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当前位置: 首页 > 文章 > 耐甲醇脂肪酶产生菌的筛选鉴定及其酶学性质研究 安徽农业科学 2011,39 (20) 12001-12004
Position: Home > Articles > Screening of Methanol-tolerant Lipase-producing Strains and Study of Properties of Lipase Journal of Anhui Agricultural Sciences 2011,39 (20) 12001-12004

耐甲醇脂肪酶产生菌的筛选鉴定及其酶学性质研究

作  者：
王斌;宗浩;张志敏;王琰;万一
单  位：
四川师范大学生命科学院;陕西省微生物研究所
关键词：
脂肪酶;菌种筛选;鉴定;耐甲醇;酶学性质
摘  要：
[目的]筛选耐甲醇脂肪酶产生菌,并对其酶学性质进行研究。[方法]采用培养基中添加不同浓度甲醇(0、5%、10%、15%、20%)的方法筛选甲醇耐受性脂肪酶产生菌,形态学和ITS序列分析确定菌株的种属关系,硫酸铵分级沉淀和DEAE-Sepharose FF纯化脂肪酶,并对纯化的酶进行耐甲醇试验和酶学性质研究。[结果]筛选到183株脂肪酶产生菌,菌株SXL-107对浓度20%甲醇具有较好耐受性,脂肪酶活力达104 U/ml;ITS同源性分析确定该菌为Galactomyces geotrichum;经过纯化的SXL-107脂肪酶在浓度10%甲醇溶液中作用48 h后,剩余酶活为96.15%;该酶最适温度为40℃,pH为7.0,50℃温浴60 min仍有80%的酶活力,在pH 4.0~9.0间稳定,微量的Mg2+、K+和Zn2+对该酶有激活作用,Mn2+和CO2+有抑制作用,分子量约为66 kDa。[结论]筛选到一株耐甲醇能力较强的脂肪酶产生菌,为该酶在生物柴油中的应用奠定基础。
译  名：
Screening of Methanol-tolerant Lipase-producing Strains and Study of Properties of Lipase
作  者：
WANG Bin et al(College of Life Science,Sichuan Normal University,Chengdu,Sichuan 610068)
关键词：
Lipase;Strain isolation;Identification;Methanol-tolerant;Properties of lipase
摘  要：
[Objective] The methanol-tolerant lipase-producing strains was screened and the properties of lipase was studied.[Method] Methanol-tolerant lipase-producing strains were screened with the method to add different concentrations of methanol(0.5%,10%,15%,20%) to the medium.SXL-107 was identified as Galactomyces geotrichum by its morphological and ITS sequences.The lipase was depurated by ammonium sulfate fractionation and DEAE-Sepharose FF.Properties of lipase and methanol-tolerance of purified enzyme were studied.[Result] 183 lipase-producing strains were isolated from rich oil soil.The strain SXL-107 had a better tolerance to 20% methanol.The enzyme activity of SXL-107 achieved 104 U/ml.It was identified as Galactomyces geotrichum by its morphological and ITS sequences.When SXL-107 lipase reacted in 10% methanol for 48 h,the remaining activity was 96.15%.The optimal pH was 7.0 and the temperature of the lipase was 40 ℃.The activity of the lipase reached 80% at 50 ℃ for 60 min,and was stable from pH 4.0 to 9.0.The enzyme activity was stimulated by Mg2+,K+and Zn2+,but inhibited by Mn2+ and CO2+.The molecular weight of the purified SXL-107 lipase was about 66 kDa.[Conclusion] A Methanol-tolerant lipase-producing strain was screened and it provided the foundation for bio-diesel production.

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