芽孢杆菌高产活性物质与诱变育种
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摘要
芽孢杆菌属细菌在农业、工业、医药、环境等各个领域发挥重要作用。本综述介绍了芽孢杆菌的主要活性物质和选育方法。分别从酶类物质、抗菌脂肽类物质、其他类型物质3个方面对芽孢杆菌主要活性产物的基本特征和功能做了详细介绍。概述了自然选育、诱变选育、原生质体选育、基因组改组选育和杂交选育5种常见的选育方法和基本原理。提供了15种芽孢杆菌菌株获得高产活性物质的诱变方法,其中1株芽孢杆菌经2次紫外诱变和1次DES处理后,产中性蛋白酶的能力提高到7 307 U/m L。这些诱变方法为今后快速获得相关活性物质提供了理论指导。
Bacillus bacteria play an important role in agriculture, industry, medicine, environment and other fields. This paper described the main active products and breeding methods of Bacillus. The basic characteristics and functions of the main active products of Bacillus were described in detail from three aspects: enzyme substances, antimicrobial lipopeptides and other types of substances. 5 main breeding methods and basic principles were introduced of natural selection, mutation breeding, protoplast selection, genome shuffling and hybrid breeding. At the same time, this paper provided the mutation methods of 15 Bacillus strains to obtain high yield active substances, The ability of producing a neutral protease was increased to 7 307 U/m L after 2 times of UV mutagenesis and once DES treatment. These mutagenesis methods provided theoretical guidance for the rapid acquisition of related active substances in the future.
Bacillus bacteria play an important role in agriculture, industry, medicine, environment and other fields. This paper described the main active products and breeding methods of Bacillus. The basic characteristics and functions of the main active products of Bacillus were described in detail from three aspects: enzyme substances, antimicrobial lipopeptides and other types of substances. 5 main breeding methods and basic principles were introduced of natural selection, mutation breeding, protoplast selection, genome shuffling and hybrid breeding. At the same time, this paper provided the mutation methods of 15 Bacillus strains to obtain high yield active substances, The ability of producing a neutral protease was increased to 7 307 U/m L after 2 times of UV mutagenesis and once DES treatment. These mutagenesis methods provided theoretical guidance for the rapid acquisition of related active substances in the future.
引文
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Bai D.,Long L.,and Zhu N.S.,2016,Mutation breeding of a isolated Bacillus subtilis var.niger for producing the1-Deoxynojirimycin,Fudan Xuebao(Journal of Fudan University),55(1):104-111(白豆,龙玲,朱乃硕,2016,一株产1-脱氧野尻霉素(DNJ)枯草芽孢杆菌黑色变种的诱变育种研究,复旦学报,55(1):104-111)
Balaji L.,and Jayaraman G.,2014,Metalion activated lipase from halotolerant Bacillus sp.VITL8 displays broader operational range,Inter.J.Biol.Macromol.,67:380-386
Bao Z.G.,2013,Study on breeding of phytase-producing Bacillus subtilis and its fermentation products from cottonseed meal source,Thesis for M.S.,Shihezi University,Supervisor:Zhang W.J.,pp.17-23(鲍振国,2013,产植酸酶枯草芽孢杆菌的选育及其棉粕源发酵产物的研究,硕士学位论文,石河子大学,导师:张文举,pp.17-23)
Bussamara R.,Fuentefria A.M.,and de Oliveira E.S.,2010,Isolation of alipase-secreting yeast for enzyme production in apilot-plant scale batch fermentation,Bioresource Technology,101(1):268-275
Dong H.P.,Wang Y.J.,and Zheng Y.G.,2010,Enantioselective hydrolysis of diethyl 3-hydroxyglutarate to ethyl(s)-3-hy droxyglutarate by immobilized Candida antarctica lipase,Journal of Molecular Catalysis B:Enzymatic,66(1-2):90-94
Gao X.W.,Yao S.Y.,and Wang J.S.,2004,Purification and identification of surfactin isoforms produced by Bacillus subtilis B2 Strain,Weishengwu Xuebao(Acta Microbiologica Sinica),43(5):647-652(高学文,姚仕义,王金生,2004,枯草芽孢杆菌B2菌株产生的表面活性素变异体的纯化和鉴定,微生物学报,43(5):647-652)
Guan M.L.,Li X.P.,Yang M.,Zhang Z.M.,Zhang W.B.,and Shi X.X.,2016,Study on mutation breeding of Bacillus amyloliquefaciens producing milk-clotting enzyme,Shiping Gongye Keji(Science and Technology of Food Industry),37(3):156-159(关明玲,李学朋,杨敏,张忠明,张卫兵,师希雄,2016,产凝乳酶解淀粉芽孢杆菌的诱变选育,食品工业科技,37(3):156-159)
Gui C.Y.,2010,Study on strain screening of haloduruc and alkaline Bacillus sp.and its properties of cellulase,Thesis for M.S.,Shandong Agricultural University,Supervisor:Chen Y.L.,pp.5-10(桂春燕,2010,耐盐碱产纤维素酶芽孢杆菌的选育及酶学特性的研究,硕士学位论文,山东农业大学,导师:陈义伦,pp.5-10)
Han K.F.,2013,Mutation breeding and purification ofβ-mannanase from an endophyic Paenibacillus sp CH-3,Thesis for M.S.,Henan Normal University,Supervisor:Zhang J.X.,pp.9-21(韩科芳,2013,β-甘露聚糖酶内生类芽孢杆菌诱变选育及酶的纯化研究,硕士学位论文,河南师范大学,导师:张建新,pp.9-21)
Hu X.R.,2006,Studies on the application of interspecific protoplast fusion techniques to the breeding of Bacillus subtilis with high cellulose activity,Thesis for M.S.,Northwest University,Supervisor:Chen W.L.,pp.3-8(胡欣荣,2006,利用原生质体融合技术选育高纤维素酶活枯草芽孢杆菌的研究,硕士学位论文,西北大学,导师:陈五岭,pp.3-8)
Jing X.Q.,2013,Study on the breeding of Bacillus natto with high yieldγ-PGA and fermentation characteristics,Thesis for M.S.,Anhui Polytechnic University,Supervisor:Zhang Q.Q.,pp.2-5(金鑫强,2013,高产聚谷氨酸纳豆芽孢杆菌选育及其发酵特性研究,硕士学位论文,安徽工程大学,导师:张庆庆,pp.2-5)
Ju H.W.,Jiang L.Z.,Wang Q.J.,and Zou Y.,2007,Study on the breeding of Bacillus subtilis strain with high alkaline protease and its hydrolysis conditions,Shiping Gongye Keji(Science and Technology of Food Industry),28(8):109-111(鞠华伟,江连洲,王秋京,邹阳,2007,高产碱性蛋白酶枯草芽孢杆菌菌株选育及其水解条件的研究,食品工业科技,28(8):109-111)
Li X.Y.,2012,Screening high yield Bacillus cereus of amylaseproducing strain and studying the characterization of the amylase,Thesis for M.S.,Jilin Agricultural University,Supervisor:Chen G.,pp.1-28(李旭媛,2012,产淀粉酶蜡状芽孢杆菌的诱变选育及酶学性质的研究,硕士学位论文,吉林农业大学,导师:陈光,pp.1-28)
Liu D.,2010,Characterizations of chitinases from Bacillus thuringiensis subsp colmeri 15A3 and identification of promoter sequence of chi B,Dissertation for Ph.D.,Nankai University,Supervisor:Chen Y.H.,pp.2-6(刘东,2010,苏云金芽孢杆菌几丁质酶特性分析及其基因启动子序列的确定,博士学位论文,南开大学,导师:陈月华,pp.2-6)
Liu H.L.,2014,Mutation breeding and laboratory evolution of thermotoletant Bacillus coagulans for L-Lactic acid production,Thesis for M.S.,East China University of Science and Technology,Supervisor:Zhang S.L.,pp.1-6(刘慧兰,2014,产乳酸耐高温凝结芽孢杆菌的诱变及进化选育,硕士学位论文,华东理工大学,导师:张嗣良,pp.1-6)
Michelson T.,Kask K.,Jogi E.,and Talpsep E.,2006,(+)-Lactic acid producer Bacillus coagulans SIM-7 DSM 14043 and its comparison with Lactobacillus delbrueckii sp.lactis DSM20073,Enzyme and Microbial Technology,39(4):861-867
Shi X.K.,Wang X.R.,and Liu D.P.,2005,Genome shuffling accelerate the process of microbial breeding,Nongye Yu Jishu(Agriculture and Technology),25(1):145-147(史晓昆,王秀然,刘东波,2005,Genome Shuffling技术在微生物遗传育种中的应用,农业与技术,25(1):145-147)
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Zhang L.,2013,Breeding of Bacillus subtilis BS1 for improving surfactin yield,Thesis for M.S.,Nanjing Agricultural University,Supervisor:Lu Z.X.,pp.1-8(张莉,2013,枯草芽孢杆菌BS1 Surfactin高产菌株的选育研究,硕士学位论文,南京农业大学,导师:陆兆新,pp.1-8)
Zhang M.,Hu X.,Wan J.Y.,Lei X.Y.,and Zhao D.P.,2010,Mutagenesis breeding of Bacillus subtilis 515 strain for chitinase production and study on its fermentation condition,Zhongguo Nongxue Tongbao(Chinese Agricultural Science Bulletin),26(11):279-283(张敏,胡晓,万津瑜,雷欣雨,赵丹萍,2010,高产几丁质酶的枯草芽孢杆菌诱变育种及发酵条件研究,中国农学通报,26(11):279-283)
Zhang S.L.,2006,The screening and incubation of Bacillus sp.stain producing alginate lyase and researches on its properties,Thesis for M.S.,Ocean University of China,Supervisor:Guan B.,pp.16-44(张书利,2006,产褐藻胶裂解酶芽孢杆菌的选育及其产酶性质的研究,硕士学位论文,中国海洋大学,导师:管斌,pp.16-44)
Zhao Y.T.,2009,Mutagenesis breeding and fermentation condi tions optimization of Bacillus subtilis BL-1,Thesis for M.S.,Heilongjiang University,Supervisor:Du C.M.,pp.10-15(赵云彤,2009,侧芽孢杆菌BL-1的诱变选育及发酵条件优化,硕士学位论文,黑龙江大学,导师:杜春梅,pp.10-15)
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