前期初探发现枯草芽孢杆菌(Bacillus subtilis) BJ3-2发酵精薏米可高产川芎嗪[2.97 mg/g 干基(dry weight,DW)]和纤溶酶(720.84 U/g)。该研究以糙薏米、精薏米和碎薏米为原料,以B.subtilis BJ3-2为发酵菌株,通过单因素及Box-Behnken试验构建高产川芎嗪和溶纤酶的薏米发酵体系。结果表明,接种量为9.0%、发酵温度为40 ℃和发酵时间为93 h时,B.subtilis BJ3-2发酵糙薏米中川芎嗪产量为6.15 mg/g DW、溶纤酶酶活为2 236.47 U/g,而以接种量为8.1%、发酵温度为38 ℃和发酵时间为96 h时,发酵精薏米中川芎嗪产量为6.94 mg/g DW、溶纤酶酶活为2 142.18 U/g,但碎薏米中川芎嗪产量和溶纤酶酶活均明显偏低。此外,B.subtilis BJ3-2分别发酵的3种薏米,其川芎嗪含量均高于B.subtilis BJ3-2发酵黄豆和CICC 20637发酵薏米。因此,综合考虑加工成本,B.subtilis BJ3-2发酵糙薏米可作为高产川芎嗪和溶纤酶的最佳发酵体系。
Our previous study found that high-yield tetramethylpyrazine (2.97 mg/g DW) and fibrinolytic enzyme (720.84 U/g) were obtained in Bacillus subtilis BJ3-2-fermented polished adlay. Taking dehulled, polished and broken adlay as raw materials and B. subtilis BJ3-2 as fermentation strain, the fermentation conditions for high-yield tetramethylpyrazine and fibrinolytic enzyme production were optimized by single factor and Box-Behnken experiments in this study. Results showed that the tetramethylpyrazine yield and fibrinolytic enzyme activity of B. subtilis BJ3-2-fermented dehulled adlay were 6.15 mg/g DW and 2236.47 U/g when inoculation amount, fermentation temperature and fermentation time were 9.0%, 40 ℃ and 93 h. The tetramethylpyrazine yield and fibrinolytic enzyme activity of B. subtilis BJ3-2-fermented polished adlay were 6.94 mg/g DW and 2142.18 U/g when inoculation amount, fermentation temperature and fermentation time were 8.1%, 38 ℃ and 96 h. The tetramethylpyrazine yield and fibrinolytic enzyme activity in B. subtilis BJ3-2-fermented broken adlay were low even after optimization. Additionally, the tetramethylpyrazine yield of B. subtilis BJ3-2-fermented adlay was higher than that of B. subtilis BJ3-2-fermented soybean and CICC 20637-fermented adlay. Therefore, considering the processing cost of adlay, B. subtilis-fermented dehulled adlay could be the optimal fermentation system to obtain high-yield tetramethylpyrazine and fibrinolytic enzyme.
[1] YE J, HUANG L, TEREFE N S, et al.Fermentation-based biotransformation of glucosinolates, phenolics and sugars in retorted broccoli puree by lactic acid bacteria[J].Food Chemistry, 2019, 286:616-623.
[2] DING Y, PU L, KAN J.Hypolipidemic effects of lipid-lowering granulated tea preparation from monascus-fermented grains (adlay and barley bran) mixed with lotus leaves on Sprague-Dawley rats fed a high-fat diet[J].Journal of Functional Foods, 2017, 32:80-89.
[3] LI P, TSAI W, CHIEN C.Dietary monascus adlay supplements facilitate suppression of cigarette smoke-induced pulmonary endoplasmic reticulum stress, autophagy, apoptosis and emphysema-related PLGF in the rat[J].Food Chemistry, 2013, 136(2):765-774.
[4] YIN H M, ZHONG Y D, XIA S K, et al.Effects of fermentation with Lactobacillus plantarum NCU137 on nutritional, sensory and stability properties of Coix (Coix lachryma-jobi L.) seed[J].Food Chemistry, 2020,134:126 037.
[5] WANG C, LIN H, WU S.Influence of dietary supplementation with Bacillus-fermented adlay on lipid metabolism, antioxidant status and intestinal microflora in hamsters[J].Journal of the Science of Food and Agriculture, 2011, 91(12):2 271-2 276.
[6] WEN A, XIE C, MAZHAR M, et al.Comparative evaluation of drying methods on kinetics, biocompounds and antioxidant activity of Bacillus subtilis-fermented dehulled adlay[J].Drying Technology, 2020, 38(11):1 505-1 515.
[7] WEN A, QIN L, ZENG H, et al.Comprehensive evaluation of physicochemical properties and antioxidant activity of B.subtilis-fermented polished adlay subjected to different drying methods[J].Food Science & Nutrition, 2020, 8(4):2 124-2 133.
[8] 涂鸿.贵州薏米原料及加工产品有害重金属污染剖析[D].贵阳:贵州大学, 2016.
TU H.Analysis of the harmful heavy metal pollution in Coix Rice raw material and processing products from Guizhou[D].Guiyang:Guizhou University, 2016.
[9] 遇靓.红曲固态发酵薏米茶工艺及品质研究[D].贵阳:贵州大学, 2016.
YU L.Study on processing technology and quality of solid fermented tea by monacus[D].Guiyang:Guizhou University, 2016.
[10] WEN A, XIE C, MAZHAR M, et al.Tetramethylpyrazine from adlay (Coix lacryma-jobi) biotransformation by Bacillus subtilis and its quality characteristics[J].Journal of Food Science and Technology, 2020, 57(11):4 092-4 102.
[11] 林榕姗.细菌型豆豉发酵机理及功能性研究[D].泰安:山东农业大学, 2012.
LIN R S.Studies on the fermentation mechanism and healthy functions of bacteria-fermented of bacteria-fermented douchi[D].Taian:Shandong Agricultural University, 2012.
[12] 郑战伟, 陈菁, 孙娟, 等.利用表面静态法发酵生产果醋的研究进展[J].农产品加工(学刊), 2011(3):24-26.
ZHENG Z W, CHEN J, SUN J, et al.Research progress of fermentation fruit vinegar by surface static method[J].Academic Periodical of Farm Products Processing, 2011(3):24-26.
[13] 吴晶晶.红曲霉(Monascus spp.)产生川芎嗪的发酵条件研究[D].杭州:浙江大学, 2011.
WU J J.Study on fermentation conditions of Monascus spp.producing tetramethypyrazine[D].Hangzhou:Zhejiang University, 2011.
[14] 郑环宇, 郎松彬, 崔月婷, 等.混菌株固态发酵高温豆粕工艺优化[J].中国食品学报, 2017, 17(9):116-124.
ZHENG H Y, LANG S B, CUI Y T, et al.Process optimization for solid-state fermentation of high temperture soybean meal by mixture strains[J].Journal of Chinese Institute of Food Science and Technology, 2017, 17(9):116-124.
[15] 彭真, 吴振强, 李运南, 等.红曲黄色素固态发酵条件的研究[J].现代食品科技, 2009, 25(6):669-673.
PENG Z, WU Z Q, LI Y N, et al.Study of solid-state fermentation by mohascus ahka for yellow pigment production[J].Modern Food Science and Technology, 2009, 25(6):669-673.
[16] 王辉.薏仁蒸煮特性及改进技术的研究[D].重庆:西南大学, 2014.
WANG H.Research on cooking characteristics and improved technology of coix seed[D].Chongqing:Southwest University, 2014.
[17] 郝飞, 吴群, 徐岩.枯草芽孢杆菌(Bacillus subtilis)发酵生产乙偶姻的pH调控策略[J].微生物学通报, 2013,40(6):921-927.
HAO F, WU Q, XU Y.Two-stage pH control strategy of acetoin production by Bacillus subtilis CCTCC M208157[J].Microbiology China, 2013,40(6):921-927.
[18] ZHU B, XU Y.Production of tetramethylpyrazine by batch culture of Bacillus subtilis with optimal pH control strategy[J].Journal of Industrial Microbiology & Biotechnology, 2010, 37(8):815-821.
[19] 赵菡.纳豆激酶热稳定性及pH稳定性的改造研究[D].无锡:江南大学, 2018.
ZHAO H.Study on enhanced thermal and pH stability of nottakinase[D].Wuxi:Jiangnan University, 2018.
[20] 付云, 赵谋明, 卢美杉, 等.枯草芽孢杆菌YA215发酵螺旋藻渣产抑菌活性的工艺[J].食品与发酵工业, 2020, 46(4):146-152.
FU Y, ZHAO M M, LU M S, et al.Fermentation optimization of antimicrobial spirulina residue from Bacillus subtilis YA215[J].Food and Fermentation Industries, 2020, 46(4):146-152.
[21] 朱兵峰.枯草杆菌两步法生产四甲基吡嗪的调控及机制的研究[D].无锡:江南大学, 2010.
ZHU B F.Fermentation control and mechanism of two-step tetramethylpyrazine production by Bacillus subtilis[D].Wuxi:Jiangnan University, 2010.
[22] CHEN J, CHEN Q, GUO Q, et al.Simultaneous determination of acetoin and tetramethylpyrazine in traditional vinegars by HPLC method[J].Food Chemistry, 2010, 122(4):1 247-1 252.
[23] NIU Y, YAO Z, XIAO Q, et al.Characterization of the key aroma compounds in different light aroma type Chinese liquors by GC-olfactometry, GC-FPD, quantitative measurements, and aroma recombination[J].Food Chemistry, 2017, 233:204-215.
[24] XIAO Z, ZHAO L, TIAN L, et al.GC-FID determination of tetramethylpyrazine and acetoin in vinegars and quantifying the dependence of tetramethylpyrazine on acetoin and ammonium[J].Food Chemistry, 2018, 239:726-732.
[25] 侯孝元, 顾如林, 梁文龙, 等.利用发酵法生产四甲基吡嗪研究进展[J].生物技术通报, 2016, 32(1):58-64.
HOU X Y, GU R L, LIANG W L, et al.Research progress on production of tetramethylpyrazine by fermentation[J].Biotechnology Bulletin, 2016, 32(1):58-64.
[26] XIAO Z, HOU X, LIU X, et al.Accelerated green process of tetramethylpyrazine production from glucose and diammonium phosphate[J].Biotechnology for Biofuels, 2014, 7(1):106.
