透明颤菌vgb基因的整合对生物法发酵香兰素的影响
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摘要
在以阿魏酸为底物生物法转化香兰素过程中,通过Red重组技术用外源的透明颤菌血红蛋白基因(vgb)取代大肠杆菌基因组icdA基因,使菌株可以适应高密度、低溶氧发酵,为香兰素的发酵生产提供新的尝试。CO差光谱分析显示整合的vgb基因表达产生了VHb,且该蛋白具有活性。发酵结果显示表达了vgb基因的新菌株与原重组菌株相比在低氧环境下仍能维持较高的生物量,且香兰素产量由原来的1 668 mg/l上升到2 240 mg/l,提高了约34. 2%。
In the process of biotransformation of vanillin using ferulic acid as substrate,the genomic icdA gene of Escherichia coli was replaced by foreign( vgb) gene of Vitreoscilla hemoglobin gene by Red recombination technology,so that the strain could adapt to high density and low dissolved oxygen fermentation. It provided a new attempt for the fermentation production of vanillin. CO differential spectroscopy analysis showed that the integrated vgb gene expression produced VHb,and the protein had activity. The results of fermentation showed that the new strain expressing vgb gene could maintain a higher biomass under hypoxic condition than the original recombinant strain,and the vanillin production increased by about 34. 2% from 1 668 mg/L to 2 240 mg/L.
In the process of biotransformation of vanillin using ferulic acid as substrate,the genomic icdA gene of Escherichia coli was replaced by foreign( vgb) gene of Vitreoscilla hemoglobin gene by Red recombination technology,so that the strain could adapt to high density and low dissolved oxygen fermentation. It provided a new attempt for the fermentation production of vanillin. CO differential spectroscopy analysis showed that the integrated vgb gene expression produced VHb,and the protein had activity. The results of fermentation showed that the new strain expressing vgb gene could maintain a higher biomass under hypoxic condition than the original recombinant strain,and the vanillin production increased by about 34. 2% from 1 668 mg/L to 2 240 mg/L.
引文
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[12] KHOSLA C,BAILEY J E. The Vitreoscilla hemoglobin gene:Molecular cloning,nucleotide sequence and genetic expression in Escherichia coli[J]. Mol Gen Genet,1988,214(1):158-161.
[13]周艳芬,赵晓瑜,张玉,等.透明颤菌血红蛋白基因在苏云金杆菌中的表达[J].河北大学学报(自然科学版),2006,26(1):33-37.
[2] MUHEIM A,MULLER B,MUNCH T,et al. Microbiological process for producing vanillin:US,US6235507[P]. 2001.
[3] HUA D,MA C,SONG L,et al. Enhanced vanillin production from ferulic acid using adsorbent resin[J]. Appl Microbiol Biotechnol,2007,74(4):783-790.
[4] RABENHORST J,HOPP R,BYNG G S. Process for the preparation of vanillin and suitable microorganisms:EP,EP0761817[P]. 2002.
[5]杨玉凤,费理文,李建华,等.重组大肠杆菌全细胞催化合成莱鲍迪苷D[J].工业微生物,2017,47(5):1-7.
[6] LEE E G,YOON S H,DAS A,et al. Directing vanillin production from ferulic acid by increased acetyl-CoA consumption in recombinant Escherichia coli[J]. Biotechnol Bioeng,2009,102(1):200-208.
[7] AYDIN S,WEBSTER D,STARK B C,et al. Nitrite inhibition of Vitreoscilla hemoglobin(VHb)in recombinant E. coli:direct evidence that VHb enhances recombinant protein production[J].Biotechnol Prog,2000,16(6):917-921.
[8]于慧敏,史悦,沈忠耀,等. CO差光谱法分析重组大肠杆菌中的透明颤菌血红蛋白[J].清华大学学报(自然科学版),2002,(05):615-618.
[9]阎博,赵林,谭欣,等.浊度法快速测定培养液中的生物量[J].天津化工,2003,17(1):45-46.
[10] YOON S H,LI C,LEE Y M,et al. Production of vanillin from ferulic acid using recombinant strains of Escherichia coli[J]. Biotechnol Bioproc E,2005,10(4):378-384.
[11] MASAI E,HARADA K,PENG X,et al. Cloning and characterization of the ferulic acid catabolic genes of Sphingomonas paucimobilis SYK-6[J]. Appl Environ Microbiol,2002,68(9):4416-4424.
[12] KHOSLA C,BAILEY J E. The Vitreoscilla hemoglobin gene:Molecular cloning,nucleotide sequence and genetic expression in Escherichia coli[J]. Mol Gen Genet,1988,214(1):158-161.
[13]周艳芬,赵晓瑜,张玉,等.透明颤菌血红蛋白基因在苏云金杆菌中的表达[J].河北大学学报(自然科学版),2006,26(1):33-37.