Production of a Rhizopus oryzae lipase from Pichia pastoris using alternative operational strategies.
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.
Surribas
Anna
A
Departament d'Enginyeria Química, Escola Tècnica Superior d'Enginyeria, Universitat Autònoma de Barcelona, 08193 Bellaterra, Barcelona, Spain.
Stahn
Rainer
R
Montesinos
José Luis
JL
Enfors
Sven-Olof
SO
Valero
Francisco
F
Jahic
Mehmedalija
M
D000429:Alcohol Oxidoreductases / Q000378:metabolism; D018533:Biomass; D019149:Bioreactors; D018929:Cell Culture Techniques / Q000379:methods*; D005285:Fermentation / Q000187:drug effects; D008049:Lipase / Q000096:biosynthesis*; D008066:Lipolysis / Q000187:drug effects; D000432:Methanol / Q000494:pharmacology; D008828:Microbiological Techniques; D010843:Pichia / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D012233:Rhizopus / Q000187:drug effects / Q000201:enzymology*; D012492:Salts / Q000494:pharmacology; D013696:Temperature; D013997:Time Factors
D012492:Salts; D000429:Alcohol Oxidoreductases; C019643:alcohol oxidase; D008049:Lipase; D000432:Methanol