青藏高原土壤低温产蛋白酶菌株的筛选及酶学性质研究
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摘要
从青藏高原土壤样品分离保藏的菌种中筛选到低温条件下产蛋白酶菌株LS20-2,对其进行生理生化试验以及分子生物学鉴定,并对其发酵液进行酶学性质初步研究。结果表明,该菌被鉴定为皮氏类芽孢杆菌(Paenibacillus peoriae),产蛋白酶的最适反应p H值为7.0,最适温度为40℃。在p H5.0~7.0之间、温度0~40℃时酶活稳定,金属离子Fe~(2+)、Zn~(2+)及Triten X-100能抑制蛋白酶活性,金属离子Mn~(2+)及表面活性剂Tween-20、Tween-80能提高蛋白酶活性。
A low temperature protease-producing strain,named as LS20-2,was screened from the Qinghai-Tibet Plateau soil samples.After physiological and biochemical identification and molecular biology,and the enzymatic properties of the fermentation broth was preliminarily studied.Results showed that the strain was identified as Paenibacillus peoriae,the optimal p H and temperature for the protease were 7.0 and 40℃,respectively.The protease activity was stable at p H 5.0-7.0 and temperature 0-40℃.Metal ions Fe~(2+),Zn~(2+)and Triten X-100 could inhibit protease activity,Mn~(2+)and surfactant Tween-20,Tween-80 could improve the activity of protease.
A low temperature protease-producing strain,named as LS20-2,was screened from the Qinghai-Tibet Plateau soil samples.After physiological and biochemical identification and molecular biology,and the enzymatic properties of the fermentation broth was preliminarily studied.Results showed that the strain was identified as Paenibacillus peoriae,the optimal p H and temperature for the protease were 7.0 and 40℃,respectively.The protease activity was stable at p H 5.0-7.0 and temperature 0-40℃.Metal ions Fe~(2+),Zn~(2+)and Triten X-100 could inhibit protease activity,Mn~(2+)and surfactant Tween-20,Tween-80 could improve the activity of protease.
引文
[1]刘海军,乐超银,邵伟,等.一株高产蛋白酶芽孢杆菌的鉴定[J].中国酿造,2009,28(9):18-20.
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[10]陈秀兰,张玉忠,高培基,等.渤海湾浅表海水中产低温蛋白酶适冷菌的筛选[J].海洋科学,2000,24(9):42-45.
[11]倪永清,顾燕玲,史学伟,等.天山一号冰川底部沉积层产蛋白酶耐低温菌株的筛选及其系统发育[J].微生物学报,2013,53(2):164-172.
[12]SINGH S K,SINGH S K,TRIPATHI V R,et al.A novel psychrotrophic,solvent tolerant Pseudomonas putida SKG-1 and solvent stability of its psychrothermoalkalistable protease[J].Process Biochem,2014,46(7):1430-1435.
[13]陈明霞,李和阳,陈维维,等.68株北极产蛋白酶菌株的筛选、鉴定以及部分酶学性质[J].微生物学报,2013,53(7):702-709.
[14]XIONG H,SONG L,XU Y,et al.Characterization of proteolytic bacteria from the Aleutian deep-sea and their protease[J].J Ind Microbiol Biot,2007,34(1):63-71.
[15]权淑静,马焕,刘德海,等.青藏高原土壤产α-淀粉酶菌株的分离、鉴定及产酶特性研究[J].河南科学,2015,33(3):384-388.
[16]马焕,权淑静,刘德海,等.产木聚糖酶菌株的筛选、鉴定及其酶学性质研究[J].中国酿造,2016,35(12):123-128.
[17]东秀珠,蔡妙英.常见细菌系统鉴定手册[M].北京:科学出版社,2001:364-390.
[2]RAO M B,TANKSALE A M,GHATGE M S,et al.Molecular and biotechnological aspects of microbial proteases[J].Microbiol Mol Biol Rev,1998,62(3):597-635.
[3]YANG J K,SHIH I L,TZENG Y M,et al.Production and purification of protease from Bacillus subtilis that can deproteinize crustacean wastes[J].Enzyme Microb Technol,2000,26(5-6):406-413.
[4]VARELA H,FERRARI M D,BELOBRAJDIC L,et al.Skin unhairing proteases of Bacillus subtilis:production and partial characterization[J].Biotechnol Lett,1997,19(8):755-758.
[5]高兆建,陈尚龙,巫有华,等.产低温弹性蛋白酶菌株的筛选及酶学特性分析[J].食品与生物技术学报,2012,3(10):1106-1111.
[6]JOSHI S,SATYANARAYANA T.Biotechnology of cold-active proteases[J].Biology,2013,2(2):755-783.
[7]莫清珊,张会图,田耀,等.低温蛋白酶产生菌的筛选及其酶学性质的初步研究[J].天津科技大学学报,2015,30(3):19-23.
[8]KASANA R C.Proteases from psychrotrophs:An overview[J].Crit Rev Microbiol,2010,36(2):134-145.
[9]MAEGESIN R,DIEPLINGER H,HOFMANN J,et al.A cold-active extracellular metalloprotease from Pedobacter cryoconitis:production and properties[J].Res Microbiol,2005,156(4):499-505.
[10]陈秀兰,张玉忠,高培基,等.渤海湾浅表海水中产低温蛋白酶适冷菌的筛选[J].海洋科学,2000,24(9):42-45.
[11]倪永清,顾燕玲,史学伟,等.天山一号冰川底部沉积层产蛋白酶耐低温菌株的筛选及其系统发育[J].微生物学报,2013,53(2):164-172.
[12]SINGH S K,SINGH S K,TRIPATHI V R,et al.A novel psychrotrophic,solvent tolerant Pseudomonas putida SKG-1 and solvent stability of its psychrothermoalkalistable protease[J].Process Biochem,2014,46(7):1430-1435.
[13]陈明霞,李和阳,陈维维,等.68株北极产蛋白酶菌株的筛选、鉴定以及部分酶学性质[J].微生物学报,2013,53(7):702-709.
[14]XIONG H,SONG L,XU Y,et al.Characterization of proteolytic bacteria from the Aleutian deep-sea and their protease[J].J Ind Microbiol Biot,2007,34(1):63-71.
[15]权淑静,马焕,刘德海,等.青藏高原土壤产α-淀粉酶菌株的分离、鉴定及产酶特性研究[J].河南科学,2015,33(3):384-388.
[16]马焕,权淑静,刘德海,等.产木聚糖酶菌株的筛选、鉴定及其酶学性质研究[J].中国酿造,2016,35(12):123-128.
[17]东秀珠,蔡妙英.常见细菌系统鉴定手册[M].北京:科学出版社,2001:364-390.