Trichoderma lentiforme ACCC30425脂肪酶基因的原核表达及其酶学性质的初步研究

Prokaryotic Expression of Recombinant Lipases Gene from Trichoderma lentiforme ACCC30425 and Preliminary Study of its Enzymatic Properties

基于在Trichoderma lentiforme ACCC30425全基因组序列注释的15个脂肪酶基因,进行脂肪酶基因合成与Escherichia coli BL21异源表达,获得11个产脂肪酶阳性克隆,其中酶活力最高为1.5 U/mL。经初步酶学分析,结果表明重组表达脂肪酶普遍具有耐热性、耐碱性的特征,最适温度范围为50~90℃,最适pH范围为8.5~9.5。其中,LipP最适温度为90℃,远高于目前已知脂肪酶的最适温度; LipQ最适温度为50℃,最接近洗涤工业的中温温度;其他脂肪酶最适温度为70℃或80℃,均属耐热脂肪酶。

According to the genome analysis of Trichoderma lentiforme ACCC30425,the coding sequences of 15 lipase genes were annotated,and then used for gene synthesis and heterologous expression in Escherichia coli BL21.11 lipase-producing clones were obtained,and the highest lipase activity was up to 1.5 U/mL.Results of preliminary enzymatic analysis indicated that the recombinant lipases had the characteristics of heat resistance and alkali resistance universally,showing optimal activities at 50~90℃and pH 8.5~9.5.Among them,the optimum temperature of LipP was 90℃,which was much higher than all known counterparts,and LipQ showed optimal activity at 50℃,which was close to the intermediate temperature of detergent industry.And other lipases were also thermostable lipase,showing optimal activities at 70℃or 80℃.