基因组改组选育磷脂酶A1生产菌  

Breeding of Phospholipase A1-producing Strains by Genome Shuffling

作　　者：王洲[1] 薛正莲[1] 马琦亚[1] 苏燕南[1] 赵世光[1] 

机构地区：[1]安徽工程大学生物与化学工程学院微生物发酵安徽省工程技术研究中心,芜湖241000

出　　处：《中国生物工程杂志》2013年第10期59-66,共8页China Biotechnology

基　　金：安徽省自然科学基金(11040606M81);安徽省高校自然科学基金(KJ2009A168)资助项目

摘　　要：以蜡样芽胞杆菌W22为出发菌株,通过紫外诱变和等离子体诱变得到若干株酶活力提高的突变株。以紫外诱变突变菌株W22-a、W22-g和等离子诱变菌株W22-5为基因组改组的亲本,考察了原生质体的制备、灭活条件。原生质体的最佳制备条件是:溶菌酶浓度0.5mg/ml,酶解时间90min,反应温度25℃,预处理甘氨酸浓度为20mg/ml。选择紫外灭活160秒、热灭活16分钟的剂量来完成亲本原生质体的灭活。从众多融合子中筛得一株突变株WR2-2,发酵酶活为17.78U/ml,相对于出发菌株W22(9.22U/ml)提高酶活达92.8%。Several mutant strains with the increased enzyme activity were gained when the original strain Bacillus cereus W22 was treated by UV irradiation and plasma. The mutants W22-a and W22-g obtained by irradiation and W22-5 got by plasma were used as the parental strains for genome shuffling, and the conditions of protoplast preparation and inactivated was studied. The optimal method of protoplast preparation was pretreated with glycin at the concentration of 20mg/ml, then treated with the lysozyme at the concentration of 0.5mg/ml, and the hydrolysis time was 90 min at reaction temperature 25℃. The UV treatment for 160sec and heat treatment for 16min were selected to complete the inactivation of the parental protoplasts. Mutant WR-2 has screened from many fusants, and the fermentation enzyme activity can be up to 17.78U/ml, compared to the enzyme activity of the original strain W22 which was 9.22U/ml, it increased 92.8%.

关 键 词：磷脂酶A1 紫外诱变 等离子诱变 基因组改组 

分 类 号：Q933[生物学—微生物学]