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Abstract

Methylotrophic yeasts such as Candida boidinii, Hansenula polymorpha, Pichia methanolica and Pichia pastoris are an emerging group of eukaryotic hosts for recombinant protein production with an ever increasing number of applications during the last 30 years. Their applications are linked to the use of strong methanol-inducible promoters derived from genes of the methanol utilisation pathway. These promoters are tightly regulated, highly repressed in presence of non-limiting concentrations of glucose in the medium and strongly induced if methanol is used as carbon source. Several factors involved in this tight control and their regulatory effects have been described so far. This review summarises available data about the regulation of promoters from methanol utilisation pathway genes. Furthermore, the role of cis and trans acting factors (e.g. transcription factors, glucose processing enzymes) in the expression of methanol utilisation pathway genes is reviewed both in the context of the native cell environment as well as in heterologous hosts.

Superoxide Dismutase Enzymes in Oxidative Type YeastH. Polymorpha

“…The in silico study of Sod2p revealed that at the N-terminal 20 residues of the protein there is a positively charged region and at 35 position a R-2 motif RRT|KV was found ("Gavel" cleavage site), which suggested the mitochondrial localization of the protein. These results coincided with already well known data about mitochondrial matrix localization of this enzyme and its key role in protecting these organelles from free radicals damage (4,12,23). Furthermore, the enzyme possess possible PTS1 signal -GKI which could be responsible for its targeting to yeast peroxisomes.…”

Molecular Characterization of Hap Complex Components Responsible for Methanol-Inducible Gene Expression in the Methylotrophic Yeast Candida boidinii

“…Efficient production has been achieved with their strong and tightly regulated methanolinducible gene promoters in combination with high-cell-density cultures in methanol-containing medium. The genes encoding methanol-metabolic enzymes are highly induced in the presence of methanol, i.e., alcohol oxidase (AOD), dihydroxyacetone synthase (DAS), glutathione-dependent formaldehyde dehydrogenase (FLD), and formate dehydrogenase (FDH), and these promoter regions have been used commercially for heterologous gene expression (5)(6)(7).…”

Efficient Biocatalytic Synthesis of (R)‐Pantolactone

“…In order to boost the biocatalytic system from another side, in parallel, a synthetic Pahnl5 V317A gene was designed and expressed with the help of the improved promoter variant P AOX1D1 [21,22] and the co-expression of PDI. [23] Thus, a further 4-fold improved enzyme activity per volume of yeast culture was obtained.…”