, 朱慧霞, 王淮, 等.黄曲霉YZC产纤维素酶的固态发酵条件优化[J].纤维素科学与技术, 2014, 22(4):44-51.
XU F, ZHU H X, WANG H, et al.Optimization of solid-state fermentation conditions for cellulase production with Aspergillus flavus YZC[J].Journal of Cellulose Science and Technology, 2014, 22(4):44-51.
[26] XIE J H, PAN L, HE Z M, et al.A novel thermophilic β-mannanase with broad-range pH stability from Lichtheimia ramosa and its synergistic effect with α-galactosidase on hydrolyzing palm kernel meal[J].Process Biochemistry, 2020, 88:51-59.
[27] DE OLIVEIRA A P A, SILVESTRE M A, GARCIA N F L, et al.Production and catalytic properties of amylases from Lichtheimia ramosa and Thermoascus aurantiacus by solid-state fermentation[J].The Scientific World Journal, 2016, 2016:1-10.
[28] YANG S, LEE J, KWAK J, et al.Fungi associated with the traditional starter cultures used for rice wine in Korea[J].Journal of the Korean Society for Applied Biological Chemistry, 2011, 54(6):933-943.
[29] YANG J G, DOU X, HAN P J, et al.Microbial diversity in daqu during production of Luzhou flavored liquor[J].Journal of the American Society of Brewing Chemists, 2017, 75(2):136-144.
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