Methanol metabolism in mutants of the methylotrophic yeast Hansenula polymorpha

Bystrykh, L

doi:10.1016/0378-1097(88)90378-3

Cited by 4 publications

(11 citation statements)

References 8 publications

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“…To confirm the involvement of FDH1 in methylamine-or choline-grown cells, we compared the wild-type strain and the fdh1⌬ mutant for growth on methylamine or choline. As described above, although the growth of the fdh1⌬ mutant on methanol and NH 4 Cl as carbon and nitrogen sources, respectively, was inhibited, its growth on glucose-NH 4 Cl was not affected at all. The growth of the fdh1⌬ mutant on glucose-methylamine and glucose-choline media was slightly inhibited but not as severely as growth on methanol as a carbon source.…”

Section: Vol 179 1997 Regulation Of Formate Dehydrogenase In C Boimentioning

confidence: 67%

“…4A, lanes 1 and 14). On the other hand, when methylamine or choline was used as a single nitrogen source in place of NH 4 Cl, FDH activity was induced in glucose-or glycerol-grown cells (Fig. 4A, lanes 6 to 9).…”

Section: Vol 179 1997 Regulation Of Formate Dehydrogenase In C Boimentioning

confidence: 98%

“…However, a five-to eightfold higher level of formate (1.8 to 2.8 mM after 84 h) was detected in the culture medium of the fdh1⌬ strain than in that of the wild-type strain. When glucose-NH 4 Cl was used for the growth medium, formate was not detected. From these results, we assume that formate accumulation is the main reason for the growth defect of the fdh1⌬ mutant with methylamine or choline as a nitrogen source.…”

Section: Vol 179 1997 Regulation Of Formate Dehydrogenase In C Boimentioning

confidence: 99%

“…Among the carbon sources tested when NH 4 Cl was used as a single nitrogen source, only methanol gave a high level of FDH activity (Fig. 4A, lanes 1 to 5).…”

Section: Vol 179 1997 Regulation Of Formate Dehydrogenase In C Boimentioning

confidence: 99%

“…There is some discrepancy regarding the role of FDH in the methylotrophic growth of yeast cells. Bystrykh et al reported that an FDH-deficient mutant of H. polymorpha could not grow on methanol (4). On the other hand, Sibirny et al reported that neither FLD nor FDH was essential to the energy supply for the methylotrophic growth of H. polymorpha (26).…”

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Regulation of the formate dehydrogenase gene, FDH1, in the methylotrophic yeast Candida boidinii and growth characteristics of an FDH1-disrupted strain on methanol, methylamine, and choline

et al. 1997

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The structural gene (FDH1) coding for NAD ؉ -dependent formate dehydrogenase (FDH) was cloned from a genomic library of Candida boidinii, and the FDH1 gene was disrupted in the C. boidinii genome (fdh1⌬) by one-step gene disruption. In a batch culture experiment, although the fdh1⌬ strain was still able to grow on methanol, its growth was greatly inhibited and a toxic level of formate was detected in the medium. In a methanol-limited chemostat culture at a low dilution rate (0.03 to 0.05 h ؊1 ), formate was not detected in the culture medium of the fdh1⌬ strain; however, the fdh1⌬ strain showed only one-fourth of the growth yield of the wild-type strain. Expression of FDH1 was found to be induced by choline or methylamine (used as a nitrogen source), as well as by methanol (used as a carbon source). Induction of FDH1 was not repressed in the presence of glucose when cells were grown on methylamine, choline, or formate, and expression of FDH1 was shown to be regulated at the mRNA level. Growth on methylamine or choline as a nitrogen source in a batch culture was compared between the wild type and the fdh1⌬ mutant. Although the growth of the fdh1⌬ mutant was impaired and the level of formate was higher in the fdh1⌬ mutant than in the wild-type strain, the growth defect caused by FDH1 gene disruption was small and less severe than that caused by growth on methanol. As judged from these results, the main physiological role of FDH with all of the FDH1-inducing growth substrates seems to be detoxification of formate, and during growth on methanol, FDH seems to contribute significantly to the energy yield.

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Section: Vol 179 1997 Regulation Of Formate Dehydrogenase In C Boimentioning

confidence: 67%

Section: Vol 179 1997 Regulation Of Formate Dehydrogenase In C Boimentioning

confidence: 98%

Section: Vol 179 1997 Regulation Of Formate Dehydrogenase In C Boimentioning

confidence: 99%

“…Among the carbon sources tested when NH 4 Cl was used as a single nitrogen source, only methanol gave a high level of FDH activity (Fig. 4A, lanes 1 to 5).…”

Section: Vol 179 1997 Regulation Of Formate Dehydrogenase In C Boimentioning

confidence: 99%

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confidence: 99%

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Regulation of the formate dehydrogenase gene, FDH1, in the methylotrophic yeast Candida boidinii and growth characteristics of an FDH1-disrupted strain on methanol, methylamine, and choline

et al. 1997

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The NAD‐dependent alcohol dehydrogenase (ADH) of hansenula polymorpha cbs 4732 – an electrophoretic study (II)

Steudel

Nitzsche

1991

Acta Biotechnologica

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Using gradient polyacrylamide gel electrophoresis (gradient between 10-20%) we were able to show up to six clearly distinguishable ADH-bands in Hansenulu polymorphu CBS 4732. In contrast t o commercial ADH from Saccharomyces cerevisiae the ADH-forms of Hamenula polymorpha CBS 4732 have a much broader substrate specificity. Furthermore, the occurence of secondary alcohol dehydrogenases and of an aromatic alcohol dehydrogenase could be demonstrated.

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Physiological role of the glutathione-dependent formaldehyde dehydrogenase in the methylotrophic yeast Candida boidinii b bThe GenBank accession number for the sequence reported in this paper is AB085186.

et al. 2002

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The methylotrophic yeast Candida boidinii exhibits formaldehyde dehydrogenase activity (FLD, EC 1.2.1.1) during growth on methanol as a sole carbon source. The structural gene, FLD1, was cloned from a genomic library of C. boidinii. The 1263 bp FLD1 gene contained a 123 bp intron and its exon encoded a gene product of 380 amino acids, whose predicted amino acid sequence showed high similarity to the sequences of FLDs from other organisms. The FLD1 gene was disrupted in the C. boidinii genome by one-step gene disruption. The fld1∆ strain could not grow on methanol as a carbon source under methanol-limited chemostat culture conditions, even with low dilution rates (D T 005 h N1 ), whereas a strain with a disruption in the gene for formate dehydrogenase (FDH ; another NADH-generating dehydrogenase involved in the formaldehyde oxidation pathway) could survive. These results indicated that FLD, but not FDH, is essential for growth of C. boidinii on methanol.

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Methanol metabolism in mutants of the methylotrophic yeast Hansenula polymorpha

Cited by 4 publications

References 8 publications

Regulation of the formate dehydrogenase gene, FDH1, in the methylotrophic yeast Candida boidinii and growth characteristics of an FDH1-disrupted strain on methanol, methylamine, and choline

Regulation of the formate dehydrogenase gene, FDH1, in the methylotrophic yeast Candida boidinii and growth characteristics of an FDH1-disrupted strain on methanol, methylamine, and choline

The NAD‐dependent alcohol dehydrogenase (ADH) of hansenula polymorpha cbs 4732 – an electrophoretic study (II)

Physiological role of the glutathione-dependent formaldehyde dehydrogenase in the methylotrophic yeast Candida boidinii b bThe GenBank accession number for the sequence reported in this paper is AB085186.

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