A chitinolytic organism was cultured from a lotic habitat in central Indiana. It was isolated on the basis of its hydrolytic activity on chitin agar. It was subsequently identified as EF-4a. The chitinases of the EF-4a were isolated, purified and concentrated. A colloidal chitin suspension was then incubated with the enzyme. Samplings were taken at one minute, one hour and 24 hours. The samplings were filtered through a Swinney filtration unit and immediately after applied to a Beckman Ultrasil Amino column. It was eluted with acetonitrile: water (75:25) at a flow rate of 2 ml/min. The absorbance at 214 nm was monitored using the Beckman 160 UV detector. The chromatographs were analyzed for product elaboration. It was observed that EF-4a produced soluble, extracellular enzymes. The enzymes were a chitinase-chitobiase complex. The enzymes' action on chitin was immediate. There were varying reaction products including G1cN, GlcNAc, chitobiose, chitotriose and higher order oligomers. There was also chromatographic evidence suggesting a and B anomers of these saccharides.
| Identifer | oai:union.ndltd.org:BSU/oai:cardinalscholar.bsu.edu:handle/183132 |
| Date | January 1985 |
| Creators | Baker-Ellis, Judy |
| Contributors | Warnes, Carl E. |
| Source Sets | Ball State University |
| Detected Language | English |
| Format | v, 54 leaves : ill. ; 28 cm. |
| Source | Virtual Press |
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