Human alpha atrial natriuretic peptide fermentation by using a genetically engineered yeast strain.
The genotype of the recombinant yeast strain Y33::YFD71-3 used in this study is alpha, his, leu, ade and suc. Preliminary batch cultures in shaking flasks showed the expression level of atrial natriuretic peptide (ANP) was 1-2 mg/L. Shaking flask cultures were carried out in YG medium which contained glucose, yeast nitrogen base (YNB), and different amounts of adenine, histidine and leucine. When the growth was limited by adenine, protein secreted from Y33::YFD71-3 cells was increased obviously. Adenine became the limiting substrate when the YG medium was supplemented with 5 g/L of casamino acid (CAA) and the level of ANP expression was influenced by the concentrations of adenine, YNB and leucine in the medium. In fed batch cultures carried out in a RIBE-5 fermenter, the cultures were fed with glucose and a mixture of YNB, CAA, adenine, histidine and leucine to improve cell growth and ANP expression, and the maximum ANP concentration in the culture reached 24.8 mg/L.
Ye
Q
Q
Research Institute of Biochemical Engineering, East China University of Science and Technology, Shanghai, China.
Tao
J
J
Zhang
H
H
Zhang
S
S
Cai
H
H
Bao
W
W
Li
Y
Y
D000225:Adenine / Q000494:pharmacology; D000596:Amino Acids / Q000494:pharmacology; D009320:Atrial Natriuretic Factor / Q000096:biosynthesis*; D003470:Culture Media; D004305:Dose-Response Relationship, Drug; D005285:Fermentation; D006639:Histidine / Q000494:pharmacology; D006801:Humans; D007930:Leucine / Q000494:pharmacology; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis*; D015003:Yeasts / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism
D000596:Amino Acids; D003470:Culture Media; D011993:Recombinant Fusion Proteins; C017721:casamino acids; D006639:Histidine; D009320:Atrial Natriuretic Factor; D007930:Leucine; D000225:Adenine