Production of a Rhizopus oryzae lipase from Pichia pastoris using alternative operational strategies

被引:41
|
作者
Surribas, Anna
Stahn, Rainer
Montesinos, Jose Luis
Enfors, Sven-Olof
Valero, Francisco [1 ]
Jahic, Mehmedalija
机构
[1] Univ Autonoma Barcelona, Escola Tecn Super Engn, Dept Engn Quim, Bellaterra 08193, Spain
[2] Royal Inst Technol, Dept Biotechnol, S-10691 Stockholm, Sweden
关键词
Pichia pastoris; operational fed-batch strategies; Rhizopus orysae lipase; cell death; HETEROLOGOUS PROTEIN-PRODUCTION; HIGH-LEVEL PRODUCTION; RECOMBINANT PROTEIN; PROTEOLYTIC DEGRADATION; EXPRESSION; METHANOL; GROWTH; FERMENTATION; CULTIVATION; VIABILITY;
D O I
10.1016/j.jbiotec.2007.04.009
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
Different cultivation strategies have been compared for the production of Rhizopus oryzae lipase (ROL) from Pichia pastoris. Several drawbacks have been found using a methanol non-limited fed-batch. On the one hand, oxygen limitation appeared at early cell dry weights and, on the other hand, high cell death was observed. A temperature limited fed-batch has been proposed to solve both problems. However, in our case study a methanol non-limited fed-batch results in better productivities. Finally, a lower salt medium were used to overcome cell death problems and a temperature limited fed-batch was applied thereafter to solve oxygen transfer limitations. This combined strategy has resulted in lower productivities when compared to a methanol non-limited fed-batch. However the culture could be longer prolonged and a 1.3-fold purer final product was obtained mainly due to cell death reduction. (c) 2007 Elsevier B.V. All rights reserved.
引用
收藏
页码:291 / 299
页数:9
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