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Molecular Cloning of a β-Glucosidase Gene from an Extremely Thermophilic Anaerobe in E. coli and B. subtilis

Bio/Technology volume 5, pages 384–387 (1987)Cite this article

Abstract

A genomic library of an extremely thermophilic anaerobic bacterium, Caldocellum saccharolyticum, was constructed using λ1059 as the cloning vector. The chromogenic substrate 5–bromo–4–chloro–3–indolyl–β–D–glucopyranoside (BCIGlucose) was used for the in situ detection of cloned β–glucosidase. Plasmid subcloning showed that the β–glucosidase gene was present in a 1.78 kb HindIII fragment. The expression of this gene in E. coli and B. subtilis using pBR322 and pC194 as vectors was dependent on orientation, suggesting that transcription may be initiated from a vector promoter sequence. β–glucosidase activity expressed in the mesophilic host cells was cell–associated. Assays of β–glucosidase activity, in toluene–treated cells of transformed E. coli, showed a temperature maximum of 85°C, and a pH maximum of 6.25. The electrophoretic mobility of the β–glucosidase in a heat–treated extract of transformed E. coli maxicells indicated an Mr of 52,000.

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  1. Donald R. Love: Corresponding author.

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  1. Genetics Research Unit, Department of Cell Biology, University of Auckland, Private Bag, Auckland, New Zealand

    Donald R. Love & Markus B. Streiff

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Love, D., Streiff, M. Molecular Cloning of a β-Glucosidase Gene from an Extremely Thermophilic Anaerobe in E. coli and B. subtilis. Nat Biotechnol 5, 384–387 (1987). https://doi.org/10.1038/nbt0487-384

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