Dihydroxyacetone synthase is localized in the peroxisomal matrix of methanol-grown Hansenula polymorpha

Douma, A.C.; Veenhuis, Marten; Koning, Willem de; Evers, Melchior E.; Harder, W.

doi:10.1007/bf00411242

Cited by 154 publications

(151 citation statements)

References 29 publications

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“…Enzymatic activities of the peroxisomal marker catalase (Luck, 1963) and of the mitochondrial marker cytochrome c oxidase (Douma et al, 1985) were measured by established procedures. SDS-PAGE (Laemmli, 1970) and immunoblotting with the use of semidry electrophoretic transfer (Kyhse-Andersen, 1984) were performed as described.…”

Section: Analytical Proceduresmentioning

confidence: 99%

Mutants of theYarrowia lipolytica PEX23Gene Encoding an Integral Peroxisomal Membrane Peroxin Mislocalize Matrix Proteins and Accumulate Vesicles Containing Peroxisomal Matrix and Membrane Proteins

Brown

Titorenko

Rachubinski

2000

MBoC

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pex mutants are defective in peroxisome assembly. The mutant strain pex23-1 of the yeast Yarrowia lipolytica lacks morphologically recognizable peroxisomes and mislocalizes all peroxisomal matrix proteins investigated preferentially to the cytosol. pex23 strains accumulate vesicular structures containing both peroxisomal matrix and membrane proteins. The PEX23 gene was isolated by functional complementation of the pex23-1 strain and encodes a protein, Pex23p, of 418 amino acids (47,588 Da). Pex23p exhibits high sequence similarity to two hypothetical proteins of the yeastSaccharomyces cerevisiae. Pex23p is an integral membrane protein of peroxisomes that is completely, or nearly completely, sequestered from the cytosol. Pex23p is detected at low levels in cells grown in medium containing glucose, and its levels are significantly increased by growth in medium containing oleic acid, the metabolism of which requires intact peroxisomes.

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Section: Analytical Proceduresmentioning

confidence: 99%

Mutants of theYarrowia lipolytica PEX23Gene Encoding an Integral Peroxisomal Membrane Peroxin Mislocalize Matrix Proteins and Accumulate Vesicles Containing Peroxisomal Matrix and Membrane Proteins

Brown

Titorenko

Rachubinski

2000

MBoC

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“…Electron microscopy and immunogoldlabeling of NAD-dependent methanol dehydrogenase were performed as described by Douma et al (1985).…”

Section: Electron Microscopy and Immunocytochemistrymentioning

confidence: 99%

Methanol metabolism in thermotolerant methylotrophic Bacillus strains involving a novel catabolic NAD-dependent methanol dehydrogenase as a key enzyme

et al. 1989

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Abstract. The enzymology of methanol utilization in thermotolerant methytotrophic Bacillus strains was investigated. In all strains an immunological]y related NAD-dependent methanol dehydrogenase was involved in the initial oxidation of methanol. In cells of Bacillus sp. C1 grown under methanol-limiting conditions this enzyme constituted a high percentage of total soluble protein. The methanol dehydrogenase from this organism was purified to homogeneity and characterized. In cell-free extracts the enzyme displayed biphasic kinetics towards methanol, with apparent Km values of 3.8 and 166 mM. Carbon assimilation was by way of the fructose-1,6-bisphosphate aldolase cleavage and transketolase/transaldolase rearrangement variant of the RuMP cycle of formaldehyde fixation. The key enzymes of the RuMP cycle, hexulose-6-phosphate synthase (HPS) and hexulose-6-phosphate isomerase (HPI), were present at very high levels of activity. Failure of whole cells to oxidize formate, and the absence of formaldehyde-and formate dehydrogenases indicated the operation of a non-linear oxidation sequence for formaldehyde via HPS. A comparison of the levels of methanol dehydrogenase and HPS in cells of Bacillus sp. C1 grown on methanol and glucose suggested that the synthesis of these enzymes is not under coordinate control.

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“…For transfer of ceils from methanol to ethanol-or glucose-containing media, H. polymorpha was cultivated in a continuous culture at a dilution rate of 0.06 h-1 in the medium described by Douma et al (1985) which contained 0.5% (w/v) methanol as the carbon source. …”

Section: Organandms and Growth Conditionsmentioning

confidence: 99%

“…Protoplasts from H. polymorpha were prepared as described by Douma et al (1985), from C. utilis as described by Zwart et al (1983) and from T. cutaneum X4 as described by .…”

Section: Formation Of Protoplastsmentioning

confidence: 99%

A 31P NMR study of the internal pH of yeast peroxisomes

et al. 1987

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Dihydroxyacetone synthase is localized in the peroxisomal matrix of methanol-grown Hansenula polymorpha

Cited by 154 publications

References 29 publications

Mutants of theYarrowia lipolytica PEX23Gene Encoding an Integral Peroxisomal Membrane Peroxin Mislocalize Matrix Proteins and Accumulate Vesicles Containing Peroxisomal Matrix and Membrane Proteins

Mutants of theYarrowia lipolytica PEX23Gene Encoding an Integral Peroxisomal Membrane Peroxin Mislocalize Matrix Proteins and Accumulate Vesicles Containing Peroxisomal Matrix and Membrane Proteins

Methanol metabolism in thermotolerant methylotrophic Bacillus strains involving a novel catabolic NAD-dependent methanol dehydrogenase as a key enzyme

A 31P NMR study of the internal pH of yeast peroxisomes

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