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Production of d ‐ p ‐Hydroxyphenylglycine by N ‐Carbamoyl‐ d ‐amino Acid Amidohydrolase‐Overproducing Escherichia coli Strains
Author(s) -
Chao YunPeng,
Juang TzongYuan,
Chern JongTzer,
Lee ChengKang
Publication year - 1999
Publication title -
biotechnology progress
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 129
eISSN - 1520-6033
pISSN - 8756-7938
DOI - 10.1021/bp990060c
Subject(s) - escherichia coli , amidohydrolase , biology , plasmid , recombinant dna , transformation (genetics) , biochemistry , strain (injury) , subcloning , amino acid , enzyme , gene , anatomy
The N ‐carbamoyl‐ d ‐amino acid amidohydrolase ( d ‐carbamoylase) gene from Agrobacterium radiobacter NRRL B11291 has been successfully cloned and expressed in Escherichia coli. Subcloning of the d ‐carbamoylase gene into different types of vectors and backgrounds of E. coli strains showed that the optimal expression level of d ‐carbamoylase was achieved in a ColE1‐derived plasmid with a 150‐fold increase in specific enzyme activity compared to that in a pSC101‐derived plasmid. In addition, the recombinant plasmids were very stable in the E. coli strain ATCC11303 but not in JCL1258 tested here. Employing the recombinant E. coli strain DH5α/pAH61 for d ‐ p ‐hydroxyphenylglycine production showed that the cell was capable of transforming N ‐carbamoyl‐ d ‐hydroxylphenylglycine to d ‐ p ‐hydroxyphenylglycine with a molar conversion yield of 100% and a production rate of 1.9 g/(L h). In comparison with A. radiobacter NRRL B11291, this productivity approximates a 55‐fold increase in d ‐hydroxyphenylglycine production. This result suggests the potential application of recombinant E. coli strains for the transformation reaction.