Solvent and viscosity effects on the rate-limiting product release step of glucoamylase during maltose hydrolysis

M. R. Sierks, C. Sico, M. Zaw

Research output: Contribution to journalArticle

23 Scopus citations

Abstract

Release of product from the active site is the rate-limiting step in a number of enzymatic reactions, including maltose hydrolysis by glucoamylase (GA). With GA, an enzymatic conformational change has been associated with the product release step. Solvent characteristics such as viscosity can strongly influence protein conformational changes. Here we show that the rate-limiting step of GA has a rather complex dependence on solvent characteristics. Seven different cosolvents were added to the GA/maltose reaction solution. Five of the cosolvents, all having an ethylene glycol base, resulted in an increase in activity at low concentration of cosolvent and variable decreases in activity at higher concentrations. The increase in enzyme activity was dependent on polymer length of the cosolvent; the longer the polymer, the lower the concentration needed. The maximum increase in catalytic activity at 45 °C (40-45%) was obtained with the three longest polymers (degree of polymerization from 200 to 8000). A further increase inactivity to 60-65% was obtained at 60 °C. The linear relationship between ln(k(cat)) and (viscosity)2 obtained with all the cosolvents provides further evidence that product release is the rate-limiting step in the GA catalytic mechanism. A substantial increase in the turnover rate of GA by addition of relatively small amounts of a cosolvent has potential applications for the food industry where high-fructose corn syrup (HFCS) is one of the primary products produced with GA. Since maltodextrin hydrolysis by GA is by far the slowest step in the production of HFCS, increasing the catalytic rate of GA can substantially reduce the process time.

Original languageEnglish (US)
Pages (from-to)601-608
Number of pages8
JournalBiotechnology Progress
Volume13
Issue number5
DOIs
StatePublished - Sep 1 1997
Externally publishedYes

ASJC Scopus subject areas

  • Biotechnology

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