Molecular Cloning of the Penicillin G Acylase Gene from Arthrobacter Viscosus

Overview

Journal Appl Environ Microbiol

Specialties Environmental Health
Microbiology

Date 1988 Nov 1

PMID 3214149

Citations 10

Authors

H Ohashi

Y Katsuta

T Hashizume

S N Abe

H Kajiura

H Hattori

T Kamei

M Yano

Affiliations

Soon will be listed here.

Abstract

Penicillin G acylase was purified from the cultured filtrate of Arthrobacter viscosus 8895GU and was found to consist of two distinct subunits with apparent molecular weights of 24,000 (alpha) and 60,000 (beta). The partial N-terminal amino acid sequences of the alpha and beta subunits were determined with a protein gas phase sequencer, and a 29-base oligonucleotide corresponding to the partial amino acid sequence of the alpha subunit was synthesized. An Escherichia coli transformant having the penicillin G acylase gene was isolated from an A. viscosus gene library by hybridization with the 29-base probe. The resulting positive clone was further screened by the Serratia marcescens overlay technique. E. coli carrying a plasmid designated pHYM-1 was found to produce penicillin G acylase in the cells. This plasmid had an 8.0-kilobase pair DNA fragment inserted in the EcoRI site of pACYC184.

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