生物加工过程2018，Vol.16Issue(1)：82-88,7.DOI:10.3969/j.issn.1672-3678.2018.01.010
一种来源于酒曲地衣芽孢杆菌溶菌酶基因在毕赤酵母中的诱导表达及活性分析
Cloning,expression and characterization of a lysozyme gene of Bacillus licheniformis isolated from fermented grains
摘要
Abstract
We used the gradient dilution and streak plate methods to isolate and purify microorganisms from fermented grains.The microorganisms were cultured in liquid medium and the lysozyme activity were detected. Through observation of the morphological characteristics and molecular biological characterization,the bacterial strain with lysozyme activity was identified as Bacillus licheniformis. After extracting the bacteria genome DNA and designing primer,the lysozyme gene Lyz was cloned and inserted into the inducible yeast expression vector pPICZαA, constructed the recombinant expression plasmid pPICZαA-Lyz and to be transformed into Pichia pastoris X33.Shake flask fermentation indicated that the activity of the lysozyme of multi-copy transformant reached 1360 U/mL after cultivation in buffered methanol-complex medium( BMMY) for 72 h. The lysozyme was very stable in a broad pH range ( 3. 0-9. 0) and thermostable. The antimicrobial activity of lysozyme showed that it had antimicrobial activity against several gram-positive strains.关键词
地衣芽孢杆菌/溶菌酶/毕赤酵母/重组表达Key words
Bacillus licheniformis/lysozyme/Pichia pastoris/expression分类
生物科学引用本文复制引用
王凤寰,韩煦,孙博,赵飞函,温赛,杜鉴,廖永红..一种来源于酒曲地衣芽孢杆菌溶菌酶基因在毕赤酵母中的诱导表达及活性分析[J].生物加工过程,2018,16(1):82-88,7.基金项目
国家重点研发计划(2017YFB0308900) （2017YFB0308900）
国家自然科学基金(21406005) （21406005）
中国轻工业清洁生产和资源综合利用重点实验室开放课题 （）
科技创新服务能力建设——轻工技术与工程学科发酵工程研究平台( PXM2017_01423_000034) （ PXM2017_01423_000034）
