Enhancement of Human γ-Interferon Production in Recombinant E-coli Using Batch Cultivation

被引:23
作者
Babaeipour, Valiollah [1 ]
Shojaosadati, Seyed Abbas [1 ]
Khalilzadeh, Rasoul [2 ]
Maghsoudi, Nader [3 ]
Farnoud, Amir Mohammad [1 ]
机构
[1] Univ Tarbiat Modares, Dept Chem Engn, Biotechnol Grp, Fac Engn, Tehran, Iran
[2] Biotechnol Res Ctr, Grp Biochem Engn, Tehran, Iran
[3] Med Sci Univ Shahid Beheshti, Neurosci Res Ctr, Tehran, Iran
关键词
gamma-Interferon; Induction condition; Productivity; E; coli; Batch culture; HIGH CELL-DENSITY; CULTURE-CONDITIONS; DEFINED MEDIA; EXPRESSION; PROTEINS; OPTIMIZATION; STRATEGIES; INDUCER; PROMOTER; LACTOSE;
D O I
10.1007/s12010-009-8718-5
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Development of inexpensive and simple culture media and appropriate induction conditions are always favorable for industry. In this research, chemical composition and stoichiometric data for gamma-interferon production and recombinant Escherichia coli growth were used in order to achieve a simple medium and favorable induction conditions. To achieve this goal, the effects of medium composition and induction conditions on the production of gamma-interferon were investigated in batch culture of E. coli BL21 (DE3) [pET3a-ifn gamma]. These conditions were considered as suitable conditions for the production of gamma-interferon: 2.5x M9 medium, supplemented with a mixture of amino acids (milligram per liter), including glutamic acid 215, aspartic acid 250, lysine 160, and phenylalanine 90, and induction at late-log phase (OD600=4.5). Under these conditions, dry cell weight of 6 +/- 0.2 g/l and gamma-interferon concentration of 2.15 +/- 0.1 g/l were obtained. Later, without changing the concentration ratio of amino acids and glucose, the effect of increase in the primary glucose concentration on productivity of gamma-interferon was investigated. It was found that 25 g/l glucose will result in maximum attainable biomass and recombinant human gamma-interferon. At improved conditions, a dry cell weight of 14 +/- 0.2 g/l, concentration and overall productivity of gamma-interferon 4.2 +/- 0.1 g/l and 420 +/- 10 mg/l h, respectively, were obtained.
引用
收藏
页码:2366 / 2376
页数:11
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