Food and Fermentation Industries >

2025 , Vol. 51 >Issue 20: 112 - 121

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.042012

Characterization of enzymatic properties of proteolytic enzymes produced by Bacillus amyloliquefaciens BS5582 and application in soy sauce fermentation

  • YANG Shijing ,
  • CHEN Huating ,
  • LIU Chunfeng ,
  • NIU Chengtuo ,
  • LI Qi
Expand
  • 1(Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China)
    2(School of Biotechnology, Jiangnan University, Wuxi 214122, China)

Received date: 2024-12-31

  Revised date: 2025-03-07

  Online published: 2025-10-27

Fold

Abstract

Proteolytic enzymes play crucial roles in soy sauce fermentation.To improve the utilization rate of raw materials and soy sauce quality, this research aimed to characterize the enzymatic properties of proteolytic enzymes produced by Bacillus amyloliquefaciens BS5582 and evaluate its application in soy sauce fermentation.The optimal temperature and optimal pH value of enzymes were 50 ℃ and pH 7.5, respectively.The half-life value at pH 6.0 reached 86.93 min, which was better than two commercial enzymes.Moreover, this enzyme also had a relatively high tolerance to high salinity since it could still maintain around 50% activity after treatment at 140 g/L salinity for 15 days.Finally, the proteolytic enzymes produced by the BS5582 strain were applied in the initial stage of soy sauce fermentation.Results showed that the addition of BS5582 enzymes could significantly increase the concentrations of amino acid nitrogen and the quantity and concentration of volatile flavor substances in soy sauce.The soy sauce samples fermented with the addition of proteolytic enzymes also had a stronger fruity aroma, sauce aroma and smoky aroma, which endowed soy sauce with a more pleasant aroma and more mellow taste.The results obtained in this study showed that the proteolytic enzymes produced by B.amyloliquefaciens BS5582 had the good enzymatic ability and benefited the flavor and quality of soy sauce, indicating the enzyme had potential application value in soy sauce production.

Key words： Bacillus amyloliquefaciens BS5582; proteolytic enzymes; enzymatic properties; soy sauce

Cite this article

YANG Shijing , CHEN Huating , LIU Chunfeng , NIU Chengtuo , LI Qi . Characterization of enzymatic properties of proteolytic enzymes produced by Bacillus amyloliquefaciens BS5582 and application in soy sauce fermentation[J]. Food and Fermentation Industries, 2025 , 51(20) : 112 -121 . DOI: 10.13995/j.cnki.11-1802/ts.042012

References

[1] SASSI S, AL QADR IMAD, WAN-MOHTAR W A, JAMALUDIN N S, et al.Recent progress and advances in soy sauce production technologies:A review[J].Journal of Food Processing and Preservation, 2021, 45(10):e15799.
[2] SONG P, ZHANG X, WANG S H, et al.Microbial proteases and their applications[J].Frontiers in Microbiology, 2023, 14:1236368.
[3] 赵雪, 张展开, 张智宏, 等.酱油酿造过程中微生物及生物酶的研究进展[J].现代食品科技, 2024, 40(2):329-337.
ZHAO X, ZHANG Z K ZHANG Z H, et al.Progress on microorganisms and biological enzymes during soy sauce fermentation[J].Modern Food Science and Technology, 2024, 40(2):329-337.
[4] PENG M Y, LIU J Y, HUANG Y, et al.Effects of a mixed koji culture of Aspergillus oryzae HG-26 and Aspergillus niger HG-35 on the levels of enzymes, antioxidants and phenolic compounds in soy sauce during the fermentation process[J].International Journal of Food Science & Technology, 2017, 52(7):1585-1593.
[5] LIU Y H, SUN G R, LI J Y, et al.Starter molds and multi-enzyme catalysis in koji fermentation of soy sauce brewing:A review[J].Food Research International, 2024, 184:114273.
[6] 冯纬, 周利南, 刘晔, 等.酿造酱油发酵过程及其相关酶系[J].价值工程, 2018, 37(12):165-166.
FENG W, ZHOU L N, LIU Y, et al.The fermentation period of brewing soy sauce and the enzyme system during soy sauce fermentation[J].Value Engineering, 2018, 37(12):165-166.
[7] CHEN C F, HOU S, WU C Z, et al.Improving protein utilization and fermentation quality of soy sauce by adding protease[J].Journal of Food Composition and Analysis, 2023, 121:105399.
[8] 任涛, 钮成拓, 郑飞云, 等.淀粉液化芽孢杆菌BS5582产蛋白酶的发酵条件优化、酶学性质分析及其在牛皮脱毛中的应用[J].食品与发酵工业, 2024, 50(22):106-114.
REN T, NIU C T, ZHENG F Y, et al.Fermentation optimization and enzymatic property analysis of proteases produced by Bacillus amyloliquefaciens BS5582 and its application in cattle skin dehairing[J].Food and Fermentation Industries, 2024, 50(22):106-114.
[9] 黄佳玲, 刘建华, 刘艳梅, 等.种曲接种量对酱油制曲及发酵的影响[J].食品安全导刊, 2023(25):143-147;151.
HUANG J L, LIU J H, LIU Y M, et al.Effect of inoculum size of seed koji on soy sauce koji-making and the fermentation[J].China Food Safety Magazine, 2023(25):143-147;151.
[10] 康辉. 固态发酵及酶解对豆粕中鲜味肽的释放影响研究[D].广州:华南理工大学, 2023.
KANG H.Effects of solid-state fermentation and enzymatic hydrolysis on releasing umami peptides from soybean meal[D].Guangzhou:South China University of Technology, 2023.
[11] 赵杨杨. 高盐稀态发酵法制备香菇酱油的研究[D].烟台:烟台大学, 2023.
ZHAO Y Y.Study on preparation technology of Lentinus edodes soy sauce by high-salt dilute fermentation[D].Yantai:Yantai University, 2023.
[12] 邢先雷. 生料发酵豆瓣酱微生物群落形成规律研究[D].无锡:江南大学, 2024.
XING X L.Analysis of microbial community dynamics during raw materials based broad bean paste fermentation[D].Wuxi:Jiangnan University, 2024.
[13] 苗春雷, 朱娅媛, 黄卓权, 等.酱油电渗析液用于酱油制曲的研究[J].中国酿造, 2024, 43(11):180-186.
MIAO C L, ZHU Y Y, HUANG Z Q, et al.Application of soy sauce electrodialysis brine in the koji-making of soy sauce[J].China Brewing, 2024, 43(11):180-186.
[14] 谭戈, 肖亮琴, 吴昌正, 等.基于GC-MS分析不同酿造周期酱油产品的挥发性风味物质[J].中国调味品, 2023, 48(4):161-165.
TAN G, XIAO L Q, WU C Z, et al.Analysis of volatile flavor compounds in soy sauce products with different brewing periods based on GC-MS[J].China Condiment, 2023, 48(4):161-165.
[15] LIOE H N, WADA K, AOKI T, et al.Chemical and sensory characteristics of low molecular weight fractions obtained from three types of Japanese soy sauce (shoyu)-Koikuchi, tamari and shiro shoyu[J].Food Chemistry, 2007, 100(4):1669-1677.
[16] 黄毅, 锁然, 李娜.酱油中氨基酸的组成与分析[J].中国调味品, 2012, 37(3):101-104;107.
HUANG Y, SUO R, LI N.Composition and analysis of amino acids in soy sauce[J].China Condiment, 2012, 37(3):101-104;107.
[17] ZHANG L Q, ZHOU R Q, CUI R Y, et al.Characterizing soy sauce moromi manufactured by high-salt dilute-state and low-salt solid-state fermentation using multiphase analyzing methods[J].Journal of Food Science, 2016, 81(11):C2639-C2646.
[18] LIOE H N, SELAMAT J, YASUDA M.Soy sauce and its umami taste:A link from the past to current situation[J].Journal of Food Science, 2010, 75(3):R71-R76.
[19] GAO L H, LIU T, AN X J, et al.Analysis of volatile flavor compounds influencing Chinese-type soy sauces using GC-MS combined with HS-SPME and discrimination with electronic nose[J].Journal of Food Science and Technology, 2017, 54(1):130-143.
[20] DEVANTHI P V P, GKATZIONIS K.Soy sauce fermentation:Microorganisms, aroma formation, and process modification[J].Food Research International, 2019, 120:364-374.
[21] DIEZ-SIMON C, EICHELSHEIM C, MUMM R, et al.Chemical and sensory characteristics of soy sauce:A review[J].Journal of Agricultural and Food Chemistry, 2020, 68(42):11612-11630.
[22] KANEKO S, KUMAZAWA K, NISHIMURA O.Studies on the key aroma compounds in raw (unheated) and heated Japanese soy sauce[J].Journal of Agricultural and Food Chemistry, 2013, 61(14):3396-3402.
[23] WANG P P, KAN Q X, YANG L X, et al.Characterization of the key aroma compounds in soy sauce by gas chromatography-mass spectrometry-olfactometry, headspace-gas chromatography-ion mobility spectrometry, odor activity value, and aroma recombination and omission analysis[J].Food Chemistry, 2023, 419:135995.
[24] WU W Y, CHEN T, ZHAO M M, et al.Effect of co-inoculation of different halophilic bacteria and yeast on the flavor of fermented soy sauce[J].Food Bioscience, 2023, 51:102292.
Options
Outlines

/

〈 〉

Powered by Beijing Magtech Co. Ltd,.