基因组改组选育磷脂酶A1生产菌

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中国生物工程杂志 ›› 2013, Vol. 33 ›› Issue (10) : 59-66.

研究报告

基因组改组选育磷脂酶A1生产菌

王洲, 薛正莲, 马琦亚, 苏燕南, 赵世光

作者信息 +

Breeding of Phospholipase A1-producing Strains by Genome Shuffling

WANG Zhou, XUE Zheng-lian, MA Qi-ya, SU Yan-nan, ZHAO Shi-guang

Author information +

文章历史 +

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%。

Abstract

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%.

导出引用

王洲, 薛正莲, 马琦亚, 苏燕南, 赵世光. 基因组改组选育磷脂酶A1生产菌[J]. 中国生物工程杂志, 2013, 33(10): 59-66

WANG Zhou, XUE Zheng-lian, MA Qi-ya, SU Yan-nan, ZHAO Shi-guang. Breeding of Phospholipase A1-producing Strains by Genome Shuffling[J]. China Biotechnology, 2013, 33(10): 59-66

中图分类号： Q933

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