Secretory expression and purification of a soluble NADH cytochrome b5 reductase enzyme from Mucorracemosus in Pichiapastoris based on codon usage adaptation

The genome of Mucor racemosus was analyzed to determine the relative levels of codon usage. The codon bias differred from that of Escherichia coli . The active, soluble isoform of NADH cytochrome b5 reductase containing 228 amino acids was successfully overexpressed and secreted using alpha factor i...

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Bibliographic Details
Published in:Biotechnology letters 2010, Vol.32 (11), p.1705-1711
Main Authors: Mirzaei, S. A., Yazdi, M. Tabatabaei, Sepehrizadeh, Z.
Format: Article
Language:English
Subjects:
Applied Microbiology
Biochemistry
Biomedical and Life Sciences
Biotechnology
Life Sciences
Microbiology
Original Research Paper
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Summary:The genome of Mucor racemosus was analyzed to determine the relative levels of codon usage. The codon bias differred from that of Escherichia coli . The active, soluble isoform of NADH cytochrome b5 reductase containing 228 amino acids was successfully overexpressed and secreted using alpha factor in Pichia pastoris under the control of the alcohol oxidase promoter and finally purified. The culture medium and incubation time were optimized, and the maximum expression level observed was about 23 U/ml using X-33 recombinant yeast grown for 120 h with 0.5% (v/v) methanol in complex media.
ISSN:0141-5492
1573-6776
DOI:10.1007/s10529-010-0348-z