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中国精品科技期刊2020
刘唤明, 张芷欣, 洪鹏志, 周春霞, 杨萍, 陈康健, 吴金红. 耐高温蛋白酶产生菌的筛选及酶学特性的初步研究[J]. 食品工业科技, 2017, (03): 133-136. DOI: 10.13386/j.issn1002-0306.2017.03.017
引用本文: 刘唤明, 张芷欣, 洪鹏志, 周春霞, 杨萍, 陈康健, 吴金红. 耐高温蛋白酶产生菌的筛选及酶学特性的初步研究[J]. 食品工业科技, 2017, (03): 133-136. DOI: 10.13386/j.issn1002-0306.2017.03.017
LIU Huan-ming, ZHANG Zhi-xin, HONG Peng-zhi, ZHOU Chun-xia, YANG Ping, CHEN Kang-jian, WU Jin-hong. Isolation of microbial strain producing thermostable protease and characterization of the enzyme[J]. Science and Technology of Food Industry, 2017, (03): 133-136. DOI: 10.13386/j.issn1002-0306.2017.03.017
Citation: LIU Huan-ming, ZHANG Zhi-xin, HONG Peng-zhi, ZHOU Chun-xia, YANG Ping, CHEN Kang-jian, WU Jin-hong. Isolation of microbial strain producing thermostable protease and characterization of the enzyme[J]. Science and Technology of Food Industry, 2017, (03): 133-136. DOI: 10.13386/j.issn1002-0306.2017.03.017

耐高温蛋白酶产生菌的筛选及酶学特性的初步研究

Isolation of microbial strain producing thermostable protease and characterization of the enzyme

  • 摘要: 为了提供可用于水产饲料添加剂用途的蛋白酶,从玛珥湖环境中筛选出耐高温蛋白酶产生菌株。通过牛奶平板筛选的方法从土壤中分离获得30株产蛋白酶菌株,并通过摇瓶发酵复筛的方法筛选产酶最高的菌株HL-3,其发酵液的蛋白酶活性高达399.2 U/m L。通过生理生化特征鉴定、16S r DNA基因序列和系统发育分析,初步鉴定菌株HL-3为地衣芽孢杆菌。菌株HL-3所产蛋白酶的最适作用温度为70℃,最适p H为7.0,在90℃保温10 min后酶活力仍可保留75.12%。在离子浓度为1.0×10-3mol/L时,Cu2+和Mn2+对该酶有较强的抑制作用,Fe2+、Zn2+、Ca2+对该酶有激活作用,Mg2+和K+对该酶活性几乎没有影响。预期该菌株产的蛋白酶具有用于水产饲料添加剂的潜力。 

     

    Abstract: In order to provide protease for potential use in aquaculture feed industry,the strain producing thermostable protease was screened from the environment of maar lake.Thirty strains were isolated from soil by milk plates.Strain HL-3,which showed the highest production,was screened according to fermenting in shaking flasks,and the enzymatic activity of protease in culture solution of HL-3 was 399.2 U / mL.Based on physiological and biochemical identification,16S rDNA sequence and phylogenetic analysis,the strain HL- 3 was preliminarily identified as Bacillus licheniformis. The protease activity preformed optimally at70 ℃ and p H7.0. The protease retained 75.12% of its original activity after incubation for 10 min at 90 ℃. When ion concentration reached 1.0 × 10~(-3)mol / L,metal ions of Cu~(2+) and Mn~2+) would strongly inhibit the protease activity,metal ions of Fe~(2+),Zn~(2+)and Ca~(2+) would activate the protease activity,metal ions of Mg~(2+) and K~+would not affect the protease activity.The protease produced by strain HL-3 in this study may have the potential in aquaculture feed industry.

     

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