The Kluyveromyces lactis Cu/Zn SOD gene (SOD1) was fused with the toxin K1 signal sequence to obtain extracellular production of superoxide dismutase. Kluyveromyces marxianus L3 and K. lactis MW98-8C strains were transformed and compared as hosts for the secretion. The effects of the media composition were evaluated: In K. lactis, the highest volumetric activity was obtained in YKK synthetic medium in the presence of Cu(2+)/Zn(2+) cofactors (9.6 kU l(-1)). In K. marxianus, active SOD was produced only in YPD medium supplemented with Cu(2+) and Zn(2+) (8.8 kU l(-1)). In order to improve the production of secreted active SOD in K. lactis, the SOD1 copper carrier (CCS1) was overexpressed and targeted to the secretory apparatus. A positive effect was observed only when K. lactis was grown in a medium without Cu(2+)/Zn(2+) supplement. The best performing culture conditions for K. lactis and K. marxianus recombinant strains were successfully applied to two laboratory-scale fed-batch processes, and volumetric SOD activities increased up to 19.4 and 24.1 kU l(-1), respectively.

Secretion of Kluyveromyces lactis Cu/Zn SOD: strategies for enhanced production / Raimondi, Stefano; D., Uccelletti; Amaretti, Alberto; Leonardi, Alan; C., Palleschi; Rossi, Maddalena. - In: APPLIED MICROBIOLOGY AND BIOTECHNOLOGY. - ISSN 1432-0614. - ELETTRONICO. - 86(3):(2010), pp. 871-878. [10.1007/s00253-009-2353-5]

Secretion of Kluyveromyces lactis Cu/Zn SOD: strategies for enhanced production

RAIMONDI, Stefano;AMARETTI, Alberto;LEONARDI, Alan;ROSSI, Maddalena
2010

Abstract

The Kluyveromyces lactis Cu/Zn SOD gene (SOD1) was fused with the toxin K1 signal sequence to obtain extracellular production of superoxide dismutase. Kluyveromyces marxianus L3 and K. lactis MW98-8C strains were transformed and compared as hosts for the secretion. The effects of the media composition were evaluated: In K. lactis, the highest volumetric activity was obtained in YKK synthetic medium in the presence of Cu(2+)/Zn(2+) cofactors (9.6 kU l(-1)). In K. marxianus, active SOD was produced only in YPD medium supplemented with Cu(2+) and Zn(2+) (8.8 kU l(-1)). In order to improve the production of secreted active SOD in K. lactis, the SOD1 copper carrier (CCS1) was overexpressed and targeted to the secretory apparatus. A positive effect was observed only when K. lactis was grown in a medium without Cu(2+)/Zn(2+) supplement. The best performing culture conditions for K. lactis and K. marxianus recombinant strains were successfully applied to two laboratory-scale fed-batch processes, and volumetric SOD activities increased up to 19.4 and 24.1 kU l(-1), respectively.
86(3)
871
878
Secretion of Kluyveromyces lactis Cu/Zn SOD: strategies for enhanced production / Raimondi, Stefano; D., Uccelletti; Amaretti, Alberto; Leonardi, Alan; C., Palleschi; Rossi, Maddalena. - In: APPLIED MICROBIOLOGY AND BIOTECHNOLOGY. - ISSN 1432-0614. - ELETTRONICO. - 86(3):(2010), pp. 871-878. [10.1007/s00253-009-2353-5]
Raimondi, Stefano; D., Uccelletti; Amaretti, Alberto; Leonardi, Alan; C., Palleschi; Rossi, Maddalena
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Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/11380/640773
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