Mammalian expression levels of cellulase and xylanase genes optimised by human codon usage are not necessarily higher than those optimised by the extremely biased approach

Biotechnol Lett. 2014 Nov;36(11):2169-76. doi: 10.1007/s10529-014-1592-4. Epub 2014 Jun 26.

Abstract

Xylanase gene xynB, cellulase genes egxA and bgl4 were subjected to codon optimisation using two opposing strategies. One was designated the 'one amino acid-one codon' approach, which employs only the codon most used by humans for each amino acid. The other one is referred to as the "humanised" codon usage method, which selects synonymous codons for each amino acid according to the human codon usage table to mimic patterns used in humans. Protein expression levels in mammalian cell lines from each sequence were measured using fluorescence-activated cell sorting, western blotting and enzymatic activity assay. The results indicate that compared with the humanised codon usage method, the relatively simple 'one amino acid-one codon' approach could enhance heterologous protein expression in mammalian cells without apparent drawbacks.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Ascomycota / genetics
  • CHO Cells
  • Cellulase / analysis
  • Cellulase / genetics*
  • Cellulase / metabolism
  • Codon / genetics*
  • Cricetinae
  • Cricetulus
  • Endo-1,4-beta Xylanases / analysis
  • Endo-1,4-beta Xylanases / genetics*
  • Endo-1,4-beta Xylanases / metabolism
  • HEK293 Cells
  • Humans
  • Mollusca / genetics
  • Recombinant Proteins / analysis
  • Recombinant Proteins / genetics*
  • Recombinant Proteins / metabolism
  • Streptomyces / genetics
  • beta-Glucosidase / analysis
  • beta-Glucosidase / genetics*
  • beta-Glucosidase / metabolism

Substances

  • Codon
  • Recombinant Proteins
  • beta-Glucosidase
  • Cellulase
  • Endo-1,4-beta Xylanases
  • XynB xylanase