Production of chorismate mutase-prephenate dehydrogenase by a strain of Escherichia coli carrying a multicopy, tyrA plasmid Isolation and properties of the enzyme》.pdfVIP

6 Biochimica et Biophysica Acta, 717 (1982) 6-11 Elsevier Biomedical Press BBA 21153 PRODUCTION OF CHORISMATE MUTASE-PREPHENATE DEHYDROGENASE BY A STRAIN OF ESCHERICHIA COLI CARRYING A MULTICOPY, tyrA PLASMID ISOLATION AND PROPERTIES OF THE ENZYME SURESH B. BHOSALE *, JUL IAN I. R O O D **, M A R G A R E T K. S N E D D O N and JOHN F. MORRISON *** Department of Biochemistry, John Curtin School of Medical Research, Aust)alian National University, P.O. Box 334, Canberra, A. C T. 2601 (Australia) (Received November 30th, 1981) Key words: Chorismate mutase; Prephenate dehydrogenase; Plasmia~ (E. coil) A multicopy plasmid that contains the tyrosine operon has been used to transform strains of Escherlchia coli K-12. The resultant strains yielded levels-of chorismate mutase-prephenate dehydrogenase that were up to 5000-fold higher than that given by the parent strain and about 6-fold higher than that given by a tyrR strain. The production of enzyme fell when tetracycline was omitted from the growth medium because of the loss of the plasmid. The bifuncfional enzyme was isolated in good yield by a simple purification procedure and shown to possess properties identical to those exhibited by the enzyme from a tyrR strain. Introduction The development of recombinant DNA technol- ogy has opened the way for investigations of the A major problem that has confronted enzymol-