Premium
A Cultivation Strategy of Recombinant Escherichia coli for Mass Production of Thermostable d‐Hydantoinase
Author(s) -
LEE DONGCHEOL,
KIM GEUNJOONG,
CHA YOOKYONG,
LEE CHEOYOUNG,
KIM HAKSUNG
Publication year - 1998
Publication title -
annals of the new york academy of sciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.712
H-Index - 248
eISSN - 1749-6632
pISSN - 0077-8923
DOI - 10.1111/j.1749-6632.1998.tb10342.x
Subject(s) - library science , corporation , political science , computer science , law
Optically pure D-amino acids are widely used in the pharmaceutical field as intermediates for the synthesis of semisynthetic antibiotics, peptide hormones, pyrethroids, and pesticides. Currently, D-hydantoinase (EC 3.5.2.2) is employed as an industrial biocatalyst for the synthesis of the D-amino acids.1 We have focused on the screening of thermostable D-hydantoinase-producing microorganisms and have isolated an enzyme from Bacillus stearothermophilus SD-1.2 The enzyme has been found to be most thermostable among the D-hydantoinases reported so far.3 The gene encoding the enzyme was previously cloned and constitutively overexpressed in Escherichia coli XL1-Blue/pHU183 by its native promoter in a soluble form.4 Mukohara et al. also reported the expression in E. coli of a thermostable D-hydantoinase gene from B. stearothermophilus NS1122A by an inductive promoter, but in this case the enzyme was produced as an insoluble aggregate.5 In order for an industrial enzyme to be practically applicable, the development of a cost-effective process for mass production of the enzyme should be given the highest priority. In this work, we attempted the mass production of the whole cell enzyme of thermostable D-hydantoinase using the recombinant E. coli. The harvested cells can be directly employed as a biocatalyst in the enzymatic process.