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KMID : 0380219940270060509
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Journal of Biochemistry and Molecular Biology
1994 Volume.27 No. 6 p.509 ~ p.513
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Purification and Characterization of Honey Sucrase
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Abstract
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Abstract:
@EN Sucrase (or invertase)(β-D-fructofuranoside fructohydrolase(EC 3.2.1.26) )was purified from honey by AcellTMlus CM cation exchange chromatography and HPLC-SP column chromatography. The enzyme had a molecular weight of 76,000 daltons, as
determined
by 10% SDS-PAGE. The enzyme showed activity toward sucrose and maltose but did not catalyze the hydrolysis of lactose, raffinose, meelzitose, inulin, starch, p-nitrophenyl-a-D-glucopyranoside(a-PNPG), or
p=nitrophenyl-β-D-glucopyranoside(β-PNPG).
The
Vmax and Km values of purified sucrase against sucrose were 100 U per mg. Of protein and 91.2mM, respectively, and against maltose they were 31,25 U per mg, of protein and 60 mM, respectively. The optimum pH and temperature of the enzyme were pH
5.0~6.0and 40~50℃, respectively. When purified honey sucrase was added to a reaction mixture containing maltose or sucrose, a large amount of monosaccharide was produced, but trisaccharide was not detected. Honey sucrase was inhibited by metal
ions and
chemical modifiers, such as Hg2+, I2, 1-fluoro 2,4-dinitrobenzene (FDNB), ρ-hydroxymercuribenzoic acid (HMB), and n=bromosuccinimide(NBS), but not by D-fructose, the sucrose hydrolytic product.
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