title: Expression and production of recombinant cis-epoxysuccinate hydrolase in Escherichia coli under the control of temperature-dependent promoter.
abstract: cis-Epoxysuccinate hydrolase (ESH) from Nocardia tartaricans CAS-52 could catalyze the stereospecific hydrolysis of cis-epoxysuccinate to L-(+)-tartrate. The ESH gene of 762 bp was cloned and its open reading frame (ORF) sequence predicted a protein of 253 amino acids. An expression plasmid carrying the ESH gene under the control of the P(L)P(R) promoter was introduced into Escherichia coli, and the ESH gene was successfully expressed in the recombinant strain. The expression conditions and scale-up production were also studied. Fed-batch fermentation of E. coli Trans 1-T1 transformant was carried out in a 2000 L fermentor to product recombinant ESH. The results showed that wet cell concentration reached to 62.45 g L(-1), and the specific activity of ESH was 380.17 U mg(-1), which could meet the requirements of industrial production of L-(+)-tartaric acid.
authors.affiliation: National Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, China.
authors.forename: Ziqiang
authors.initials: Z
authors.lastname: Wang
authors.forename: Yunshan
authors.initials: Y
authors.lastname: Wang
authors.forename: Hong
authors.initials: H
authors.lastname: Shi
authors.forename: Zhiguo
authors.initials: Z
authors.lastname: Su
mesh_terms: D000704:Analysis of Variance; D061249:Batch Cell Culture Techniques; D019149:Bioreactors; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism*; D005285:Fermentation; D005947:Glucose / Q000378:metabolism; D005990:Glycerol / Q000378:metabolism; D006867:Hydrolases / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D013644:Tartrates / Q000378:metabolism; D013696:Temperature
chemical_list: D011994:Recombinant Proteins; D013644:Tartrates; D006867:Hydrolases; C492296:cis-epoxysuccinate hydrolase, Nocardia tartaricans; D005947:Glucose; D005990:Glycerol; C029768:tartaric acid