原生质体激光诱变选育少根根霉脂肪酶高产菌株  

Breeding of Rhizopus arrhizus lipase producing strain by laser induced protoplast mutagenesis

作　　者：冀颐之[1,2] 李政[3] 谭天伟[4] 杜连祥[1] 林强[2] 

机构地区：[1]天津科技大学生物工程学院,天津市工业微生物重点实验室,天津300222 [2]北京联合大学生物化学工程学院,北京100023 [3]天津工业大学纺织学院,天津300160 [4]北京化工大学教育部生物炼制工程中心,北京100029

出　　处：《工业微生物》2011年第3期38-43,共6页Industrial Microbiology

基　　金：国家"863"项目(No.2002AA514030);国家"十五"攻关项目(No.2001BA708B02208);国家自然科学基金项目(No.2107600C);中石化项目(No.202059)

摘　　要：对少根根霉BUCT-11原生质体制备、再生条件及激光诱变育种进行了研究。结果显示,少根根霉BUCT-11原生质体形成及再生最佳条件为:菌龄24 h,混合酶由27mg/mL的蜗牛酶和53 mg/mL的纤维素酶组成,酶解时间1.5 h,酶解温度30℃,渗透压稳定剂为0.6 mol/L NH_4 Cl、0.02 mol/L MgSO_4·7H_2O、0.4 mol/L CaCl_2的磷酸盐缓冲液。所得原生质体经激光诱变选育获得具有稳定遗传特性的高产突变菌株Y007,在摇瓶发酵条件下,测得脂肪酶水解酶活为151 U/mL,较出发菌株酶活提高了41%。The conditions of protoplast formation, regeneration and laser mutagenesis of Rhizopus arrhizus BUCT-11 strain were investegated. As a result, the optimum conditions for protoplast preparation and regeneration of Rhizopus arrhizus BUCT-11 strain were as follows： 24 h of mycelium growth time, compound enzyme containing 27 mg/mL of snailase and 53 mg/mL of fibrin enzyme, 1.5 h hydrolysis, hydrolysis temperature at 30℃, applying 0.6 mol/L NH4 Cl, 0.02 mol/L MgSO4·7H2O, 0.4 mol/L CaC12 phosphate buffer for osmotic stabilizer. The He-Ne laser was irradiated on protoplasts of strain BUCT-11. The high-yielding lipase mutant numbered Y007 with good genetic stability was obtained, and its lipase yield in shaking flask scale was up to 151 U/mL, which was 41% higher than that of the parent strain.

关 键 词：脂肪酶 少根根霉 激光 原生质体 诱变选育 

分 类 号：Q93[生物学—微生物学]