Influence of pH on recombinant human growth hormone production by Pichia pastoris

被引:37
|
作者
Calik, Pinar [1 ,2 ]
Bayraktar, Eda [1 ]
Inankur, Bahar [1 ]
Soyaslan, Elif S. [1 ]
Sahin, Merve [1 ]
Taspinar, Hatice [2 ]
Acik, Eda [1 ]
Yilmaz, Remziye [2 ]
Ozdamar, Tuncer H. [3 ]
机构
[1] Middle E Tech Univ, Dept Chem Engn, Ind Biotechnol and Metab Engn Lab, TR-06531 Ankara, Turkey
[2] Middle E Tech Univ, Dept Biotechnol, Grad Sch Nat & Appl Sci, TR-06531 Ankara, Turkey
[3] Ankara Univ, Dept Chem Engn, Biochem React Engn Lab, TR-06100 Ankara, Turkey
关键词
Pichia pastoris; pH; sorbitol; gene expression; AOX; human growth hormone; BACILLUS-LICHENIFORMIS; STRUCTURAL-ANALYSIS; PROTEASE PRODUCTION; EXPRESSION SYSTEM; FERMENTATION; YEAST; BATCH; SECRETION; PURIFICATION; PROTEINS;
D O I
10.1002/jctb.2474
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
BACKGROUND: Effect of pH on recombinant human growth hormone (rhGH) production by Pichia pastoris hGH-Mut(+) was investigated at pH = 4.2, 5.0, 5.5, and 6.0. RESULTS: The highest cell concentration was obtained at pH = 6.0 with 53 g L(-1), while the highest rhGH concentration was attained at pH = 5.0 as 0.27 g L(-1). Total protease secretion increased with increase in pH and with the cultivation time. Oxygen uptake rate increased with increasing pH up to pH = 6.0, having the maximum value, 37 mmol m(-3) s(-1), at pH= 5.5. K(L)a values were similar at all the conditions, having a maximum value of 0.14 s(-1) at pH = 5.0. Taking the final rhGH concentration into account, the most favourable pH was 5.0; where AOX1 expression level showed a similar trend to AOX activity profiles, having the highest value of 9.4 x 10(10) copy mg(-1) CDW at t = 15 h; in parallel to AOX1 expression profile, hGH expression level increased until t = 15 h, with the highest value of 4.0 x 10(10) copy mg(-1) CDW, where a sharp increase in rhGH concentration was obtained. The expression levels of pep4, prb1 and prc1 genes, responsible from the production of proteinase A, proteinase B and, carboxypeptidase Y, were parallel to each other. CONCLUSION: Since it was shown that pH is a crucial operating parameter in fermentation processes using P. pastoris, keeping pH constant at its determined optimum value, pH = 5.0, during the bioprocess is vital in terms of recombinant protein production. (C) 2010 Society of Chemical Industry
引用
收藏
页码:1628 / 1635
页数:8
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