Real-time monitoring of enzymatic hydrolysis of galactomannans

Autor: Joana L. M. S. Ganter, Juan C. Sabbi, Wayne F. Reed
Rok vydání: 2001
Předmět:
Zdroj: Biopolymers. 59:226-242
ISSN: 1097-0282
0006-3525
DOI: 10.1002/1097-0282(20011005)59:4<226::aid-bip1020>3.0.co;2-n
Popis: Enzymatic hydrolysis was monitored in real-time using time dependent static light scattering (TDSLS) for a variety of galactomannans from native Brazilian flora. α-Galactosidase, which strips only the (1–6)α-D galactose side groups, and β-mannanase, which hydrolyses only the (1–4)β-D mannan main chain into oligosaccharides were investigated separately and in combination. The time-dependent signatures matched those describing side-chain stripping for galactosidase, whereas those resulting from the action of mannanase followed the signature typical of random backbone cleavage. Use of both enzymes together required that the TDSLS theory of polymer degradation be extended to the case where random backbone cleavage sites appear as side chains are stripped by the first enzyme. Whereas galactosidase allowed mannanase to access more backbone cleavage sites as time passes, leading to a higher degree of hydrolysis, there was no increase in rate constants. The distribution of random fragments in the case of mannanase digestion alone followed reasonably well the predictions for random cleavage of a single-strand polymer with a restricted number of cleavage sites. The fragment distributions were evaluated by size exclusion chromatography. © 2001 John Wiley & Sons, Inc. Biopolymers 59: 226–242, 2001
Databáze: OpenAIRE
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