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1,3-β-d-Glucan synthase ofNeurospora crassa:Partial purification and characterization of solubilized enzyme activity

Authors
Journal
Experimental Mycology
0147-5975
Publisher
Elsevier
Publication Date
Volume
13
Issue
2
Identifiers
DOI: 10.1016/0147-5975(89)90018-2
Disciplines
  • Biology

Abstract

Abstract 1,3-β- d-Glucan synthase activity of Neurospora crassa was rendered soluble by treatment of crude protoplast lysates with 0.1% 3-[3-cholamidopropyl)-dimethylammonio]-1-propanesulfonate and 0.5% octylglucoside in 25 m M 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid buffer containing 5 m M EDTA, 1 m M phenylmethylsulfonyl fluoride, 10 m M sodium fluoride, 1 m M dithiothreitol, 200 m M inorganic phosphate, 10 μ M GTP, and 600 m M glycerol. Approximately 50% of enzyme activity was solubilized; soluble enzyme activity was purified 5.6-fold with a net 38% recovery by sucrose gradient density sedimentation. Partially purified enzyme activity had a half-life of 60 ± 10 h at 4°C, a K m,app of 0.75 ± 0.05 m M, and a V max,app of 35 ± 1 enzyme units/mg protein.

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