Methanol-Utilizing Yeasts

Koning, Willem de; Harder, W.

doi:10.1007/978-1-4899-2338-7_7

Cited by 21 publications

(11 citation statements)

References 96 publications

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“…In this pathway, methanol is first oxidized to formaldehyde. To this one-carbon compound a two-carbon hydroxyethyl group is then transferred from xylulose-5-phosphate by Dha-synthase, a thiamine pyrophosphate-dependent transketolase [16][17][18][19][20][21]. Bacteria, for comparison, utilize other routes to assimilate formaldehyde and its oxidation product CO 2 , namely the ribulose monophosphate (RuMP) cycle, the serine pathway and the reductive pentose phosphate cycle [21,22].…”

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confidence: 99%

Small substrate, big surprise: fold, function and phylogeny of dihydroxyacetone kinases

Erni

Siebold

Christen

et al. 2006

Cell. Mol. Life Sci.

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Dihydroxyacetone (Dha) kinases are a family of sequence-conserved enzymes which utilize either ATP (in animals, plants and eubacteria) or phosphoenolpyruvate (PEP, in eubacteria) as their source of high-energy phosphate. The kinases consist of two domains/subunits: DhaK, which binds Dha covalently in hemiaminal linkage to the Nepsilon2 of a histidine, and DhaL, an eight-helix barrel that contains the nucleotide-binding site. The PEP-dependent kinases comprise a third subunit, DhaM, which rephosphorylates in situ the firmly bound ADP cofactor. DhaM serves as the shuttle for the transfer of phosphate from the bacterial PEP: carbohydrate phosphotransferase system (PTS) to the Dha kinase. The DhaL and DhaK subunits of the PEP-dependent Escherichia coli kinase act as coactivator and corepressor of DhaR, a transcription factor from the AAA(+) family of enhancerbinding proteins. In Gram-positive bacteria genes for homologs of DhaK and DhaL occur in operons for putative transcription factors of the TetR and DeoR families. Proteins with the Dha kinase fold can be classified into three families according to phylogeny and function: Dha kinases, DhaK and DhaL homologs (paralogs) associated with putative transcription regulators of the TetR and DeoR families, and proteins with a circularly permuted domain order that belong to the DegV family.

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confidence: 99%

Small substrate, big surprise: fold, function and phylogeny of dihydroxyacetone kinases

Erni

Siebold

Christen

et al. 2006

Cell. Mol. Life Sci.

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“…It is generally known that several methylotrophic bacteria contain key enzymes of more than one methanol assimilation pathway, though not the yeast xylulose monophosphate (XuMP) pathway, and no methylotrophic yeast possesses the key enzymes for prokaryotic C 1 assimilation pathways such as the ribulose monophosphate (RuMP) and serine pathways and the Calvin cycle (Anthony, 1982;de Koning & Harder, 1992;Dijkhuizen et al, 1992; Fig. 1).…”

Section: Introductionmentioning

confidence: 99%

“…strain JC1 has been found to begin with the XuMP pathway, followed by both the XuMP pathway and the Calvin cycle (Ro et al, 1997a). DHAS (EC 2.2.1.3) is the key enzyme responsible for formaldehyde fixation in methylotrophic organisms adapting to the XuMP pathway during growth on methanol as a sole source of carbon and energy (Anthony, 1982;de Koning & Harder, 1992;Dijkhuizen et al, 1992). The enzyme has been regarded as a special transketolase (TKT) because it is able to catalyse the transfer of glycoaldehyde from the donor xylulose 5-phosphate to the acceptor formaldehyde, the first intermediate produced during methanol oxidation ( Fig.…”

Section: Introductionmentioning

confidence: 99%

Cloning, characterization and expression of a gene encoding dihydroxyacetone synthase in Mycobacterium sp. strain JC1 DSM 3803

Seo

Park

et al. 2007

Microbiology

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Dihydroxyacetone synthase (DHAS) is a key enzyme involved in the assimilation of methanol in Mycobacterium sp. strain JC1 DSM 3803. The structural gene encoding DHAS in Mycobacterium sp. strain JC1 was cloned using random-primed probes synthesized after PCR with synthetic primers based on the amino acid sequences conserved in two yeast DHASs and several transketolases. The cloned gene, dasS, had an ORF of 2193 nt, encoding a protein with a calculated molecular mass of 78 197 Da. The deduced amino acid sequence of dasS contained an internal sequence of Mycobacterium sp. strain JC1 DHAS and exhibited 29.2 and 27.3 % identity with those of Candida boidinii and Hansenula polymorpha enzymes, respectively. Escherichia coli transformed with the cloned gene produced a novel protein with a molecular mass of~78 kDa, which cross-reacted with anti-DHAS antiserum and exhibited DHAS activity. Primer-extension analysis revealed that the transcriptional start site of the gene was the nucleotide A located 31 bp upstream from the dasS start codon. RT-PCR showed that dasS was transcribed as a monocistronic message. Northern hybridization and b-galactosidase assay with the putative promoter region of dasS revealed that the gene was transcribed only in cells growing on methanol. The expression of dasS in Mycobacterium sp. strain JC1 was free from catabolite repression.

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“…The ribulose monophosphate (RuMP) cycle for the assimilation of formaldehyde, the first metabolite of methanol oxidation, the serine pathway for the fixation of one molecule each of formaldehyde and CO 2 , the end product of methanol oxidation, and the Calvin reductive pentose phosphate cycle for CO 2 fixation were found to work in methylotrophic bacteria, whereas the xylulose monophosphate (XuMP) cycle for the assimilation of formaldehyde was found only in methylotrophic yeasts (2,8,9).…”

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“…strain JC1 DSM 3803; i.e., the bacterium grown on methanol was found to exhibit activities of key enzymes for the Calvin and XuMP cycles but no activity of that for the RuMP cycle and very low activity of the serine pathway enzyme. We also report several properties of dihydroxyacetone synthase (DHAS), the key enzyme for XuMP cycle (2,8,9), purified from cells of Acinetobacter sp. strain JC1 DSM 3803 to learn more about the diversity of DHAS in methylotrophic organisms.…”

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Dihydroxyacetone synthase from a methanol-utilizing carboxydobacterium, Acinetobacter sp. strain JC1 DSM 3803

Eom

Song

et al. 1997

J Bacteriol

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Acinetobacter sp. strain JC1 DSM 3803, a carboxydobacterium, grown on methanol was found to show dihydroxyacetone synthase, dihydroxyacetone kinase, and ribulose 1,5-bisphosphate carboxylase, but no hydroxypyruvate reductase and very low hexulose 6-phosphate synthase, activities. The dihydroxyacetone synthase was found to be expressed earlier than the ribulose 1,5-bisphosphate carboxylase. The dihydroxyacetone synthase was purified 19-fold in eight steps to homogeneity, with a yield of 9%. The final specific activity of the purified enzyme was 1.12 mol of NADH oxidized per min per mg of protein. The molecular weight of the native enzyme was determined to be 140,000. Sodium dodecyl sulfate-gel electrophoresis revealed a subunit of molecular weight 73,000. The optimum temperature and pH were 30°C and 7.0, respectively. The enzyme was inactivated very rapidly at 70°C. The enzyme required Mg 2؉ and thiamine pyrophosphate for maximal activity. Xylulose 5-phosphate was found to be the best substrate when formaldehyde was used as a glycoaldehyde acceptor. Erythrose 4-phosphate, glycolaldehyde, and formaldehyde were found to act as excellent substrates when xylulose 5-phosphate was used as a glycoaldehyde donor. The K m s for formaldehyde and xylulose 5-phosphate were 1.86 mM and 33.3 M, respectively. The enzyme produced dihydroxyacetone from formaldehyde and xylulose 5-phosphate. The enzyme was found to be expressed only in cells grown on methanol and shared no immunological properties with the yeast dihydroxyacetone synthase.Carboxydobacteria are a group of facultative chemolithoautotrophic bacteria, except for Streptomyces thermoautotrophicus, which are able to grow aerobically at the expense of carbon monoxide (CO), in addition to several organic materials, as a sole source of carbon and energy (15,24). It has been known for a long time that Pseudomonas gazotropha is the only carboxydobacterium also capable of growing methylotrophically on methanol as the sole carbon and energy sources and is recognized as the first organism adopting three nutrition types, i.e., organotrophy, autotrophy, and methylotrophy (15,25,27,28,39). We, however, observed in a previous study that Acinetobacter sp. strain JC1 DSM 3803, a carboxydobacterium isolated in Korea (6), also grows methylotrophically on both methanol and methylamine as sole carbon and energy sources.It is well known that there are four different ways for assimilation of C 1 compounds in methylotrophic organisms (2). The ribulose monophosphate (RuMP) cycle for the assimilation of formaldehyde, the first metabolite of methanol oxidation, the serine pathway for the fixation of one molecule each of formaldehyde and CO 2 , the end product of methanol oxidation, and the Calvin reductive pentose phosphate cycle for CO 2 fixation were found to work in methylotrophic bacteria, whereas the xylulose monophosphate (XuMP) cycle for the assimilation of formaldehyde was found only in methylotrophic yeasts (2,8,9).In this study, we report the presence of a novel mechanism for the assim...

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Methanol-Utilizing Yeasts

Cited by 21 publications

References 96 publications

Small substrate, big surprise: fold, function and phylogeny of dihydroxyacetone kinases

Small substrate, big surprise: fold, function and phylogeny of dihydroxyacetone kinases

Cloning, characterization and expression of a gene encoding dihydroxyacetone synthase in Mycobacterium sp. strain JC1 DSM 3803

Dihydroxyacetone synthase from a methanol-utilizing carboxydobacterium, Acinetobacter sp. strain JC1 DSM 3803

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