Biosynthesis of phycocyanobilin in recombinant Escherichia coli  

作　　者：MA Chengbo LI Wenjun GE Baosheng LIN Jian QIN Song 

机构地区：[1]College of Life Sciences,Yantai University,Yantai 264005,China [2]Yantai Institute of Coastal Zone Research,Chinese Academy of Sciences,Yantai 264003,China [3]Center for Bioengineering and Biotechnology,China University of Petroleum(East China),Qingdao 266580,China [4]Center for Ocean Mega-Science,Chinese Academy of Sciences,Qingdao 266071,China

出　　处：《Journal of Oceanology and Limnology》2020年第2期529-538,共10页海洋湖沼学报（英文）

基　　金：Supported by the National Natural Science Foundation of China(No.41906109);the Key Research and Development Program of Yantai(No.Y933021011)

摘　　要：The recombinant expression of phycocyanobilin(PCB)was carried out in Escherichia coli,and the best fermentation conditions of recombinant E.coli biosynthesized PCB are optimized in the response surface methodology to improve PCB production.The recombinant PCB is extracted,isolated,and purified by methanol and chloroform extraction.Recombinant PCB is validated in UV-vis spectroscopy,high-pressure liquid chromatography,and mass spectrometry.In addition,the anti-oxidant activities of the recombinant PCB are determined.The best induction conditions that optimized by Design Expert 8.0 software include:lactose concentration 4 mmol/L,induction temperature 24.69℃,induction time 4.6 h,and induction duration 13.57 h,under which the PCB expression level reached approximately 13 mg PCB/L,which is more than four times of previously reported 3 mg PCB/L.The maximum absorption peak of the recombinant PCB is located at 680 nm with a high fluorescence intensity of 470 nm.The recombinant PCB has a good ability to scavenge 2,2-diphenyl-1-picrylhydrazyl(DPPH)free radicals.The recombinant expression of phycocyanobilin(PCB) was carried out in Escherichia coli,and the best fermentation conditions of recombinant E.coli biosynthesized PCB are optimized in the response surface methodology to improve PCB production.The recombinant PCB is extracted,isolated,and purified by methanol and chloroform extraction.Recombinant PCB is validated in UV-vis spectroscopy,high-pressure liquid chromatography,and mass spectrometry.In addition,the anti-oxidant activities of the recombinant PCB are determined.The best induction conditions that optimized by Design Expert 8.0 software include:lactose concentration 4 mmol/L,induction temperature 24.69℃,induction time 4.6 h,and induction duration 13.57 h,under which the PCB expression level reached approximately 13 mg PCB/L,which is more than four times of previously reported 3 mg PCB/L.The maximum absorption peak of the recombinant PCB is located at 680 nm with a high fluorescence intensity of 470 nm.The recombinant PCB has a good ability to scavenge 2,2-diphenyl-1-picrylhydrazyl(DPPH) free radicals.

关 键 词：combinational biosynthesis PHYCOCYANOBILIN response surface methodology ANTIOXIDANT 

分 类 号：Q936[生物学—微生物学]