High expression of a synthetic gene encoding potato alpha-glucan phosphorylase in Aspergillus niger.
We describe the successful heterologous expression of the Solanum tuberosum alpha-glucan phosphorylase (GP) gene in Aspergillus niger. Special attention was paid to the influence of different codon usage and A+T content in the coding region on GP protein expression. Use of A. niger-preferred codon usage and lower A+T content in a synthetic gene (GP-syn) resulted in a significant improvement in the level of the GP mRNA and a dramatic increase in the quantity of GP protein produced such that it accounted for approximately 10% of the total soluble protein. We suggest that redesigning the primary DNA sequence encoding a desired protein product can be an extremely effective method for improving heterologous protein production in filamentous fungi.
Koda
Akio
A
General Research Laboratory, Ozeki Co., Ltd., 4-9 Imazu, Nishinomiya-shi, Hyogo 663-8227, Japan. akio.kouda@ozeki.co.jp
Bogaki
Takayuki
T
Minetoki
Toshitaka
T
Hirotsune
Masato
M
D001234:Aspergillus niger; D001482:Base Composition; D001483:Base Sequence; D003062:Codon / Q000235:genetics; D015870:Gene Expression; D017343:Genes, Plant / Q000235:genetics; D008969:Molecular Sequence Data; D006005:Phosphorylases / Q000096:biosynthesis* / Q000235:genetics; D010940:Plant Proteins / Q000096:biosynthesis* / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics; D011198:Solanum tuberosum / Q000201:enzymology* / Q000235:genetics
D003062:Codon; D010940:Plant Proteins; D011994:Recombinant Proteins; D006005:Phosphorylases