Partial marker cassette mutagenesis of xylose isomerase

Details Partial marker cassette mutagenesis of xylose isomerase Partial marker cassette mutagenesis of xylose isomerase

1985-09-27

1990-01-16

Wiseman, Thomas G.

Chemistry: molecular biology and microbiology

Micro-organism, tissue cell culture or enzyme using process...

Preparing compound containing saccharide radical

4351725, 435234, 435320, 43525233, 536 27, 935 14, 935 29, 935 61, C12P 2100, C12P 1924, C12N 1500, C12N 992

Patent

active

048943312

ABSTRACT:
Constructions and methods for mutagenesis of nucleic acids involve chemical mutagenesis of a cassette comprising a structural gene linked to a non-functional restorable fragment of a marker gene. Mutants are detected by screening for the presence of the reconstituted marker among the ligation products of the cassette to a vector containing the non-functional restorable remainder of the marker gene. Xylose isomerase mutants, characterized by a change from glu (GAG) to lys (AAG) at amino acid position 262 in the xylA protein of E. coli were obtained by partial marker cassette mutagenesis. These mutants enzymes exhibited twice the rate of isomerization of glucose to fructose exhibited by the wild type.

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