Title of article :
Construction and co-expression of polycistronic plasmid encoding d-hydantoinase and d-carbamoylase for the production of d-amino acids
Author/Authors :
Yangqiu Liu، نويسنده , , Qiang Li، نويسنده , , Xiaojia Hu، نويسنده , , Jichu Yang، نويسنده ,
Issue Information :
روزنامه با شماره پیاپی سال 2008
Abstract :
d-Hydantoinase and d-carbamoylase genes from Agrobacterium radiobacter TH572 were cloned by polymerase chain reaction (PCR). The plasmid pUCCH3 with a polycistronic structure that is controlled by the native hydantoinase promoter was constructed to co-express the two genes and transformed into Escherichia coli strain JM105. To obtain the highest level of expression of the d-carbamoylase and avoid intermediate accumulation, the d-carbamoylase gene was cloned closer to the promoter and the RBS region in the upstream of it was optimized. This resulted in high active expression of soluble d-hydantoinase and d-carbamoylase that is obtained without any inducer. Thus, by the constitutive recombinant JM105/pUCCH3, d-p-hydroxyphenylglycine (d-HPG) was obtained directly with 95.2% production yield and 96.3% conversion yield.
Keywords :
Polycistronic , Co-expression , d-p-Hydroxyphenylglycine , d-Carbamoylase , d-Hydantoinase
Journal title :
Enzyme and Microbial Technology
Journal title :
Enzyme and Microbial Technology