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

Influence of amidohydrolase gene copy number increasing in Actinoplanes utahensis strain on echinocandin B bioconversion efficiency

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。

Objective： To construct genetic engineering strain containing more copies of echinocandin B deacylase by ФC31-directed site-specific recombination. Methods：The gene integrated plasmid pYGCQ-03-13 containing echinocandin B deaeylase 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 bioeonversion 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.