Characterization of the catalytic domains of Trichoderma reesei endoglucanase I, II, and III, expressed in Escherichia coli

Hikaru Nakazawa, Katsunori Okada, Ryota Kobayashi, Tetsuya Kubota, Tomoko Onodera, Nobuhiro Ochiai, Naoki Omata, Wataru Ogasawara, Hirofumi Okada, Yasushi Morikawa

Research output: Contribution to journalArticlepeer-review

67 Citations (Scopus)

Abstract

The genes encoding the catalytic domains (CD) of the three endoglucanases (EG I; Cel7B, EG II; Cel5A, and EG III; Cel12A) from Trichoderma reesei QM9414 were expressed in Escherichia coli strains Rosetta-gami B (DE3) pLacI or Origami B (DE3) pLacI and were found to produce functional intracellular proteins. Protein production by the three endoglucanase transformants was evaluated as a function of growth temperature. Maximal productivity of EG I-CD at 15°C, EG II-CD at 20°C and EG III at 37°C resulted in yields of 6.9, 72, and 50 mg/l, respectively. The endoglucanases were purified using a simple purification method based on removing E. coli proteins by isoelectric point precipitation. Specific activity toward carboxymethyl cellulose was found to be 65, 49, and 15 U/mg for EG I-CD, EG II-CD, and EG III, respectively. EG II-CD was able to cleave 1,3-1,4-β-d-glucan and soluble cellulose derivatives. EG III was found to be active against cellulose, 1,3-1,4-β-d-glucan and xyloglucan, while EG I-CD was active against cellulose, 1,3-1,4-β-d-glucan, xyloglucan, xylan, and mannan.

Original languageEnglish
Pages (from-to)681-689
Number of pages9
JournalApplied Microbiology and Biotechnology
Volume81
Issue number4
DOIs
Publication statusPublished - 2008 Dec

Keywords

  • Endoglucanase
  • Heterologous expression
  • Low temperature
  • Rosetta-gami B (DE3) pLacI
  • Trichoderma reesei

ASJC Scopus subject areas

  • Biotechnology
  • Applied Microbiology and Biotechnology

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