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Overproduction of thermostable leucine dehydrogenase of Bacillus stearothermophilus and its one‐step purification from recombinant cells of Escherichia coli
Author(s) -
Oka M.,
Yang YS,
Nagata S.,
Esaki N.,
Tanaka H.,
Soda K.
Publication year - 1989
Publication title -
biotechnology and applied biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.468
H-Index - 70
eISSN - 1470-8744
pISSN - 0885-4513
DOI - 10.1111/j.1470-8744.1989.tb00063.x
Subject(s) - escherichia coli , thermophile , bacillaceae , recombinant dna , bacillales , biochemistry , enzyme , biology , leucine , plasmid , dehydrogenase , bacillus (shape) , microbiology and biotechnology , bacteria , gene , amino acid , bacillus subtilis , genetics
We have cloned the leucine dehydrogenase (EC 1.4.1.9) gene from a thermophile, Bacillus stearothermophilus, into Escherichia coli MV1184 with a vector plasmid, pUC119. The cloned cells produced a large amount of the thermostable enzyme, which corresponds to about 60% of the total soluble protein. The enzyme was purified to more than 95% homogeneity by only one step, heat treatment of the cell‐extracts, with an average yield of 75 mg/g of wet cells (obtained from 100 ml of the culture).