一株饲用产脂肪酶芽孢杆菌的筛选及其紫外诱变育种  

Screening and ultraviolet mutation breeding of a forage Bacillus subtilis producing lipase

作　　者：郭晓军[1] 郭威[1] 袁洪水[1] 朱宝成[1] 

机构地区：[1]河北农业大学生命科学学院,河北保定071000

出　　处：《中国饲料》2015年第12期27-29,共3页China Feed

基　　金：国家星火计划项目(2010GA105010);河北省科技计划项目(12236606)

摘　　要：本试验采用溴甲酚紫显色法和三丁酸甘油酯平板法相结合的方法 ,从富含油脂的土样中筛选产脂肪酶的芽孢杆菌菌株。采用橄榄油乳化法对溶解圈较大的菌株进行酶活力的测定,得到一株酶活力较高的菌株3EA1-1,酶活力达到36.1 U/m L。结合该菌株的菌落形态、菌体特征、生理生化试验结果和16S r DNA序列分析结果,鉴定该菌株为枯草芽孢杆菌(Bacillus subtilis)。对其进行紫外线诱变,通过自然分离纯化,选育出了一株稳定高产菌株,命名为EA-30-A,其酶活力较原菌株提高了40%。经5次传代试验,酶活力较传代前降低2.27%(P>0.05),证明菌株的遗传稳定性良好。In this study,bromcresol purple method and tributyrin plate method were used to screen the Bacillus strains producing lipase from greasy soil samples.Olive oil emulsion method was used to measure the enzyme activity of the strains. Finally, strain 3EA 1-1 was chosen with lipase activity of 36.1 U/mL.According to colony morphological,mycelia physiological,biochemical characteristics and 16S rDNA sequences analysis results,strain 3EA1-1 was identified as Bacillus subtilis. After ultraviolet mutation and the natural process of separation and purification,the strain named EA-30-A was selected as the highest lipase producing strain,lipase activity of which was increased by 40%.The strain EA-30-A was proved a good genetic stabihty strain by plate cuhure of five generations ,activity of which decreased by 2.27% than before.

关 键 词：脂肪酶 芽孢杆菌 筛选 鉴定 紫外诱变 

分 类 号：S816.3[农业科学—饲料科学]