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Title 

Screening system for chitin synthase II inhibitors from natural resources and its inhibitor prodigiosin

Authors 

Eui Il HwangYoung Kook KimHyang Bok LeeHong Gi KimSung Uk Kim

Publisher 

The Korean Society for Applied Microbiology

Issue Date 

2000

Citation 

Journal of Microbiology and Biotechnology, vol. 10, no. 2, pp. 251-257

Keywords 

chitin synthasechitin synthase II inhibitorantifungal agentsprodigiosin

Abstract 

Chitin synthases are identified as key enzymes of chitin biosynthesis in most of the fungi. Among them, chitin synthase II has been reported to be an essential enzyme in chitin biosynthesis, and exists as a membrane-bound form. To search and screen new antifungal agents from natural resources to inhibit chitin synthase II, the assay conditions were established using the enzyme isolated from Saccharomyces cerevisiae ECY38-38A(pAS6) that overproduces only chitin synthase II. This enzyme was activated only by partial proteolysis with trypsin. Its activity reached the maximum at 80 μg/ml of trypsin and was strongly stimulated by 2.0 mM Co2+, 1.0 mM UDP-[14C]-GlcNAc, and 32 mM free-GlcNAc. Under these assay conditions, the highest chitin synthase II activity was observed by incubation at 30°C for 90 min. However, an extremely narrow range of organic solvents up to as much as 25% of DMSO and 25% of MeOH was useful for determining optimal assay conditions. After a search for potent inhibitors of chitin synthase II from natural resources, prodigiosin was isolated from Serratia marcescens and purified by solvent extraction and silica gel column chromatographies. The structure of prodigiosin was determined by UV, IR, Mass spectral, and NMR spectral analyses. Its molecular weight and formula were found to be 323 and C20H25N3O, respectively. Prodigiosin inhibited chitin synthase II by 50% at the concentration of 115 μg/ml.

ISSN 

1017-7825

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2017-04-19


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