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KMID : 1134820110400091321
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Journal of the Korean Society of Food Science and Nutrition
2011 Volume.40 No. 9 p.1321 ~ p.1327
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Purification and Characterization of an Invertase Produced with Saccharomyces cerevisiae JS59 Isolated from Home-made Wine
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Yoo Ji-Soo
Paik Hyun-Dong
Kim Soo-Young
Lee Si-Kyung
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Abstract
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The microorganism producing an invertase (E.C. 3.2.1.26) was isolated from wine and tentatively identified as Saccharomyces cerevisiae by cellular fatty acid analysis. The invertase was purified to homogeneity by ammonium sulfate precipitant, dialysis, ion-exchange chromatography on DEAE-Sephadex A-50, and gel chromatography on Sephadex G-200 from the culture supernatant of Saccharomyces cerevisiae JS59. The specific activity and the purification fold of the purified invertase were 7620.9 unit/mg protein and 13.9, respectively. The molecular weight of the purified invertase was estimated to be 38.5 kDa by SDS-PAGE. The optimum pH and temperature for the invertase activity were pH 5 and 55℃, respectively. The invertase activity was relatively stable at pH 4～6 and temperature 55℃. The activity of invertase was inhibited by Ag2+ and Hg2+, but on the contrary, activated by Co2+ and Mn2+. Michaelis constant (Km) for invertase reaction in sucrose solution was 11.5 mM. TLC analysis of the products produced in sucrose solution during invertase reaction showed the progressive presence of glucose and fructose in accordance with sucrose hydrolysis.
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KEYWORD
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invertase, Sacchromyces cerevisiae, purification, TLC analysis
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