TY - JOUR
T1 - Searching the best operational strategies for Rhizopus oryzae lipase production in Pichia pastoris Mut+ phenotype: Methanol limited or methanol non-limited fed-batch cultures?
AU - Barrigón, José Manuel
AU - Montesinos, José Luis
AU - Valero, Francisco
PY - 2013/6/5
Y1 - 2013/6/5
N2 - Two different operational strategies, methanol limited (MLFB) and methanol non-limited fed-batch (MNLFB) cultures, have been compared for the production of the recombinant Rhizopus oryzae lipase (ROL) expressed in Pichia pastoris Mut+ phenotype. Yields, productivities and specific production rate in all MLFB conditions were very low, obtaining the best results at the lower specific growth rates. Similar results were obtained for MNLFB strategy at methanol set-point concentrations up to 2gl-1. However, for methanol set-points higher than 2gl-1 a significant increase in the production was observed. Methanol set-point of 3gl-1 was the optimum to maximize product yield (YP/X), both volumetric and specific productivity and mean specific production rate (qP,mean), although in terms of ROL production high values were obtained in the range of 3-10gl-1. An inhibitory effect of methanol on cell growth as well as slowness on ROL production during early induction phase was observed at concentration of 10gl-1. © 2013 Elsevier B.V.
AB - Two different operational strategies, methanol limited (MLFB) and methanol non-limited fed-batch (MNLFB) cultures, have been compared for the production of the recombinant Rhizopus oryzae lipase (ROL) expressed in Pichia pastoris Mut+ phenotype. Yields, productivities and specific production rate in all MLFB conditions were very low, obtaining the best results at the lower specific growth rates. Similar results were obtained for MNLFB strategy at methanol set-point concentrations up to 2gl-1. However, for methanol set-points higher than 2gl-1 a significant increase in the production was observed. Methanol set-point of 3gl-1 was the optimum to maximize product yield (YP/X), both volumetric and specific productivity and mean specific production rate (qP,mean), although in terms of ROL production high values were obtained in the range of 3-10gl-1. An inhibitory effect of methanol on cell growth as well as slowness on ROL production during early induction phase was observed at concentration of 10gl-1. © 2013 Elsevier B.V.
KW - Fed-batch culture
KW - Fermentation
KW - P AOX1
KW - Pichia pastoris
KW - Recombinant protein production
KW - Rhizopus oryzae lipase
U2 - 10.1016/j.bej.2013.03.018
DO - 10.1016/j.bej.2013.03.018
M3 - Article
SN - 1369-703X
VL - 75
SP - 47
EP - 54
JO - Biochemical Engineering Journal
JF - Biochemical Engineering Journal
ER -