Enzyme kinetics of hevamine, a chitinase from the rubber tree Hevea brasiliensis
- 24 July 2000
- journal article
- Published by Wiley in FEBS Letters
- Vol. 478 (1-2) , 119-122
- https://doi.org/10.1016/s0014-5793(00)01833-0
Abstract
The enzyme kinetics of hevamine, a chitinase from the rubber tree Hevea brasiliensis, were studied in detail with a new enzyme assay. In this assay, the enzyme reaction products were derivatized by reductive coupling to a chromophore. Products were separated by HPLC and the amount of product was calculated by peak integration. Penta-N-acetylglucosamine (penta-nag) and hexa-N-acetylglucosamine (hexa-nag) were used as substrates. Hexa-nag was more efficiently converted than penta-nag, which is an indication that hevamine has at least six sugar binding sites in the active site. Tetra-N-acetylglucosamine (tetra-nag) and allosamidin were tested as inhibitors. Allosamidin was found to be a competitive inhibitor with a K i of 3.1 μM. Under the conditions tested, tetra-nag did not inhibit hevamine.Keywords
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