Intracellular co-expression of Vitreoscilla hemoglobin enhances cell performance and ¦Â-galactosidase production in Pichia pastoris

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• 作者：Jyh-Ming Wu ; ^{wzm4@faculty.pccu.edu.tw} ; Wei-Chang Fu ^{FU_95244611@yahoo.com.tw}
• 关键词：Pichia pastoris ; Vitreoscilla hemoglobin ; ¦Â-galactosidase ; AOX1 promoter ; Cell performance ; Recombinant protein expression
• 刊名：Journal of Bioscience and Bioengineering
• 出版年：2012
• 出版时间：March, 2012
• 年：2012
• 卷：113
• 期：3
• 页码：332-337
• 全文大小：503 K

文摘

Pichia pastoris has been used to produce various recombinant proteins under high oxygen demand conditions. To improve the heterologous production of ¦Â-galactosidase, the vgb gene encoding Vitreoscilla hemoglobin (VHb) was co-expressed in the P. pastoris cytoplasm under the control of the methanol-inducible promoter. Co-expression of VHb under different aeration conditions improved cell performance in terms of growth, viability, respiratory rate, and ¦Â-galactosidase production. Under limiting aeration conditions, the VHb⁺ strain produced 28.2 % more biomass but 31.2 % less total ¦Â-galactosidase activity than the VHb^{?/sup> strain. Under non-limiting aeration conditions, the VHb+ strain showed 20.3 % higher cell growth and 9.9 % more total ¦Â-galactosidase activity than the VHb?/sup> strain. Moreover, under these conditions, the VHb+ strain was 7.7 % more viable and had a 28.2 % higher oxygen uptake rate (OUR) than the VHb?/sup> strain. Evidently, VHb can enhance the OUR and promote methanol metabolism, thereby improving cell performance and ¦Â-galactosidase production.}