title: Functional expression of amyloidogenic human stefins A and B in Pichia pastoris using codon optimization.
abstract: Complementary DNAs encoding human stefins A (HSA) and B (HSB) were synthesized using Pichia-preferred codons by overlap extension PCR. The full-length genes were ligated downstream of the glyceraldehyde-3-phosphate dehydrogenase promoter in the Pichia expression vector pGAPZαC and successfully expressed in Pichia pastoris strain X-33. Functional recombinant HSA and HSB proteins were purified from culture medium at yields of 121.3 ± 13.5 (n = 3) and 95.4 ± 4.1 (n = 3) mg/L, respectively. Using this expression strategy, we demonstrated that high levels of bioactive recombinant HSA and HSB can be produced by fermentation in P. pastoris.
authors: Food, Nutrition and Health, University of British Columbia, Vancouver, Canada.
authors: Kosuke
authors: K
authors: Nakamura
authors: Yuki
authors: Y
authors: Maeda
authors: Kensuke
authors: K
authors: Morimoto
authors: Shigeru
authors: S
authors: Katayama
authors: Kazunari
authors: K
authors: Kondo
authors: Soichiro
authors: S
authors: Nakamura
mesh_terms: D000595:Amino Acid Sequence; D058227:Amyloidogenic Proteins / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D003062:Codon / Q000235:genetics*; D055312:Cystatin A / Q000235:genetics* / Q000378:metabolism; D055313:Cystatin B / Q000235:genetics* / Q000378:metabolism; D018076:DNA, Complementary / Q000235:genetics*; D005285:Fermentation / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D010843:Pichia / Q000235:genetics* / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D058227:Amyloidogenic Proteins; C524994:CSTB protein, human; D003062:Codon; D055312:Cystatin A; D018076:DNA, Complementary; D011994:Recombinant Proteins; C026407:CSTA protein, human; D055313:Cystatin B; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases