Conversion of Trp 62 of Hen Egg-White Lysozyme to Tyr by Site-Directed Mutagenesis

An expression plasmid for hen egg-white lysozyme in Saccharomyces cercvisiae was constructed by inserting almost full-length cDNA (about 600 base pairs) encoding hen egg-white pre-lysozyme into a yeast expression vector, pAM 82. The hen lysozyme was expressed under the control of the repressible aci...

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Bibliographic Details
Published in:Journal of biochemistry (Tokyo) 1987-10, Vol.102 (4), p.733-740
Main Authors: KUMAGAI, Izumi, KOJIMA, Shuichi, TAMAKI, Eisuke, MIURA, Kin-ichiro
Format: Article
Language:English
Subjects:
Animals
Base Sequence
Biological and medical sciences
Chickens
Classical genetics, quantitative genetics, hybrids
Cloning, Molecular
Fundamental and applied biological sciences. Psychology
Genetics of eukaryotes. Biological and molecular evolution
Molecular Sequence Data
Muramidase - genetics
Mutation
Saccharomyces cerevisiae - genetics
Thallophyta, bryophyta
Tryptophan
Tyrosine
Vegetals
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Summary:An expression plasmid for hen egg-white lysozyme in Saccharomyces cercvisiae was constructed by inserting almost full-length cDNA (about 600 base pairs) encoding hen egg-white pre-lysozyme into a yeast expression vector, pAM 82. The hen lysozyme was expressed under the control of the repressible acid phosphatase promoter of pAM 82 in S. cerevisiae. About half of the expressed lysozyme was secreted in the yeast growth medium as a precise mature protein which exhibited specific activity consistent with that of authentic hen egg-white lysozyme. The replacement of Trp 62 of hen egg-white lysozyme with a tyrosine residue was performed by site-directed mutagenesis using a 19-mer oligodeoxyribonucleotide. The mutant lysozyme with Tyr 62 was found to exhibit enhanced bacteriolytic activity.
ISSN:0021-924X
1756-2651
DOI:10.1093/oxfordjournals.jbchem.a122111