Improved glutathione production by gene expression in Pichia pastoris.
To utilize Pichia pastoris to produce glutathione, an intracellular expression vector harboring two genes (gsh1 and gsh2) from Saccharomyces cerevisiae encoding enzymes involved in glutathione synthesis and regulated by the glyceraldehyde-3-phosphate dehydrogenase (GAP) promoter was transformed into P. pastoris GS115. Through Zeocin resistance and expression screening, a transformant that had higher glutathione yield (217 mg/L) in flask culture than the host strain was obtained. In fed-batch culture process, this recombinant strain displayed high activity for converting precursor amino acids into glutathione. The glutathione yield and biomass achieved 4.15 g/L and 98.15 g (dry cell weight, DCW)/L, respectively, after 50 h fermentation combined with addition of three amino acids (15 mmol/L glutamic acid, 15 mmol/L cysteine, and 15 mmol/L glycine).
Fei
Liwen
L
Department of Biochemistry, Shanghai Institute of Pharmaceutical Industry, 200040 Shanghai, People's Republic of China.
Wang
Yan
Y
Chen
Shaoxin
S
D001483:Base Sequence; D019149:Bioreactors / Q000382:microbiology; D017931:DNA Primers / Q000235:genetics; D015870:Gene Expression; D005818:Genetic Engineering; D005822:Genetic Vectors; D005721:Glutamate-Cysteine Ligase / Q000235:genetics / Q000378:metabolism; D005978:Glutathione / Q000096:biosynthesis*; D010843:Pichia / Q000235:genetics* / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism
D017931:DNA Primers; D011994:Recombinant Proteins; D029701:Saccharomyces cerevisiae Proteins; C490862:GSH1 protein, S cerevisiae; D005721:Glutamate-Cysteine Ligase; D005978:Glutathione