工业微生物2012，Vol.42Issue(1)：23-27,5.DOI:10.3969/j.issn.1001-6678.2012.01.005
BMMYA’平板-Fast blue RR顶层琼脂法高效筛选高酶活脂肪酶
BMMYA' plate -Fast blue RR top agar —— an efficient plate screening method for lipase with high enzyme activity
摘要
Abstract
An efficient plate screening method for lipase with high enzyme activity was established. This novel method was used as a powerful tool to screen a mutant library for strains with high enzyme activity or high yield of enzyme. Taking lipase gene proRCL from Rhizopus chinensis CCTCCM201021 as the template, the mutant pool was constructed in Pichia pastoris GS115 in earlier experiments. Candidate strains were inoculated into BMMYA' plates containing 2% (V/V) methanol, grew and induced for 4 ~ 5 d at 30 XI, afterward undergone heat-treatment at enzyme lethal temperature for 1 h, and then, cooled at ice and room temperature. Fast blue RR top agar was then poured into plates and positive strains demonstrated apparent dark brown around within 2 min. This method was very convenient, fast, efficient and accurate and with more than 90% screening credibility.关键词
Fast blue RR顶层琼脂法/BMMYA’平板/高温处理/脂肪酶平板筛选Key words
Fast blue RR top agar/ BMMYA' plate/ heat-treatment/ plate screening of lipase引用本文复制引用
王睿,喻晓蔚,沙冲,徐岩..BMMYA’平板-Fast blue RR顶层琼脂法高效筛选高酶活脂肪酶[J].工业微生物,2012,42(1):23-27,5.基金项目
项目名称:国家自然科学基金(No.20802027),国家高科技研究发展项目(863计划)(No.2010AA101501,No.2008AA102304),十一五支撑计划(No.2008BAI63B07),长江学者和创新团队发展计划(No.IRT0532),中央高校基本科研业务费专项资金(JUSRP11014). （No.20802027）
