多目标优化新型除草剂高粱素生物合成关键酶ARS的异源双控表达

Multi-objective Optimization of Dual Control Heterologous Expression of Sorghum bicolor ARS for Novel Sorgoleone Herbicide Biosynthesis

[目的]基于试验设计对高粱素生物合成关键酶-烷基间苯二酚合酶(Sb_ARS)的异源双控表达进行多目标优化。[方法]在前期成功进行Sb_ARS的温控自裂解异源表达的基础上,依次通过PB设计、最陡爬坡设计、响应面设计及线性加权多目标优化等方法,分析了11个与Sb_ARS在大肠杆菌中双控异源表达密切相关因素的影响。[结果]菌体培养时长、IPTG诱导时长和甘氨酸浓度对Sb_ARS酶活力和裂解率影响显著;当菌体培养时长为6.8h、IPTG诱导时长为4.6 h和甘氨酸质量浓度为1.0 g/L时,可获得总体最优Sb_ARS酶活力(117.12±0.13)%和裂解率(89.21±0.35)%。[结论]研究结果为新型除草剂高粱素的生物合成研究奠定理论基础。

[Aims] Statistical methodology was applied to optimize the heterologous double-regulated expression of alkylresorcinol synthase(Sb_ARS), a key enzyme for sorgoleone herbicide biosynthesis. [Methods] Plackett-burman design, path of steepest ascent, response surface methodology and multi-objective optimization was used to analyzed 11 factors, which were related to the heat-inducible autolytic heterologous expression of Sb_ARS in E. coli. [Results] Cell growth time, IPTG induction time and glycine concentration were found to have significant effects on Sb_ARS activity and lysis efficiency. The optimum operation conditions was cell growth time 6.8 h, IPTG induction time 4.6 h and glycine concentration 1.0 g/L, under which Sb_ARS activity of(117.12±1.03)% and lysis efficiency of(89.21±0.71)%were simultaneously obtained. [Conclusions] The results would lay theoretical basis for the biosynthesis of new herbicide sorghum.