Cloning and characterization of β-galactoside and β-glucoside hydrolysing enzymes of Thermotoga maritima

Josef Gabelsberger, Wolfgang Liebl, Karl Heinz Schleifer

Research output: Contribution to journalArticlepeer-review

48 Scopus citations

Abstract

A gene library of the hyperthermophilic bacterium Thermotoga maritima strain MSB8 was constructed in Escherichia coli. Two non-related T. maritima chromosomal DNA fragments were physically characterized. They conferred the synthesis of thermostable X-Gal (5-bromo-4-chloro-3-indolyl-β-d-galactopyranoside)-hydrolysing activity upon the host organism. The biochemical properties of the recombinant enzymes indicated that genes for a β-galactosidase (BgaA) and a broad-specificity β-glucosidase (Bg1A) had been isolated. The genes were desiignted bgaA and bglA, respectively. According to analytical size exclusion chromatography data, BgaA and BglA had native molecular masses of approximately 240 kDa and 95 kDa, respectively. Both enzymes apparently have dimeric subunit structure. An additional β-glucosidase (designated BglB) activity, clearly distinct from BglA in terms of substrate specificity, could be detected in a crude extract of T. maritima.

Original languageEnglish
Pages (from-to)131-137
Number of pages7
JournalFEMS Microbiology Letters
Volume109
Issue number2-3
DOIs
StatePublished - 15 May 1993
Externally publishedYes

Keywords

  • Thermostable enzyme
  • Thermotoga maritima
  • β-Galactosidase
  • β-Glucosidase

Fingerprint

Dive into the research topics of 'Cloning and characterization of β-galactoside and β-glucoside hydrolysing enzymes of Thermotoga maritima'. Together they form a unique fingerprint.

Cite this