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Updated: Jun 4, 2026
Methanol Independent Expression by Pichia Pastoris Employing De-repression Technologies
Published on: January 23, 2019
Molecular characterization of the PEX14 gene from the methylotrophic yeast Pichia methanolica
Takashi Ito1, Daisuke Ito, Shotaro Ozawa
1Department of Food Science and Technology, Faculty of Bioindustry, Tokyo University of Agriculture, 196 Yasaka, Abashiri, Hokkaido 099-2493, Japan.
Abstract:
In this study, we describe the molecular characterization of the PmPEX14 gene encoding the peroxisomal membrane protein from the methylotrophic yeast Pichia methanolica. The pex14Δ strain of P. methanolica lost its ability to grow on methanol and oleate but grew normally on glucose. Disruption of the PmPEX14 caused a decrease in the activities of peroxisomal methanol-metabolizing enzymes and mislocalization of those proteins into the cytosol and vacuole. Taken together, these findings show that PmPex14p has an essential physiological role in methanol metabolism in P. methanolica.
Insights
The PmPEX14 gene is crucial for Pichia methanolica to grow on methanol. Its disruption impairs methanol metabolism by mislocalizing key enzymes.
Area of Science:
- Microbiology
- Molecular Biology
- Biochemistry
Background:
- Methylotrophic yeasts like Pichia methanolica utilize methanol as a carbon source.
- Peroxisomes are essential organelles for methanol metabolism in these yeasts.
- The PEX14 gene encodes a peroxisomal membrane protein involved in peroxisome biogenesis and function.
Purpose of the Study:
- To investigate the molecular characterization and function of the PmPEX14 gene in Pichia methanolica.
- To determine the role of PmPex14p in methanol metabolism and peroxisomal protein targeting.
Main Methods:
- Molecular cloning and gene disruption of PmPEX14 in P. methanolica.
- Growth assays on various carbon sources (glucose, methanol, oleate).
- Enzyme activity assays for methanol-metabolizing enzymes.
- Subcellular localization studies of peroxisomal proteins using microscopy.
Main Results:
- Disruption of PmPEX14 resulted in the inability of P. methanolica to grow on methanol and oleate.
- The pex14Δ mutant showed reduced activity of peroxisomal methanol-metabolizing enzymes.
- Methanol-metabolizing enzymes were mislocalized from peroxisomes to the cytosol and vacuole in the pex14Δ strain.
Conclusions:
- PmPEX14 plays an essential physiological role in methanol metabolism in Pichia methanolica.
- PmPex14p is critical for the proper targeting and function of methanol-metabolizing enzymes within peroxisomes.
