等离子体作用结合氧限制模型选育辅酶Q_(10)高产菌株  

Screening of High-Yield Coenzyme Q_(10) Producing Strain by Using Atmospheric and Room Temperature Plasma and Oxygen-Limited Model

作　　者：张幸子 王晓惠 王泽建[1] 陈必钦 李丹 郭美锦[1] 储炬[1] 庄英萍[1] ZHANG Xingzi;WANG Xiaohui;WANG Zejian;CHEN Biqin;LI Dan;GUO Meijin;CHU Ju;ZHUANG Yingping(State Key Laboratory of Bioreactor Engineering,National Engineering Research Center for Biotechnology(Shanghai),East China University of Science and Technology,Shanghai 200237,China;Inner Mongolia Kingdomway Pharmaceutical Company,Huhhot 010000,China)

机构地区：[1]华东理工大学生物反应器工程国家重点实验室,国家生化工程技术研究中心(上海),上海200237 [2]内蒙古金达威药业有限公司,呼和浩特010000

出　　处：《华东理工大学学报（自然科学版）》2021年第3期308-315,共8页Journal of East China University of Science and Technology

基　　金：国家重点研发计划项目(2017YFF0204602)。

摘　　要：将常温常压等离子体(Atmospheric and Room Temperature Plasma,ARTP)诱变和无水亚硫酸钠构建的氧限制筛选模型相结合,通过适合辅酶Q_(10)生产菌株的24孔板高通量快速培养技术,选育了类球红细菌Rhodobacter sphaeroides耐氧限的高性能突变株。实验结果表明:该菌株诱变筛选最合适的ARTP诱变时间为25 s,氧限制平板上无水亚硫酸钠的最佳质量浓度为0.4 g/L,24孔板上该菌氧传递的最优装液量为2.0 mL、发酵周期为48 h,在上述条件下,最终选育出了氧限制条件下辅酶Q10合成效率高的突变菌株R.sphaeroides F5D13。在5 L发酵罐分批培养过程中,该高产菌株表现出了较强的氧亲和力,单位菌体的合成效率比出发菌株提升了18%。Coenzyme Q_(10) is an important hydrogen transmitter in respiratory chain,which plays an important role in the human body.It has been widely used for the treatment of cardiovascular and cerebrovascular diseases,and has been applied in cosmetics,health care products and other aspects.With the development of research,the market demand and industrial output of coenzyme Q_(10) are constantly expanding.Microbial fermentation is the most promising method for the production of coenzyme Q10.This study used atmospheric and room temperature plasma(ARTP)towards Rhodobacter sphaeroides to obtain mutants.Meanwhile,an oxygen-limited model was established to screen the strains which could suffer the low oxygen concentration.The bacterial suspension was treated by ARTP for 25 s,then cultured on the plate with 0.4 g/L sodium sulfite.The strains were fermented in 2.0 mL volume for 48 h.In the primary screening 24-well plates,6 mutants were obtained.Furthermore,these mutants were cultured in shake flask to verify genetic stability.A mutant R.sphaeroides F5D13 presented good stability and production of Coenzyme Q10.The titer of R.sphaeroides F5D13 was improved from 86.2 mg/L to 111.8 mg/L compared to the original strain.Finally,the yield of Coenzyme Q10 reached 770.06 mg/L in 5 L fermentation after 100 h,which was increased by 18.0%.According to the fermentation parameters,the mutant R.sphaeroides F5D13 showed the advantage of biomass in the cell growth phase and strong oxygen demand in product synthesis phase,which leaded to the higher oxygen uptake rate and yield than the original parent strain.

关 键 词：常温常压等离子体 氧限制模型 辅酶Q_(10) 高通量筛选 类球红细菌 

分 类 号：TS201.3[轻工技术与工程—食品科学]