Preparation of the methyl ester of hyaluronan and its enzymatic degradation.
A methyl ester of hyaluronan in which the carboxyl groups were fully esterified was prepared using trimethylsilyl diazomethane. This derivative, while not depolymerized by hyaluronan lyases or hyaluronan hydrolases, was a substrate for both chondroitin ACI lyase (EC 4.2.2.5) from Flavobacterium heparinum and chondroitin ACII lyase (EC 4.2.2.5) from Arthrobacter aurescens. The major product isolated in these depolymerization reactions was methyl alpha-L-threo-hex-4-enepyranosyluronate-(1-->3)-2-acetamido-2-deoxy-alpha,beta-D-glucopyranoside as determined by 1H NMR spectroscopy and MALDITOF mass spectrometry.
Hirano
Kana
K
Graduate School of Pharmaceutical Sciences, Chiba University, 1-33, Inage-ku, Chiba-shi, Chiba 263-8522, Japan.
Sakai
Shinobu
S
Ishikawa
Tsutomu
T
Avci
Fikri Y
FY
Linhardt
Robert J
RJ
Toida
Toshihiko
T
D000818:Animals; D002417:Cattle; D002808:Chondroitin Lyases / Q000378:metabolism; D006820:Hyaluronic Acid / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000378:metabolism; D006821:Hyaluronoglucosaminidase / Q000378:metabolism; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy; D008297:Male; D018502:Streptococcus equi; D013302:Streptomyces / Q000201:enzymology
C036036:hyaluronic acid methyl ester; D006820:Hyaluronic Acid; D006821:Hyaluronoglucosaminidase; D002808:Chondroitin Lyases