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地热田碱性蛋白酶菌株的选育与酶性质的研究
引用本文:黄家福,潘裕添,陈俊玉,林荣榕,徐宝丹,余金富. 地热田碱性蛋白酶菌株的选育与酶性质的研究[J]. 漳州师院学报, 2011, 0(1): 48-55
作者姓名:黄家福  潘裕添  陈俊玉  林荣榕  徐宝丹  余金富
作者单位:[1]福建漳州师范学院生物科学与技术系,福建漳州363000 [2]东山出入境检验检疫局,福建漳州363401 [3]福建福州市第四十中学,福建福州350001 [4]福建漳州科人生物技术开发有限公司,福建漳州363000
基金项目:福建省自然科学基金计划项目(2010J05088)
摘    要:从温泉附近土样中成功分离到一株高温碱性蛋白酶产生菌株H-1,经初步鉴定为芽孢杆菌(Bacillus.)菌株H-1最佳产酶的发酵时间为67.5h,该酶在30~70℃酶活性随温度升高而增强,70℃酶活性达到最高,在pH7.0~11.0范围内较稳定,从菌株H-1液体发酵液中提取粗酶液,经CMC-I阳离子交换层析和SepHadex G-75凝胶层析,得到单一组分;通过Native-PAGE、SDS-PAGE电泳鉴定,粗酶至少含有3种碱性蛋白酶同工酶;单组分碱性蛋白酶的比活力为3875U,相对分子量约为31KDa.

关 键 词:碱性蛋白酶  同工酶  阳离子交换层析  凝胶层析  地热田

Breeding of Alkaline Protease Producing Strain from Geothermal Field and Properties of the Enzyme
HUANG Jia-fu,PAN Yu-tian,CHEN Jun-yu,LIN Rong-rong,XU Bao-dan,YU Jin-fu. Breeding of Alkaline Protease Producing Strain from Geothermal Field and Properties of the Enzyme[J]. Journal of ZhangZhou Teachers College(Philosophy & Social Sciences), 2011, 0(1): 48-55
Authors:HUANG Jia-fu  PAN Yu-tian  CHEN Jun-yu  LIN Rong-rong  XU Bao-dan  YU Jin-fu
Affiliation:1.Department of Biology Science and Technology,Zhanzhou Normal University,Zhangzhou,Fujian 363000,China; 2.Dongshan Entry-Exit Inspection and Quarantine Bureau of P.R.C,Zhangzhou,Fujian 363401,China;3.The Fortieth School of Fuzhou,Fuzhou,Fujian 350001,China;4.Zhangzhou Keren Biotechnology Deveplopment Co.,Ltd.,,Zhangzhou,Fujian 363000,China)
Abstract:A thermotolerant alkaline protease producing strain,H?-1,was isolated from soil sample in the neighborhood of hot spring in Zhangzhou,Fujian province.It is primarily examined as Bacillus.The best time of producting protease was 67.5 hours after the strain H-1 was cultured.The activity of this thermotolerant alkaline protease was rising gradually from 30℃ to 70℃,reaching the climax at about 70℃,keeping stable under pH 7.0~11.0.The single component was purified from the crude culture filtrate of the strain H-1 by cation exchange chromatograpHy on CMC-I and gel filtration chromatograpHy on SepHadex G-75.The result of Native-PAGE and SDS-PAGE showed that at least three kinds of isoenzymes existed in the crude protease,the rate activity of this single component was 3875U,and its molecular weight was about 31KDa.
Keywords:Alkaline Protease  Isoenzymes  cation exchange chromatograpHy  congeal glue layer  geothermal field
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