Isolation of a β-galactosidase-encoding gene from Bacillus licheniformis

Purification and characterization of the recombinant enzyme expressed in Escherichia coli

Lam Son Phan Trân, Lóránd Szabó, László Fülöp, L. Orosz, Tibor Sík, András Holczinger

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

The Bacillus licheniformis β-galactosidase gene, lacBl, was cloned on a 5.8-kb HindIII fragment into pBR322 and expressed by its own promoter in Escherichia coli. Deletion and complementation analysis showed that the enzyme-encoding region was located on a 4.1-kb HindIII-ClaI fragment. The transcription region for the lacBl was identified on this fragment with promoter- and terminator-probe plasmids. The deduced sequence of 149 aa of the N-terminal part of lacBl showed aa sequence homology with β-Gal from B. stearothermophilus, B. circulans, Haloferax alicantei, Clostridium perfringens, Arthrobacter sp.. No significant homology was shared with those found in the lacZ and lacS families. The recombinant β-galactosidase (LacBl) was purified by FPLC. The molecular mass of the enzyme (80 kDa) and its optimal pH (5.7) and temperature (45°C) were determined.

Original languageEnglish
Pages (from-to)39-43
Number of pages5
JournalCurrent Microbiology
Volume37
Issue number1
DOIs
Publication statusPublished - 1998

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Galactosidases
Haloferax
Escherichia coli
Arthrobacter
Clostridium perfringens
Enzymes
Sequence Homology
Genes
Plasmids
Temperature
Bacillus licheniformis

ASJC Scopus subject areas

  • Microbiology

Cite this

Isolation of a β-galactosidase-encoding gene from Bacillus licheniformis : Purification and characterization of the recombinant enzyme expressed in Escherichia coli. / Trân, Lam Son Phan; Szabó, Lóránd; Fülöp, László; Orosz, L.; Sík, Tibor; Holczinger, András.

In: Current Microbiology, Vol. 37, No. 1, 1998, p. 39-43.

Research output: Contribution to journalArticle

Trân, Lam Son Phan ; Szabó, Lóránd ; Fülöp, László ; Orosz, L. ; Sík, Tibor ; Holczinger, András. / Isolation of a β-galactosidase-encoding gene from Bacillus licheniformis : Purification and characterization of the recombinant enzyme expressed in Escherichia coli. In: Current Microbiology. 1998 ; Vol. 37, No. 1. pp. 39-43.
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