Codon Optimization Significantly Improves the Expression Level of α-Amylase Gene from Bacillus licheniformis in Pichia pastoris.

α-Amylase as an important industrial enzyme has been widely used in starch processing, detergent, and paper industries. To improve expression efficiency of recombinant α-amylase from Bacillus licheniformis (B. licheniformis), the α-amylase gene from B. licheniformis was optimized according to the co...

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Publicado en:BioMed Research International Vol. 2015; pp. 1 - 10
Autores principales: Wang, Jian-Rong, Li, Yang-Yuan, Liu, Dan-Ni, Liu, Jing-Shan, Li, Peng, Chen, Li-Zhi, Xu, Shu-De
Formato: pictorial research tables/charts Journal Article
Publicado: Wiley-Blackwell 6/10/2015
Acceso en línea:Ver este registro en EBSCOhost
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      dt: 6/10/2015
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      pub: Wiley-Blackwell
      place: Malden, Massachusetts
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        10.1155/2015/248680
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        atl: Codon Optimization Significantly Improves the Expression Level of α-Amylase Gene from Bacillus licheniformis in Pichia pastoris.
      aug:
        au:
          Wang, Jian-Rong
          Li, Yang-Yuan
          Liu, Dan-Ni
          Liu, Jing-Shan
          Li, Peng
          Chen, Li-Zhi
          Xu, Shu-De
        affil: Guangdong VTR Bio-Tech Co., Ltd., Science and Technology Industry Zone, Nanping, Zhuhai, Guangdong 519060, China
      sug:
        subj:
          Amylases Physiology
          Gene Expression
          Genes
          Biotechnology
          Human
          Funding Source
          Bacillus
          RNA
          Gene Expression Profiling
          Sequence Analysis
          Cell Culture Techniques
      ab: α-Amylase as an important industrial enzyme has been widely used in starch processing, detergent, and paper industries. To improve expression efficiency of recombinant α-amylase from Bacillus licheniformis (B. licheniformis), the α-amylase gene from B. licheniformis was optimized according to the codon usage of Pichia pastoris (P. pastoris) and expressed in P. pastoris. Totally, the codons encoding 305 amino acids were optimized in which a total of 328 nucleotides were changed and the G+C content was increased from 47.6 to 49.2%. The recombinants were cultured in 96-deep-well microplates and screened by a new plate assay method. Compared with the wild-type gene, the optimized gene is expressed at a significantly higher level in P. pastoris after methanol induction for 168 h in 5- and 50-L bioreactor with the maximum activity of 8100 and 11000 U/mL, which was 2.31- and 2.62-fold higher than that by wild-type gene. The improved expression level makes the enzyme a good candidate for α-amylase production in industrial use.
      pubtype: Academic Journal
      doctype:
        pictorial
        research
        tables/charts
        Journal Article
      ougenre: Article
    language: English
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