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Volume 140, Issue 9

Characterization of chitin synthase from Free

Abstract

Chitin synthase in a microsomal preparation from had an apparent for UDP--acetylglucosamine of 2.0 mM while nikkomycin Z and polyoxin D inhibited enzyme activity competitively with apparent values of approximately 0.1 μM and 6 μM respectively. The organophosphorus fungicide edifenphos was a non-competitive inhibitor ( 54 μM). Preincubation of microsomes for 2 h at 25 °C resulted in a maximum twofold stimulation of chitin synthase activity while preincubation with trypsin (25 μg ml) or cytosol (350 μg cytosolic protein ml) for 10 min at 25 °C resulted in approximately fourfold and 20-fold increases in chitin synthase activity, respectively. A range of protease inhibitors reduced the degree of activation of microsomal chitin synthase by cytosol. Most potent were phenylmethanesulphonyl fluoride and chymostatin; these compounds completely inhibited activation of enzyme activity. Two fragments (approx. 600 bp; and ) were amplified from genomic DNA using degenerate PCR primers based on regions of complete amino acid homology between previously published chitin synthase gene sequences. When the DNA and predicted amino acid sequences of were used to probe computer databases for related sequences, was found to be most similar to from .

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Keyword(s): Botrytis cinerea , chitin synthase , phylogenetic analysis , polymerase chain reaction and zymogen
© Society for General Microbiology 1994
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1994-09-01
2024-05-03
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Characterization of chitin synthase from Botrytis cinerea
Microbiology 140, 2199 (1994); https://doi.org/10.1099/13500872-140-9-2199
/content/journal/micro/10.1099/13500872-140-9-2199
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