Cloning and expression of Cel8A from Klebsiella pneumoniae in Escherichia coli and comparison to cel gene of Cellulomonas uda

I. Son Ng, Xiaoqin Chi, Xiaomin Wu, Ziwei Bao, Yinghua Lu, Jo Shu Chang, Xueping Ling

Research output: Contribution to journalArticlepeer-review

9 Citations (Scopus)

Abstract

Klebsiella pneumoniae XM-4 is first isolated from a bacterial consortium. According to the 16S rRNA sequence, it has only 75% sequence similarity to Cellulomonas sp. However, a 999-bp open reading frame of K. pneumoniae, which encodes a putative endo-glucanase of 333 amino acids belonging to GH8 family and designated as Cel8A, is found closely to Cellulomonas uda with 95% similarity in amino acid sequence. This implies that the biological evolution between both strains has occurred. The cel8A is constructed in vectors of pET-22b(+) and pET-32a(+) with or without containing a signal peptide, and then cloned and expressed in Escherichia coli. The best endo-glucanase production of 25.4. mg/l is then obtainable by applying pET-32a(+) without the signal peptide. The recombinant CelA8 has an optimal specific activity of 62.4. U/mg against CMC or 23.3. U/mg against β-. d-glucan at 55. °C and pH 5.0, indicating good potential for the industrial application.

Original languageEnglish
Pages (from-to)53-58
Number of pages6
JournalBiochemical Engineering Journal
Volume78
DOIs
Publication statusPublished - 2013 Sep 5

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Environmental Engineering
  • Bioengineering
  • Biomedical Engineering

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