Effect of Co-expression of Vitreoscilla Hemoglobin on Expression of Β-mannanase in Pichia pastoris

doi:10.13560/j.cnki.biotech.bull.1985.2015.12.028

Abstract

Abstract:

In order to improve the limitation of dissolved oxygen and increase Β-mannanase production in the process of high-density fermentation, the Β-mannanase gene and Vitreoscilla hemoglobin gene(VHb)were co-expressed in Pichia pastoris under the control of AOX1 promoter. The VHb gene was synthesized by codon optimization and inserted into expression vector pPICZαA, then integrated into the engineering bacterium of Β-mannanase gene, P. pastoris GS115/Ppic9K-Β-MAN. The recombinant P. pastoris of co-expressing VHb was selected by G418 and Zeocin resistance screening. Finally, the discrepancies in Β-mannanase expression between the co-expressed strain VHb⁺ and initial strain VHb^-were analyzed in 30 L fermenter. The results showed as below, compared with the controlled strain, the expression of Β-mannanase by VHb⁺ strain was increased by 90% under oxygen-limited condition. Furthermore, Vitreoscilla hemoglobin was improved on the methanol tolerance, and fermentation period was shorted about 40 h, which could be an important industrial value.

Key words: Vitreoscilla hemoglobin, &#x00392, -mannanase, Pichia pastoris, high-density fermentation

Zhang Xiaolong, Xiao Jing, Wang Ruiming, Wang Junqing, Wang Xingji, Wang Zhengxiang. Effect of Co-expression of Vitreoscilla Hemoglobin on Expression of Β-mannanase in Pichia pastoris[J]. Biotechnology Bulletin, 2015, 31(12): 193-199.

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