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耐甲醇脂肪酶产生菌的筛选鉴定及其酶学性质研究
引用本文:王斌,宗浩,张志敏,王琰,万一.耐甲醇脂肪酶产生菌的筛选鉴定及其酶学性质研究[J].安徽农业科学,2011,39(20):12001-12004.
作者姓名:王斌  宗浩  张志敏  王琰  万一
作者单位:1. 四川师范大学生命科学院,四川,成都,610068
2. 陕西省微生物研究所,陕西,西安,710043
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。

关 键 词:脂肪酶  菌种筛选  鉴定  耐甲醇  酶学性质

Screening of Methanol-tolerant Lipase-producing Strains and Study of Properties of Lipase
Institution:WANG Bin et al(College of Life Science,Sichuan Normal University,Chengdu,Sichuan 610068)
Abstract: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.
Keywords:Lipase  Strain isolation  Identification  Methanol-tolerant  Properties of lipase
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