title: Overproduction of thermostable leucine dehydrogenase of Bacillus stearothermophilus and its one-step purification from recombinant cells of Escherichia coli.
abstract: 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).
authors.affiliation: Institute for Chemical Research, Kyoto University, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Oka
authors.forename: Y S
authors.initials: YS
authors.lastname: Yang
authors.forename: S
authors.initials: S
authors.lastname: Nagata
authors.forename: N
authors.initials: N
authors.lastname: Esaki
authors.forename: H
authors.initials: H
authors.lastname: Tanaka
authors.forename: K
authors.initials: K
authors.lastname: Soda
mesh_terms: D000594:Amino Acid Oxidoreductases / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D002852:Chromatography, Ion Exchange; D003001:Cloning, Molecular; D003720:Densitometry; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D005798:Genes, Bacterial; D005822:Genetic Vectors; D001411:Geobacillus stearothermophilus / Q000201:enzymology*; D006358:Hot Temperature; D005779:Immunodiffusion; D050803:Leucine Dehydrogenase; D010957:Plasmids; D011994:Recombinant Proteins / Q000096:biosynthesis*
chemical_list: D011994:Recombinant Proteins; D000594:Amino Acid Oxidoreductases; D050803:Leucine Dehydrogenase