Int. J. Mol. Sci. 2011, 12(5), 3366-3380; doi:10.3390/ijms12053366
Article

Expression of an Endo-β-1,4-glucanase Gene from Orpinomyces PC-2 in Pichia pastoris

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Received: 11 April 2011; in revised form: 3 May 2011 / Accepted: 23 May 2011 / Published: 24 May 2011
(This article belongs to the Section Biochemistry, Molecular Biology and Biophysics)
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract: The endo-β-1,4-glucanase gene celE from the anaerobic fungus Orpinomyces PC-2 was placed under the control of an alcohol oxidase promoter (AOX1) in the plasmid pPIC9K, and integrated into the genome of a methylotrophic yeast P. pastoris GS115 by electroporation. The strain with highest endo-β-1,4-glucanase activity was selected and designed as P. pastoris egE, and cultivated in shaking flasks. The culture supernatant was assayed by SDS-polyacrylamide gel electrophoresis and showed a single band at about 52 kDa. Furthermore, the recombinant P. pastoris egE was proved to possess the ability to utilize sodium carboxymethyl cellulose as a carbon source. The recombinant endoglucanase produced by P. pastoris showed maximum activity at pH 6.0 and temperature 45 °C, indicating it was a mesophilic neutral endo-β-1,4-glucanase, suitable for denim biofinishing/washing. Further research was carried out in suitable fermentation medium in shaking flasks. The most favorable methanol addition concentration was discussed and given as 1.0%. After methanol induction for 96 h, the endo-β-1,4-glucanase activity reached 72.5 IU mL−1. This is the first report on expression and characterization of endo-β-1,4-glucanase from Orpinomyces in P. pastoris. The endo-β-1,4-glucanase secreted by recombinant P. pastoris represents an attractive potential for both academic research and textile industry application.
Keywords: endo-β-1,4-glucanase; Pichia pastoris; heterologous expression; neutral cellulase; induction medium
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MDPI and ACS Style

Jin, X.; Meng, N.; Xia, L.-M. Expression of an Endo-β-1,4-glucanase Gene from Orpinomyces PC-2 in Pichia pastoris. Int. J. Mol. Sci. 2011, 12, 3366-3380.

AMA Style

Jin X, Meng N, Xia L-M. Expression of an Endo-β-1,4-glucanase Gene from Orpinomyces PC-2 in Pichia pastoris. International Journal of Molecular Sciences. 2011; 12(5):3366-3380.

Chicago/Turabian Style

Jin, Xin; Meng, Nan; Xia, Li-ming. 2011. "Expression of an Endo-β-1,4-glucanase Gene from Orpinomyces PC-2 in Pichia pastoris." Int. J. Mol. Sci. 12, no. 5: 3366-3380.

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