柴油降解酵母复合菌的筛选、鉴定及培养条件优化  

Screening and Identification of Diesel-degrading Composite Yeasts and Optimal Study on Cluture Conditions

作　　者：陈梦楠[1] 颜贻晨[1] 刘倩倩[1] 张莉[1] 马云婷[1] 袁双双 刘雪珠[1] 

机构地区：[1]浙江海洋学院海洋科学与技术学院,浙江舟山316000

出　　处：《湖北农业科学》2016年第2期318-322,共5页Hubei Agricultural Sciences

基　　金：国家级大学生创新训练项目(201310340001)

摘　　要：以柴油为惟一碳源,用富集、驯化培养基从舟山港口柴油污染海水和海泥中筛选得到4株高效柴油降解酵母菌(Yeast)C_1、C_2、D_1、D_3,其柴油降解率为(70.49±2.50)%^(78.76±1.24)%。对4株菌进行3菌和4菌的相同比例的随机组合构建混合菌群,从中筛选得到最佳柴油降解复合酵母菌S1-5(C_2+D_1+D_3),降解率达到(85.30±1.29)%。再通过形态学观察、生理生化特征分析及16S rDNA、26S rDNA测序等方法对构成最佳降解复合菌的3株单菌进行鉴定。通过分子鉴定,得到酵母菌C_2、D_1、D_3都是耶罗维亚酵母属(Yarrowia),其中C_2和D_3是解脂耶罗维亚酵母(Yarrowia lipolytica)。在模拟污染水域实际物化条件的基础上优化培养条件,优化试验获得酵母复合菌S1-5的最佳培养条件为盐度30‰,NH_4NO_31.5 g/L,MgSO_4·7H_2O 0.3 g/L,KCl 1 g/L,FeCl_3·6H_2O 50 mg/L,在该条件下其降解率达到(82.79±1.38)%。Four efficient diesel-degradation yeast strains named C1,C2,D1 and D3 were screened from being polluted seawater and mud in zhoushan port after enrichment and domestication, and diesel was sole carbon source in the progress. The diesel degradation rate of around（70.5±2.50）% to（78.8±1.24）%. The selected yeast strains were made into ten combinations respectively, as three or four strains were randomly mixed. The results indicated that S1-5（C2＋D1＋D3） showed optimal degradation ability at（85.30±1.29）%. The composite yeast of three single yeast strains were identified through the morphological observation of physiological, biochemical characteristics analysis 16 S rDNA and 26 S rDNA sequencing methods. By the molecular identification, the result showed that the strains C2,D1 and D3 were Yarrowia, among them the strains C2 and D3 were Yarrowia lipolytica. According to single factor experiment and orthogonal experiment, the optimal degradation culturing condition for yeast strain group S1-5 were initially established. Under the optimized condition of NaCl 30 g / L, NH4NO31.5 g / L,MgSO4·7H2O 0.3 / L, KCl 1 g / L, FeCl3·6H2O 50 mg / L,the degradation rate gained the highest one, that was（82.79±1.38）%.

关 键 词：酵母菌(Yeast) 柴油降解 筛选 鉴定 优化 

分 类 号：X172[环境科学与工程—环境科学]