Effect of oxygen supply on passaging, stabilising and screening of recombinant Hansenula polymorpha production strains in test tube cultures.
Twenty-four Hansenula polymorpha transformants were passaged and stabilised in glucose medium and screened in glycerol medium for recombinant phytase in shaken test tubes. The cultivations were performed under either limited or non-limited oxygen supply. Maximum oxygen transfer capacities of test tubes were assessed by sulfite oxidation. Oxygen-limited glucose cultures resulted in a partially anaerobic metabolism and formation of 4.1 g ethanol l(-1), which was subsequently aerobically metabolised. Non-limited oxygen supply led to overflow metabolism and to accumulation of 2.1 g acetic acid l(-1), reducing the biomass yield. The use of glycerol in the screening main cultures prevented by-product formation irrespective of oxygen supply. Preculturing in glucose medium under non-limited oxygen supply resulted in a 20-h lag phase of the screening main culture. This lag phase was not observed when preculturing was performed under oxygen limitation. Phytase activity was on average 25% higher in cultures passaged, stabilised and screened under limited oxygen supply than in cultures under non-limited oxygen supply.
Stöckmann
Christoph
C
Department of Biochemical Engineering, RWTH Aachen University, Sammelbau Biologie, Worringerweg 1, D-52056, Aachen, Germany.
Losen
Mario
M
Dahlems
Ulrike
U
Knocke
Christof
C
Gellissen
Gerd
G
Büchs
Jochen
J
D010832:6-Phytase / Q000378:metabolism*; D019342:Acetic Acid / Q000378:metabolism; D000332:Aerobiosis; D000693:Anaerobiosis; D003470:Culture Media / Q000378:metabolism; D000431:Ethanol / Q000378:metabolism; D005285:Fermentation; D005947:Glucose; D005990:Glycerol; D008828:Microbiological Techniques; D010100:Oxygen; D010843:Pichia / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*
D003470:Culture Media; D000431:Ethanol; D010832:6-Phytase; D005947:Glucose; D005990:Glycerol; D019342:Acetic Acid; D010100:Oxygen