Improved Production of Aspergillus usamii endo-β-1,4-Xylanase in Pichia pastoris via Combined Strategies.

A series of strategies were applied to improve expression level of recombinant endo-β-1,4-xylanase from Aspergillus usamii (A. usamii) in Pichia pastoris (P. pastoris). Firstly, the endo-β-1,4-xylanase (xynB) gene from A. usamii was optimized for P. pastoris and expressed in P. pastoris. The maximum...

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Publicado en:BioMed Research International Vol. 2016; pp. 1 - 10
Autores principales: Wang, Jianrong, Li, Yangyuan, Liu, Danni
Formato: research tables/charts Journal Article
Publicado: Wiley-Blackwell 3/15/2016
Acceso en línea:Ver este registro en EBSCOhost
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      dt: 3/15/2016
      vid: 2016
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      pub: Wiley-Blackwell
      place: Malden, Massachusetts
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        113767709
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        10.1155/2016/3265895
        113767709
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        atl: Improved Production of Aspergillus usamii endo-β-1,4-Xylanase in Pichia pastoris via Combined Strategies.
      aug:
        au:
          Wang, Jianrong
          Li, Yangyuan
          Liu, Danni
        affil: Guangdong VTR Bio-Tech Co., Ltd., Zhuhai, Guangdong 519060, China
      sug:
        subj:
          Aspergillus
          Yeasts
          Glycoside Hydrolases
          Recombinant Proteins
          Biotechnology Methods
          Descriptive Statistics
          Genes
          Cell Physiology
          Temperature
          Equipment and Supplies
          Genetic Techniques
          Cell Culture Techniques
          Fermentation
          Sequence Analysis
          Oxygen
          Funding Source
      ab: A series of strategies were applied to improve expression level of recombinant endo-β-1,4-xylanase from Aspergillus usamii (A. usamii) in Pichia pastoris (P. pastoris). Firstly, the endo-β-1,4-xylanase (xynB) gene from A. usamii was optimized for P. pastoris and expressed in P. pastoris. The maximum xylanase activity of optimized (xynB-opt) gene was 33500 U/mL after methanol induction for 144 h in 50 L bioreactor, which was 59% higher than that by wild-type (xynB) gene. To further increase the expression of xynB-opt, the Vitreoscilla hemoglobin (VHb) gene was transformed to the recombinant strain containing xynB-opt. The results showed that recombinant strain harboring the xynB-opt and VHb (named X33/xynB-opt-VHb) displayed higher biomass, cell viability, and xylanase activity. The maximum xylanase activity of X33/xynB-opt-VHb in 50 L bioreactor was 45225 U/mL, which was 35% and 115% higher than that by optimized (xynB-opt) gene and wild-type (xynB) gene. Finally, the induction temperature of X33/xynB-opt-VHb was optimized in 50 L bioreactor. The maximum xylanase activity of X33/xynB-opt-VHb reached 58792 U/mL when the induction temperature was 22°C. The results presented here will greatly contribute to improving the production of recombinant proteins in P. pastoris.
      pubtype: Academic Journal
      doctype:
        research
        tables/charts
        Journal Article
      ougenre: Article
    language: English
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