THERMOSTABLE GLYCEROL KINASE FROM THERMUS-FLAVUS - CLONING, SEQUENCING, AND EXPRESSION OF THE ENZYME GENE

The thermostable glycerol kinase (EC 2.7.1.30) gene from Thermus flavu s was cloned and expressed in Escherichia toll DH5 alpha. An open read ing frame of 1488 bp for the glycerol kinase gene (glpK) starting with an ATG methionine codon was found, which encodes a protein of 496 ami no acid residues whose calculated molecular weight is 54,835. The amin o acid sequence of T. flavus glycerol kinase is 80.6% and 64.1% identi cal with those of Bacillus subtilis and E. coli. Transformants of E. c oli DH5 alpha harboring plasmid pGYK12 with a 1505 bp chromosomal DNA fragment containing the T. flavus glycerol kinase gene showed about 23 .8-fold higher glycerol kinase activity than T. flavus. (C) 1998 Elsev ier Science B.V.