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

• Published in: Journal of biotechnology 2007-06, Vol.130 (3), p.291-299
• Main Authors: Surribas, Anna, Stahn, Rainer, Montesinos, José Luis, Enfors, Sven-Olof, Valero, Francisco, Jahic, Mehmedalija
• Format: Article
• Language: English
• Subjects: Alcohol Oxidoreductases - metabolism; batch systems; Biological and medical sciences; Biomass; Bioreactors; biosynthesis; Biotechnology; Cell Culture Techniques - methods; Cell death; cultivation; culture media; expression; fed-batch culture; fermentation; Fermentation - drug effects; Fundamental and applied biological sciences. Psychology; gene expression; gene transfer; genetic transformation; growth; heterologous protein-production; high-level production; Lipase - biosynthesis; Lipolysis - drug effects; methanol; Methanol - pharmacology; Microbiological Techniques; Operational fed-batch strategies; Pichia - cytology; Pichia - drug effects; Pichia - metabolism; Pichia pastoris; proteolytic degradation; recombinant protein; Rhizopus - drug effects; Rhizopus - enzymology; Rhizopus orysae lipase; Rhizopus oryzae; Rhizopus oryzae lipase; salt content; Salts - pharmacology; Temperature; Time Factors; triacylglycerol lipase; viability
• ISSN: 0168-1656; 1873-4863; 1873-4863
• DOI: 10.1016/j.jbiotec.2007.04.009

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.