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Research Article


ScienceAsia 20 (1994): 145-155 |doi: 10.2306/scienceasia1513-1874.1994.20.145

 

PRODUCTION OF SORBITOL AND ETHANOL FROM SUCROSE BY ZYMOMONAS MOBILIS: SEPARATION AND PROPERTIES OF GLUCOSE-FRUCTOSE OXIDOREDUCTASE

 

BOONYRAS SOOKHEO,a SUREE PHUTRAKULb AND PAWINEE KANASAWUDb

ABSTRACT: Sorbitol was found as a product from reduction of fructose by NADH catalyzing with intracellular enzyme extracted from Zymomonas mobilis IFO.13756. Grinding with sand provided the maximum activity of glucose-fructose oxidoreductase comparing to the others four methods of extraction; treatment of lysozyme, cell disruption with ultrasonic wave, extraction with cold acetone and extraction of cell debris by Triton X-100. Partial purification of enzyme by Blue-Sepharose CL-4B column resulted in 7.22 folds of purification with 23.39% yield which was higher than the purification by DEAE Sephadex A-50. The partial purified enzyme from the higher yield method gave the optimum pH, optimum temperature, V max and Km at 7.0, 25oC, 0.006 mole. min-1 and 4 mM respectively. The catalytic reduction property of the partial purified enzyme was confirmed by the activity assay of the protein band on polyacrylamide gel electrophoresis.

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a Faculty of Science, Khrungthep University, Bangkok, Thailand.
b Faculty of Science, Chiang Mai University, Chiang Mai, Thailand.

Received June 23, 1994