重组大肠杆菌转化富马酸生产L-丙氨酸
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摘要
通过克隆来源于睾丸酮丛毛单胞菌的L-天冬氨酸-β-脱羧酶基因,在一株具有L-天冬氨酸酶生产能力的大肠埃希氏菌CICC 11022S中异源表达,构建转化富马酸生产L-丙氨酸的重组工程菌。结果发现:重组工程菌9 h转化富马酸产生112.7 g/L的L-丙氨酸,生产速率12.5 g/(L·h),转化率93.8%。富马酸价格较低,有效降低L-丙氨酸生产的原料成本。通过构建重组工程菌,以富马酸为底物,高效生产L-丙氨酸,结果表明该方法具有较好的工业应用潜力。
The gene coding for L-aspartate-β-decarboxylase from Comamonas testosteroni was cloned and expressed in an Escherichia coli strain CICC 11022 S with the capability for L-aspartic acid production.The recombinant strain was used for the synthesis of L-alanine. The recombinant strain produced 112. 7g / L L-alanine from fumaric acid with a rate of 12. 5 g /( L·h) and a conversion ratio of 93. 8%. Fumaric acid has a low price,which efficiently reduces the substrate cost for L-alanine production. L-alanine was producd from fumaric acid by using the recombinant strain,and the results showed that the technology had industrial application potential.
The gene coding for L-aspartate-β-decarboxylase from Comamonas testosteroni was cloned and expressed in an Escherichia coli strain CICC 11022 S with the capability for L-aspartic acid production.The recombinant strain was used for the synthesis of L-alanine. The recombinant strain produced 112. 7g / L L-alanine from fumaric acid with a rate of 12. 5 g /( L·h) and a conversion ratio of 93. 8%. Fumaric acid has a low price,which efficiently reduces the substrate cost for L-alanine production. L-alanine was producd from fumaric acid by using the recombinant strain,and the results showed that the technology had industrial application potential.
引文
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[2]凌关庭.阿力甜及其FAO/WHO(1995)标准[J].中国食品添加剂,1996(2):21-24.
[3]杨海燕,张日生.阿力甜:性质优良的甜味剂[J].中国食品添加剂,2000(3):23-25.
[4]陈天豪,李仁宝,杨威.维生素B6的合成工艺改进[J].中国医药工业杂志,2004,35(1):1-2.
[5]苗华,孙兰英,郭惠元.(S)-2(+)-2-氨基丙醇的制备[J].中国医药工业杂志,1998,29(10):470.
[6]董学伟,张春枝,金凤燮.中空纤维膜反应器转化L-天冬氨酸生成L-丙氨酸[J].食品与发酵工业,2003,29(1):32-35.
[7]ZHAO T,YU B F,TANG W J,et al.Spectrofluorimetric determination of ofloxacin in milk with N-(9-fluorenylmethyloxycarbonyl)-l-alanine[J].Spectrochim Acta A Mol Biomol Spectrosc,2015,148:125-130.
[8]RECH A B,BARIBI G L,VENTURA L H,et al.In vivo dose evaluation during gynaecological radiotherapy using l-alanine/ESR dosimetry[J].Radiat Prot Dosimetry,2014,159(1-4):194-198.
[9]徐虹,王雪根,朱建良,等.L-丙氨酸酶法工业生产的研究[J].工业微生物,1997,27(2):17-20.
[10]TAKAMATSU S,UMEMURA I,YAMAMOTO K,et al.Production of l-alanine from ammonium fumarate using two immobilized microorganisms[J].Eur J Appl Microbiol Biotechnol,1982,15:147-152.
[11]JANDEL A S,HUSTEDT H,WANDREY C.Continuous production of l-alanine from fumarate in a two-stage membrane reactor[J].Eur J Appl Microbiol Biotechnol,1982,15:59-63.
[12]XU Y,CHU H P,GAO C,et al.Systematic metabolic engineering of Escherichia coli for high-yield production of fuel bio-chemical2,3-butanediol[J].Metab Eng,2014,23:22-33.
[13]GRANSSON M,SONDéN B,NILSSON P,et al.Transcriptional silencing and thermoregulation of gene expression in Escherichia coli[J].Nature,1990,344:682-685.