Cloning, Expression and Characterization of Endo-beta-1,4-Mannanase from Aspergillus fumigatus in Aspergillus sojae and Pichia pastoris

Date

2009-01-01

Author

Duruksu, Goekhan
Ozturk, Bengu
Biely, Peter
Bakir, Ufuk
Ögel, Zümrüt Begüm

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To be utilized in biomass conversion, including ethanol production and galactosylated oligosaccharide synthesis, namely prebiotics, the gene of extracellular endo-beta-1,4-mannanase (EC 3.2.1.78) of Aspergillus fumigatus IMI 385708 (formerly known as Thermomyces lanuginosus IMI 158749) was expressed first ill Aspergillus sojae and then in Pichia pastoris under the control of the glyceraldehyde triphosphate dehydrogenase ((gpdA) under bar) and the alcohol oxidase ((AOX1) under bar) promoters, respectively. The highest production of mannanase (352 U mL(-l)) in A. sojae was observed after 6 days of cultivation. In P. pastoris, the highest mannanase production was observed 10 h after induction with methanol (61 U mL(-1) The fold increase in mannanase production was estimated as similar to 12-fold and similar to 2-fold in A. sojae and P. pastoris, respectively, when compared with A. fumigatus. Both recombinant enzymes showed molecular mass of about 60 kDa and similar specific activities (similar to 350 U mg(-1) protein). Temperature optima were at 60 degrees C and 45 degrees C, and maximum activity was at pH 4.5 and 5.2 for A. sojae and P. pastoris, respectively. The enzyme from P. pastoris was more stable retaining most of the activity up to 50 degrees C, whereas the enzyme from A. sojae rapidly lost activity above 40 degrees C. (C) 2009 American Institute of Chemical Engineers Biotechnol. Prog., 25: 271-276, 2009

Subject Keywords

Beta-Mannanase, Overproduction, Aspergillus fumigatus, Pichia pastoris, Aspergillus sojae

URI

https://hdl.handle.net/11511/49801

Journal

BIOTECHNOLOGY PROGRESS

DOI

https://doi.org/10.1021/bp.104

Collections

Graduate School of Natural and Applied Sciences, Article

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Citation Formats

G. Duruksu, B. Ozturk, P. Biely, U. Bakir, and Z. B. Ögel, “Cloning, Expression and Characterization of Endo-beta-1,4-Mannanase from Aspergillus fumigatus in Aspergillus sojae and Pichia pastoris,” BIOTECHNOLOGY PROGRESS, pp. 271–276, 2009, Accessed: 00, 2020. [Online]. Available: https://hdl.handle.net/11511/49801.