摘要
[目的]获得产酶能力强、环境安全友好的产脂肪酶菌株。[方法]采用罗丹明B平板法,从受油污污染土壤和水体中筛选高产脂肪酶菌株;利用血平板接种及抗生素药敏试验确定菌株安全性;通过氮源、碳源及诱导剂单因素实验结合响应曲面法优化菌株的发酵产酶条件;对菌株所产的脂肪酶进行酶学特性分析。[结果]从受油污污染土壤中分离筛选出一株具有高产脂肪酶能力的菌株JC-7,经鉴定为沙雷氏菌(Serratia sp.)。菌株于血琼脂平板上呈γ-溶血,对多数抗生素敏感。响应曲面法优化后酶活力最高达38.6μmol/min。菌株JC-7所产脂肪酶属于中温碱性脂肪酶。Ca^(2+)和Mg^(2+)能提高该酶的酶活力。该酶在丙三醇、二甲基亚砜以及甲醇中能维持较高的酶活力。十二烷基硫酸钠(SDS)、十六烷基三甲基溴化铵(CTAB)和TritonX-100都对该酶有激活作用,二硫苏糖醇(DTT)对该酶的酶活力几乎无影响,乙二胺四乙酸(EDTA)则抑制了该酶的酶活力。[结论]分离所获沙雷氏菌JC-7产脂肪酶能力强,对环境安全友好,优化培养条件后产酶能力明显提高,且该酶能适应较为广泛的作用条件,可为脂肪酶制剂的开发和环境污染生物降解菌剂的研究提供支持。
[Objective]Obtain a lipase-producing strain with strong enzyme production ability and environmental safety.[Method]The rhodamine B plate method was used to screen high-lipase-producing strains from oil-contaminated soil and water.The safety of the strains was determined by blood plate inoculation and antibiotic sensitivity test.The fermenta-tion enzyme production conditions of the strains were optimized by nitrogen source,carbon source and inducer single fac-tor experiment combined with response surface methodology.The enzymatic characteristics of the lipase produced by the strains were analyzed.[Result]A strain JC-7 with high lipase production ability was isolated and screened from oil-contaminated soil.It was identified as Serratia sp.The strain wasγ-hemolytic on blood agar plate and sensitive to most antibiotics.After optimization by response surface methodology,the enzyme activity reached up to 38.6μmol/min.The li-pase produced by strain JC-7 is a mesophilic alkaline lipase.Ca^(2+)and Mg^(2+)can improve the enzyme activity.The enzyme can maintain high activity in glycerol,dimethyl sulfoxide and methanol.Sodium dodecyl sulfate(SDS),hexadecyl tri-methyl ammonium bromide(CTAB)and TritonX-100 all activated the enzyme,dithiothreitol(DTT)had almost no effect on the enzyme activity,and ethylene diamine tetraacetic acid(EDTA)inhibited the enzyme activity.[Conclusion]The isolated Serratia JC-7 has a strong ability to produce lipase,is safe and friendly to the environment,and the enzyme pro-duction capacity is significantly improved after optimizing the culture conditions.The enzyme can adapt to a wider range of action conditions,which can provide support for the development of lipase preparations and the research on environ-mental pollution biodegradation agents.
作者
陈梦越
孙劲
张文丽
王洁
周星朴
黄诗维
楚菲菲
CHEN Mengyue;SUN Jin;ZHANG Wenli;WANG Jie;ZHOU Xingpu;HUANG Shiwei;CHU Feifei(School of Resources and Environment,Xichang University,Xichang 615013,Sichuan,China;School of Animal Science,Xichang University,Xichang 615013,Sichuan,China)
出处
《西昌学院学报（自然科学版）》
2024年第3期78-89,共12页
Journal of Xichang University(Natural Science Edition)
基金
凉山州科技计划项目(21ZDF0181)
西昌学院“两高”人才科研支持计划项目(LGZ202001)
国家大学生创新计划项目(202210628023)
四川省大学生创新训练项目(S202310628086)。
关键词
脂肪酶
响应曲面
酶学性质
lipase
response surface
enzyme properties
作者简介
陈梦越(2001-),男,四川甘孜人,本科生,研究方向:环境科学,e-mail:1967609164@qq.com。;通信作者:孙劲(1975-),男,四川邛崃人,副教授,硕士,研究方向:环境微生物及基于微生物发酵的固废资源化处理,e-mai:sunjin_xcc@xcc.edu.cn。
