轻工学报2026，Vol.41Issue(1)：69-80,12.DOI:10.12187/2026.01.007
异源表达透明颤菌血红蛋白结合调控策略提高全细胞催化合成苯乳酸产量的研究
Study on improving the yield of whole-cell catalytic production of phenyllactic acid by heterologous expression of Vitreoscilla hemoglobin combined with regulation strategy
摘要
Abstract
[Objective]This study aims to improve the oxygen(O2)utilization rate of strains in the whole-cell catalytic reaction system and efficiently catalyze the synthesis of phenyllactic acid(PLA)from phenylalanine(PHE).[Methods]Based on the effect of heterologous expression of Vitreoscilla hemoglobin(VHb)on the growth of host Escherichia coli,genetic engineering technology was used to construct a multi-enzyme coupled expression system including VHb,L-amino acid deaminase(L-AAD)and lactate dehydrogenase(LDH).Subsequently,the expression regulation strategies of key enzymes were optimized,and the genetic stability of recombinant strains was evaluated by continuous passage experiments.[Results]Heterologous expression of VHb could improve the oxygen utilization rate of strains and significantly promote the growth rate and maximum biomass of host E.coli.In shake flask culture,the time for the strain to enter the logarithmic growth phase during the scale-up culture stage was shortened by 1.5 h,and the maximum biomass was increased by 46%when VHb expression was induced alone.The dominant recombinant strain E.coli BL21(pRSFDuet-aad-ldh-T7-rbs2-2vhb)was obtained by optimizing the VHb expression regulation strategy.Furthermore,the whole-cell catalytic reaction system was optimized,and the optimal reaction conditions were determined as follows:0.010 mol/L PBS buffer,pH 7.0,temperature 37℃,and rotation speed 250 r/min.Under these conditions,the maximum PLA yield of the dominant recombinant strain was 53.7 g/L;The dominant recombinant strain had good stability;after 10 consecutive passages,the change rate of PLA yield in each generation was less than 5%,showing potential to become an industrial strain.[Conclusion]Heterologous co-expression of VHb can promote the growth of E.coli and improve the yield of PLA via whole-cell catalysis.关键词
多酶偶联/全细胞催化/透明颤菌血红蛋白/苯乳酸/重组菌株Key words
multi-enzyme conjugation/whole-cell catalysis/Vitreoscilla hemoglobin/phenyllactic acid/recombinant strain分类
轻工纺织引用本文复制引用
丁心成,孙瑞杰,贾士儒,侯颖..异源表达透明颤菌血红蛋白结合调控策略提高全细胞催化合成苯乳酸产量的研究[J].轻工学报,2026,41(1):69-80,12.基金项目
工业发酵微生物教育部重点实验室暨天津市工业微生物重点实验室资助课题项目(2020KF006) （2020KF006）
天津市研究生科研创新项目(2022SKYZ092) （2022SKYZ092）
