重组亚胺还原酶工程菌快速高密度发酵研究

Rapid high-density fermentation study on Escherichia coli recombinant strain for expressing imine reductase

通过快速高密度发酵培养,以重组亚胺还原酶大肠杆菌工程菌的菌体生物量及酶活力作为评价标准,利用低成本的发酵培养基,在短时间内获得菌体的最大生物量和最佳酶活力。采用2 L发酵体系,对该工程菌的接种量和诱导条件进行优化。结果表明,该菌株的快速高密度发酵最佳接种量为10%,当生物量OD_(600)值达到12时,发酵体系降温至20℃,加入0.4 mmol/L异丙基-β-D-硫代半乳糖苷,为最佳诱导条件,并以此条件进行20 L快速高密度发酵,诱导12 h酶活力最高,为4.56 U/g。该研究为进一步放大发酵体系以实现亚胺还原酶工业化快速高产量制备的生产奠定基础。

Rapid high-density fermentation was employed to obtain maximal biomass and enzyme activity of recombinant imine reductase-expressing Escherichia coli engineered strain using a low-cost fermentation medium as the culture medium.The bacterial biomass and enzyme activity were used as evaluation indicators.A 2 L fermentation system was used to optimize the inoculation amount and induction conditions for the engineered strain.The results showed that the optimal inoculation amount for the strain was 10%,and when the biomass OD_(600)reached 12,the fermentation system was cooled to 20℃,and 0.4 mmol/L isopropyl-beta-D-thiogalactoside(IPTG)was added as the best induction condition.Under this condition,a 20 L rapid high-density fermentation was performed,and the maximum enzyme activity was 4.56 U/g after induction for 12 h.This study lays the foundation for further scaling up the fermentation system to achieve rapid and high-yield production of imine reductase for industrial applications.