Cloning, expression and characterization of the esterase estUT1 from Ureibacillus thermosphaericus which belongs to a new lipase family XVIII

Research output: Contribution to journalArticlepeer-review

18 Citations (Scopus)

Abstract

A new esterase gene from thermophilic bacteria Ureibacillus thermosphaericus was cloned into the pET32b vector and expressed in Escherichia coli BL21(DE3). Alignment of the estUT1 amino acid sequence revealed the presence of a novel canonical pentapeptide (GVSLG) and 41–47% identity to the closest family of the bacterial lipases XIII. Thus the esterase estUT1 from U. thermosphaericus was assigned as a member of the novel family XVIII. It also showed a strong activity toward short-chain esters (C2–C8), with the highest activity for C2. When p-nitrophenyl butyrate is used as a substrate, the temperature and pH optimum of the enzyme were 70–80 °C and 8.0, respectively. EstUT1 showed high thermostability and 68.9 ± 2.5% residual activity after incubation at 70 °C for 6 h. Homology modeling of the enzyme structure showed the presence of a putative catalytic triad Ser93, Asp192, and His222. The activity of estUT1 was inhibited by PMSF, suggesting that the serine residue is involved in the catalytic activity of the enzyme. The purified enzyme exhibited high stability in organic solvents. EstUT1 retained 85.8 ± 2.4% residual activity in 30% methanol at 50 °C for 6 h. Stability at high temperature and tolerance to organic solvents make estUT1 a promising enzyme for biotechnology application.

Original languageEnglish
Pages (from-to)271-285
Number of pages15
JournalExtremophiles
Volume22
Issue number2
DOIs
Publication statusPublished - 1 Mar 2018

Keywords

  • Bacteria
  • Esterase
  • Solvent tolerance
  • Thermostability
  • Ureibacillus thermosphaericus
  • CLASSIFICATION
  • BACTERIAL RECOMBINANT LIPASE
  • ALKALINE
  • THERMOSTABLE ESTERASE
  • 1ST MEMBER
  • ENZYMES
  • PSEUDOMONAS-FLUORESCENS CARBOXYLESTERASE
  • PURIFICATION
  • MOLECULAR-CLONING
  • GENE CLONING

Fingerprint Dive into the research topics of 'Cloning, expression and characterization of the esterase estUT1 from Ureibacillus thermosphaericus which belongs to a new lipase family XVIII'. Together they form a unique fingerprint.

Cite this