犹他游动放线菌中酰胺水解酶基因拷贝数增加对转化棘白菌素B效率的影响

摘要

Objective:To construct genetic engineering strain containing more copies of echinocandin B deacylase by φ31-directed site-specific recombination. Methods:The gene integrated plasmid pYGCQ-03-13 containing echinocandin B deacylase gene was constructed and transformed into Actinoplanes utahensis strain SIPI-T2001 with conjugation. The bioconversion efficiency of the recombination strain was studied with method of resting cells conversion. Results; A recombinant strain named SIPI-GE. T2001 was screened and its bioconversion efficiency was increased to 61. 7% . Conclusion; Increasing the copies of echinocandin B deacylase gene in Actinoplanes utahensis can improve the bioconversion efficiency for echinocandin B.%目的:利用ΦC31整合酶介导的位点特异性重组方法,构建棘白菌素B酰胺水解酶基因拷贝数增加的犹他游动放线菌重组菌株.方法:构建含有棘白菌素B酰胺水解酶基因的基因整合型质粒pYGCQ-03-13,通过属间接合转移的方法将该基因片段转入犹他游动放线菌SIPI-T2001中；利用静息细胞转化法,考察其对转化效率的影响.结果:筛选得到的阳性重组菌株SIPI-GE.T2001,对棘白菌素B的转化效率最高达到61.7％.结论:增加棘白菌素B酰胺水解酶基因拷贝数,有效地提高了犹他游动放线菌对棘白菌素B的转化效率.