PMID: 2503013Jun 1, 1989Paper

Overproduction of thermostable leucine dehydrogenase of Bacillus stearothermophilus and its one-step purification from recombinant cells of Escherichia coli

Biotechnology and Applied Biochemistry
M OkaK Soda

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).

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