PURIFICATION AND PROPERTIES OF ANTHRANILATE SYNTHETASE FROM ERGOT FUNGUS, CLAVICEPS-SPEC, STRAIN SD 58

  • 1 January 1977
    • journal article
    • research article
    • Vol. 40  (2) , 136-145
Abstract
A 3-enzyme complex containing anthranilate synthetase, phosphoribosyl anthranilate isomerase and indole-3-glycerol phosphate synthetase was partially purified from Claviceps sp., strain SD 58. The anthranilate synthetase activity of the enzyme complex was quite unstable unless glutamine, MgCl2 TRIS and glycerol were included in the extraction buffer. The 3-enzyme complex showed a MW of 400,000 when estimated using Sephadex gel filtration and a MW of 200,000 when using sucrose density gradient centrifugation. At least 2 bands of anthranilate synthetase were detected on disc gel electrophoresis. An enzyme complex containing phosphoribosyl anthranilate synthetase and indoleglycerol phosphate synthetase, but no anthranilate synthetase, was isolated from Claviceps. This enzyme complex had an apparent MW of 165,000 as estimated by sucrose gradient centrifugation. Anthranilate synthetase is inhibited by L-tryptophan and elymoclavine, the terminal ergot alkaloid produced by this strain of Claviceps. No differences were detected between the enzyme complexes isolated from 2 day old growing mycelia and from 6 day old alkaloid-producing mycelia of the organism.