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. 1972 May;110(2):691–698. doi: 10.1128/jb.110.2.691-698.1972

Induction and Characterization of β-Galactosidase in an Extreme Thermophile

J T Ulrich a,1, G A McFeters a, K L Temple a
PMCID: PMC247466  PMID: 5022173

Abstract

A thermostable β-galactosidase (EC 3.2.1.23; β-dgalactoside galactohydrolase) was found to be inducible in an extreme thermophile resembling Thermus aquaticus. Enzyme induction was achieved by the addition of lactose, galactose, or the α-galactoside, melibiose, to growing cultures. The addition of glucose to induced cultures had a repressive effect on further enzyme synthesis. The enzyme was purified 78-fold, and the optimum temperature and pH for activity were determined to be 80 C and pH 5.0, respectively. The enzyme was activated by both manganese and ferrous iron. Sulfhydryl activation and thermal stabilization indicate that the thermophilic β-galactosidase is a sulfhydryl enzyme. Kinetic determinations at 80 C established a Km of 2.0 × 10−3m for the chromogenic substrate o-nitrophenyl β-d-galactopyranoside (ONPG) and a K1 of 7.5 × 10−3m for lactose. The Arrhenius energy of activation (for the hydrolysis of ONPG) was calculated to be 13.7 kcal/mole. A molecular weight of 5.7 × 105 daltons was estimated by elution of the enzyme from Sephadex 4B.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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