2022 01 v.33;No.105 123-130
产蛋白酶菌株的筛选及其在固态发酵豆粕中的应用
基金项目(Foundation):
浙江省重点研发计划项目(No.2021C04016);;
浙江省“领雁”研发攻关计划项目(No.2022C04020)
邮箱(Email):
mazheng1227@cjlu.edu.cn;
DOI:
中文作者单位:
中国计量大学生命科学学院浙江省生物计量及检验检疫技术重点实验室;浙江海洋大学水产学院;
摘要(Abstract):
目的:筛选高产蛋白酶菌株，探究微生物发酵对豆粕抗原蛋白的影响，以期提高豆粕在水产饲料中的使用价值。方法:从土壤样品中筛选出1株蛋白酶酶活为1 390.6±12.5 U/mL的菌株LX-6,经生理生化特性和16S rDNA分子鉴定为解淀粉芽孢杆菌(Bacillus amyloliquefaciens)。通过B.amyloliquefaciens LX-6不同时间发酵豆粕，分析SDS-PAGE中大分子抗原蛋白降解情况、发酵样品总蛋白以及水溶性蛋白含量。结果:在37℃、物料含50%水的条件下，发酵24 h豆粕中大分子抗原蛋白全部降解，48 h时水溶性蛋白含量达到最高44.45%。结论:B.amyloliquefaciens LX-6发酵后有效提高了豆粕的蛋白含量，此发酵豆粕有望今后开发成水产饲料。
关键词(KeyWords):
解淀粉芽孢杆菌;;蛋白酶;;固态发酵;;豆粕;;抗原蛋白
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参考文献
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[2] 周红蕾，李春玲，王贵平，等.大豆中抗营养因子及其去除方法概述[J].饲料工业，2006,3:23-26.ZHOU H,LI C L,WANG G P,et al.Overview of anti nutritional factors in soybean and their removal methods[J].Feed Industry,2006,3:23-26.
[3] 肇立春，李桂杰.论豆粕作饲料的质量鉴定[J].粮油加工与食品机械，2003,6:42-43.ZHAO L C,LI G J.On the quality identification of soybean meal as feed[J].Machinery For Cereals,Oil and Food Processing,2003,6:42-43.
[4] 过世东，胡建伟，赵娟，等.豆粕部分替代鱼粉、菜籽粕、棉籽粕对水产饲料的影响[J].粮食与饲料工业，2003(12):48-50.GUO S D,HU J W,ZHAO J,et al.Effects of soybean meal partially replacing fish meal,rapeseed meal and cottonseed meal on aquatic feed[J].Grain and Feed Industry,2003(12):48-50.
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[6] 张碧莹，杨蕊莲，张静，等.不同预处理方式对豆浆品质特性的影响[J].食品与发酵工业，2017,43(2):138-144.ZHANG B Y,YANG R L,ZHANG J,et al.Effects of different pretreatment methods on the quality characteristics of soybean milk[J].Food and Fermentation Industries,2017,43(2):138-144.
[7] 姚怡莎，谷旭，商方方，等.大豆和膨化大豆主要抗营养因子分析[J].中国农业科学，2016,49(11):2174-2182.YAO Y S,GU X,SHANG F F,et al.Analysis of main anti nutritional factors of soybean and expanded soybean[J].Scientia Agricultura Sinica,2016,49(11):2174-2182.
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[9] 魏凤仙，高方，李绍钰，等.膨化法与微生物发酵处理法对豆粕营养价值的影响[J].河南农业科学，2014,43(4):123-127.WEI F X,GAO F,LI S Y,et al.Effects of expansion and microbial fermentation on the nutritional value of soybean meal[J].Journal of Henan Agricultural Sciences,2014,43(4):123-127.
[10] 杨连玉，杨文艳.微生物发酵饲料的现状及展望[J].经济动物学报，2018,22(2):63-66.YANG L Y,YANG W Y.Present situation and prospect of microbial fermented feed[J].Journal of Economic Animals,2018,22(2):63-66.
[11] MUKHERJEE R,CHAKRABORTY R,DUTTA A.Role of fermentation in improving nutritional quality of soybean meal-a review[J].Asian-Australasian Journal of Animal Sciences,2016,29(11):1523-1529.
[12] SHRESTHA A,DAHAL N,NDUNGUTSE V.Bacillus fermentation of soybean:a review[J].Journal of Food Science and Technology Nepal,2013,6:1-9.
[13] YANG J,WU X,CHEN H,et al.A value-added approach to improve the nutritional quality of soybean meal byproduct:Enhancing its antioxidant activity through fermentation by Bacillus amyloliquefaciens SWJS22[J].Food Chemistry,2019,272:396-403.
[14] LIU Z,GUAN X,ZHONG X,et al.Bacillus velezensis DP-2 isolated from Douchi and its application in soybean meal fermentation[J].Journal of the Science of Food and Agriculture,2020,101(5):1-8.
[15] 吴宝昌，宋俊梅.枯草芽饱杆菌与米曲霉混合发酵制备豆粕饲料的研究[J].齐鲁工业大学学报：自然科学版，2010(2):18-21.WU B C,SONG J M.Preparation of soybean meal feed by mixed fermentation of Bacillus subtilis and Aspergillus oryzae[J].Journal of Qilu University of Technology:Natural Science Edition,2010(2):18-21.
[16] WONGPUTTISIN P,KHANONGNUCH C,KHONGBANTAD W,et al.Screening and selection of Bacillus spp.for fermented corticate soybean meal production[J].Journal of Applied Microbiology,2012,113(4):798-806.
[17] 郑丽，李达，牛红红，等.产蛋白酶芽孢杆菌的筛选及其发酵对豆粕的影响[J].食品科学，2019,40(20):185-191.ZHENG L,LI D,NIU H H,et al.Screening of protease producing Bacillus and its fermentation effect on soybean meal[J].Food Science,2019,40(20):185-191.
[18] 刘星，赵荣，李波，等.几种不同菌种发酵豆粕发酵过程指标对比[J].饲料博览，2020(1):20-23.LIU X,ZHAO R,LI B,et al.Comparison of fermentation process indexes of soybean meal fermented by several different strains[J].Feed Review,2020(1):20-23.
[19] 上海市酿造科学研究所.蛋白酶活力测定法：SB/T 10317-1999[S].北京：中国标准出版社，1999.
[20] 梁静南，刘一苇，谢家仪.制备细菌类单细胞生物扫描电镜样品方法的改进[J].电子显微学报，2013,32(3):276-278.LIANG J N,LIU Y W,XIE J Y.The improvement of bacteria sample preparation method for scanning electron microscope observation[J].Journal of Chinese Electron Microscopy Society,2013,32(3):276-278.
[21] 东秀珠，蔡妙英.常见细菌系统鉴定手册[M].北京：科学出版社，2001:364-399.
[22] 广东省微生物分析检测中心.饲用微生物制剂中枯草芽孢杆菌的检测：GB/T 26428-2010[S].北京：中国标准出版社，2011.
[23] 张文刚，张妤，孙冰清，等.凝胶电泳法(SDS-PAGE)测定饲料添加剂溶菌酶二聚体的含量[J].养殖与饲料，2021,20(7):6-8.ZHANG W G,ZHANG Y,SUN B Q,et al.Gel electrophoresis (SDS-PAGE) was used to determine the content of lysozyme two dimer in feed additive[J].Animals Breeding and Feed,2021,20(7):6-8.
[24] 张琳静.凯氏定氮法测定豆粕中粗蛋白含量[J].兽医导刊，2017(6):213-214.ZHANG L J.Determination of crude protein content in soybean meal by Kjeldahl method[J].Veterinary Orientation,2017(6):213-214.
[25] 国家粮食局.大豆储存品质判定规则：GB/T 31785-2015[S].北京：中国标准出版社，2015.
[26] 陈菲.一株蛋白酶产生菌的鉴定、酶性质研究和表达载体构建[D].郑州：河南工业大学，2018:19-40.CHEN F.IdentificationEnzyme Properties and Expression Vector Construction of a Protease Producing Strain[D].Zhengzhou:Henan University of Technology,2018:19-40.
[27] LEONEL O,OLMOSSOTO J.The functional property of Bacillus for shrimp feeds-ScienceDirect[J].Food Microbiology,2006,23(6):519-525.
[28] 陈瑜，孔海深.伯杰鉴定细菌学手册第9版简介[J].国外医学(微生物学分册),1995,18(6):32-48.CHEN Y,KONG H S.Berger’s Handbook of identification bacteriology,9th Edition[J].Foreign Medicine(Microbiology),1995,18(6):32-48.
[29] SUMPAVAPOL P,TONGYONK L.Bacillus siamensis sp.nov.isolated from salted crab (poo-khem) in Thailand[J].International Journal of Systematic and Evolutionary Microbiology,2010,60(10):2364-2370.
[30] CHI C,CHO S.Improvement of bioactivity of soybean meal by solid-state fermentation with Bacillus amyloliquefaciens versus Lactobacillus spp.and Saccharomyces cerevisiae[J].LWT-Food Science and Technology,2016,68:619-625.
[31] 胡毅恒，何华锋，黄光荣，等.嗜热芽孢杆菌产耐热蛋白酶稳定性的研究[J].中国计量学院学报，2009,20(2):149-153.HU Y H,HE H F,HUANG G R,et al.Stability of a thermostable protease from a thermophilic Bacillus[J].Journal of China University of Metrology,2009,20(2):149-153.
[32] 吝常华，刘国华，常文环，等.豆粕微生物固态发酵工艺优化及其营养物质含量变化[J].动物营养学报，2018,30(7):2749-2762.SHI C H,LIU G H,CHANG W H,et al.Optimization of soybean meal microbial solid-state fermentation process and changes of its nutrient content[J].Chinese Journal of Animal Nutrition,2018,30(7):2749-2762.
[33] PARk J,LEE N,KIM B,et al.Characterization of traditionally fermented Korean soybean paste,eoyukjang,and isolation of its microorganisms[J].Food Science & Biotechnology,2010,19(2):425-430.
[34] 刘慧珍.豆粕适度酶解研究[D].无锡：江南大学，2007:38-44.LIU H Z.Study on Moderate Enzymatic Hydrolysis of Soybean Meal[D].Wuxi:Jiangnan University,2007:38-44.
基本信息:
DOI：
中图分类号:S963
引用信息:
[1]李慧杰,林恒伊,黄心怡等.产蛋白酶菌株的筛选及其在固态发酵豆粕中的应用[J].中国计量大学学报,2022,33(01):123-130.
基金信息:
浙江省重点研发计划项目(No.2021C04016);; 浙江省“领雁”研发攻关计划项目(No.2022C04020)
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