Heterologous production of Rhizopus oryzae lipase in Pichia pastoris using the alcohol oxidase and formaldehyde dehydrogenase promoters in batch and fed-batch cultures

A Rhizopus oryzae lipase (ROL) has been expressed in Pichia pastoris as a reporter gene using two different regulated promoters. Both phenotypes, Mut(s) and Mut(+), have been used for the expression under the control of alcohol oxidase I promoter (PAOX1). Moreover, the new formaldehyde dehydrogenase I promoter (PFLD1) has been tested. PFLD1 allows the design of methanol-free culture strategies, being methylamine a less volatile and flammable inducer. The comparison in batch and fed-batch cultures of the two promoters has shown that Mut(s) (PAOX1) is somewhat more efficient than PFLD1] and Mut(+) (PAOX1) systems, with a 1.1- and 1.4-fold higher specific productivity (U lipase/g biomass h), respectively. However, the productivity in all three systems was very similar (2500 U lipase/1 h). These results demonstrate that the expression of heterologous proteins under the transcriptional control of PFLD1 is a promising alternative to the well-known PAOX1. (c) 2005 Elsevier B.V. All rights reserved.