The Experts below are selected from a list of 1719 Experts worldwide ranked by ideXlab platform
Yasuyoshi Sakai - One of the best experts on this subject based on the ideXlab platform.
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Msn5p is involved in formaldehyde resistance but not in oxidative stress response in the methylotrophic yeast Candida Boidinii.
Bioscience biotechnology and biochemistry, 2012Co-Authors: Zhenyu Zhai, Hiroya Yurimoto, Yasuyoshi SakaiAbstract:Methylotrophic yeasts, which can utilize methanol as sole carbon and energy source, are exposed to two toxic metabolic intermediates, formaldehyde and hydrogen peroxide, during growth on methanol. Here we report that Msn5p, an importin-β family nuclear exporter, participated in the formaldehyde resistance mechanism but not in the hydrogen peroxide resistance mechanism in Candida Boidinii. Disruption of the MSN5 gene in this yeast caused retardation of growth on formaldehyde-generating growth substrates such as methanol and methylamine, but the expression levels of the methanol-metabolizing enzymes did not fall. The Msn5p-depleted strain was sensitive to formaldehyde but not to hydrogen peroxide. Furthermore, a yellow fluorescent protein-tagged Msn5p was diffuse in the cytoplasm of C. Boidinii when the cells were treated with high concentrations of formaldehyde or ethanol, but was predominantly associated with the nuclei following treatment with hydrogen peroxide.
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Methanol-inducible gene expression and heterologous protein production in the methylotrophic yeast Candida Boidinii.
Biotechnology and applied biochemistry, 2009Co-Authors: Hiroya Yurimoto, Yasuyoshi SakaiAbstract:Methylotrophic yeasts can obtain all of their required carbon and energy from methanol, a promising feedstock for biotechnological and chemical processes and a potential replacement for coal and petroleum. When these yeast cells are cultured in the presence of methanol, the enzymes involved in methanol metabolism are massively produced, indicating that the gene promoters of these enzymes are strong methanol-inducible promoters. Using these promoters, high-level heterologous gene expression systems have been developed in several methylotrophic yeast strains, including Pichia pastoris, Hansenula polymorpha, Pichia methanolica and Candida Boidinii. In the present minireview we describe recent insights into the molecular basis of methanol-inducible gene expression in C. Boidinii. In addition, the utility of C. Boidinii gene expression systems for the production of various enzymes is also reviewed.
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Gene-tagging mutagenesis in the methylotrophic yeast Candida Boidinii.
Journal of bioscience and bioengineering, 2007Co-Authors: Yu Sasano, Hiroya Yurimoto, Yasuyoshi SakaiAbstract:A gene-tagging mutagenesis method by random integration of linear DNA fragments was developed and used in the methylotrophic yeast Candida Boidinii to isolate mutants defective in methanol-inducible gene expression. A large number of mutants were obtained, indicating that this method is a powerful tool for random mutagenesis in C. Boidinii.
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Physiological role of the glutathione-dependent formaldehyde dehydrogenase in the methylotrophic yeast Candida Boidinii.
Microbiology, 2002Co-Authors: Bumjun Lee, Hiroya Yurimoto, Yasuyoshi Sakai, Nobuo KatoAbstract: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 fld1Δ strain could not grow on methanol as a carbon source under methanol-limited chemostat culture conditions, even with low dilution rates (D
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physiological role of the glutathione dependent formaldehyde dehydrogenase in the methylotrophic yeast Candida Boidinii
Microbiology, 2002Co-Authors: Bumjun Lee, Hiroya Yurimoto, Yasuyoshi Sakai, Nobuo KatoAbstract: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 fld1Δ strain could not grow on methanol as a carbon source under methanol-limited chemostat culture conditions, even with low dilution rates (D<0·05 h−1), 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.
Nobuo Kato - One of the best experts on this subject based on the ideXlab platform.
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Physiological role of the glutathione-dependent formaldehyde dehydrogenase in the methylotrophic yeast Candida Boidinii.
Microbiology, 2002Co-Authors: Bumjun Lee, Hiroya Yurimoto, Yasuyoshi Sakai, Nobuo KatoAbstract: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 fld1Δ strain could not grow on methanol as a carbon source under methanol-limited chemostat culture conditions, even with low dilution rates (D
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physiological role of the glutathione dependent formaldehyde dehydrogenase in the methylotrophic yeast Candida Boidinii
Microbiology, 2002Co-Authors: Bumjun Lee, Hiroya Yurimoto, Yasuyoshi Sakai, Nobuo KatoAbstract: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 fld1Δ strain could not grow on methanol as a carbon source under methanol-limited chemostat culture conditions, even with low dilution rates (D<0·05 h−1), 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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Physiological role of the glutathione-dependent formaldehyde dehydrogenase in the methylotrophic yeast Candida Boidinii.
Microbiology (Reading England), 2002Co-Authors: Bumjun Lee, Hiroya Yurimoto, Yasuyoshi Sakai, Nobuo KatoAbstract: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 fld1Delta strain could not grow on methanol as a carbon source under methanol-limited chemostat culture conditions, even with low dilution rates (D
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Characterization and high-level production of D-amino acid oxidase in Candida Boidinii
Bioscience biotechnology and biochemistry, 2001Co-Authors: Hiroya Yurimoto, Yasuyoshi Sakai, Tetsuya Hasegawa, Nobuo KatoAbstract:D-Amino acid oxidase (DAO, EC 1.4.3.3) from a methylotrophic yeast, Candida Boidinii, was produced at a high level under the control of the alcohol oxidase gene promoter in the original host. The enzyme was a peroxisomal and monomeric enzyme, and contained noncovalently-bound FAD as a cofactor. The enzyme was active toward several D-amino acids such as D-Ala, D-Met, and D-Ser. An alcohol oxidase-depleted strain (aod1Δ) was found to be a more suitable host for DAO production than the wild-type strain. Several post-translational effects may be responsible for the improvement of the DAO productivity by the aod1Δ strain. Finally, an aod1Δ strain transformant having multi-copies of an expression plasmid on its chromosome could produce DAO amounting up to 30% of the total soluble proteins.
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A Methylotrophic Pathway Participates in Pectin Utilization by Candida Boidinii
Applied and environmental microbiology, 2000Co-Authors: Tomoyuki Nakagawa, Hiroya Yurimoto, Yasuyoshi Sakai, Nobuo Kato, Tatsuro Miyaji, Noboru TomizukaAbstract:The methylotrophic yeast Candida Boidinii S2 was found to be able to grow on pectin or polygalacturonate as a carbon source. When cells were grown on 1% (wt/vol) pectin, C. Boidinii exhibited induced levels of the pectin-depolymerizing enzymes pectin methylesterase (208 mU/mg of protein), pectin lyase (673 mU/mg), pectate lyase (673 mU/mg), and polygalacturonase (3.45 U/mg) and two methanol-metabolizing peroxisomal enzymes, alcohol oxidase (0.26 U/mg) and dihydroxyacetone synthase (94 mU/mg). The numbers of peroxisomes also increased ca. two- to threefold in cells grown on these pectic compounds (3.34 and 2.76 peroxisomes/cell for cells grown on pectin and polygalacturonate, respectively) compared to the numbers in cells grown on glucose (1.29 peroxisomes/cell). The cell density obtained with pectin increased as the degree of methyl esterification of pectic compounds increased, and it decreased in strains from which genes encoding alcohol oxidase and dihydroxyacetone synthase were deleted and in a peroxisome assembly mutant. Our study showed that methanol metabolism and peroxisome assembly play important roles in the degradation of pectin, especially in the utilization of its methyl ester moieties.
Hiroya Yurimoto - One of the best experts on this subject based on the ideXlab platform.
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Msn5p is involved in formaldehyde resistance but not in oxidative stress response in the methylotrophic yeast Candida Boidinii.
Bioscience biotechnology and biochemistry, 2012Co-Authors: Zhenyu Zhai, Hiroya Yurimoto, Yasuyoshi SakaiAbstract:Methylotrophic yeasts, which can utilize methanol as sole carbon and energy source, are exposed to two toxic metabolic intermediates, formaldehyde and hydrogen peroxide, during growth on methanol. Here we report that Msn5p, an importin-β family nuclear exporter, participated in the formaldehyde resistance mechanism but not in the hydrogen peroxide resistance mechanism in Candida Boidinii. Disruption of the MSN5 gene in this yeast caused retardation of growth on formaldehyde-generating growth substrates such as methanol and methylamine, but the expression levels of the methanol-metabolizing enzymes did not fall. The Msn5p-depleted strain was sensitive to formaldehyde but not to hydrogen peroxide. Furthermore, a yellow fluorescent protein-tagged Msn5p was diffuse in the cytoplasm of C. Boidinii when the cells were treated with high concentrations of formaldehyde or ethanol, but was predominantly associated with the nuclei following treatment with hydrogen peroxide.
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Methanol-inducible gene expression and heterologous protein production in the methylotrophic yeast Candida Boidinii.
Biotechnology and applied biochemistry, 2009Co-Authors: Hiroya Yurimoto, Yasuyoshi SakaiAbstract:Methylotrophic yeasts can obtain all of their required carbon and energy from methanol, a promising feedstock for biotechnological and chemical processes and a potential replacement for coal and petroleum. When these yeast cells are cultured in the presence of methanol, the enzymes involved in methanol metabolism are massively produced, indicating that the gene promoters of these enzymes are strong methanol-inducible promoters. Using these promoters, high-level heterologous gene expression systems have been developed in several methylotrophic yeast strains, including Pichia pastoris, Hansenula polymorpha, Pichia methanolica and Candida Boidinii. In the present minireview we describe recent insights into the molecular basis of methanol-inducible gene expression in C. Boidinii. In addition, the utility of C. Boidinii gene expression systems for the production of various enzymes is also reviewed.
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Gene-tagging mutagenesis in the methylotrophic yeast Candida Boidinii.
Journal of bioscience and bioengineering, 2007Co-Authors: Yu Sasano, Hiroya Yurimoto, Yasuyoshi SakaiAbstract:A gene-tagging mutagenesis method by random integration of linear DNA fragments was developed and used in the methylotrophic yeast Candida Boidinii to isolate mutants defective in methanol-inducible gene expression. A large number of mutants were obtained, indicating that this method is a powerful tool for random mutagenesis in C. Boidinii.
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Physiological role of the glutathione-dependent formaldehyde dehydrogenase in the methylotrophic yeast Candida Boidinii.
Microbiology, 2002Co-Authors: Bumjun Lee, Hiroya Yurimoto, Yasuyoshi Sakai, Nobuo KatoAbstract: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 fld1Δ strain could not grow on methanol as a carbon source under methanol-limited chemostat culture conditions, even with low dilution rates (D
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physiological role of the glutathione dependent formaldehyde dehydrogenase in the methylotrophic yeast Candida Boidinii
Microbiology, 2002Co-Authors: Bumjun Lee, Hiroya Yurimoto, Yasuyoshi Sakai, Nobuo KatoAbstract: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 fld1Δ strain could not grow on methanol as a carbon source under methanol-limited chemostat culture conditions, even with low dilution rates (D<0·05 h−1), 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.
Joel M. Goodman - One of the best experts on this subject based on the ideXlab platform.
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Monomeric alcohol oxidase is preferentially digested by a novel protease from Candida Boidinii.
Biochimica et biophysica acta, 2002Co-Authors: Mary Q. Stewart, Ralf Van Dijk, Marten Veenhuis, Joel M. GoodmanAbstract:A protease activity has been partially purified from peroxisomal matrix fractions of the methylotrophic yeast Candida Boidinii. The enzyme migrates as a single peak on a sucrose velocity gradient with an apparent native molecular mass of approximately 80-90 kDa. Activity can be recovered from nonreducing sodium dodecyl sulfate gels as a approximately 20 kDa species, suggesting it is an oligomer. The protein exhibits chymotrypsin-like activity and cleaves the model compound suc-L-L-V-Y-AMC. Additionally, monomers of alcohol oxidase (AO), an abundant protein of C. Boidinii peroxisomes, generated in vitro or in pulse-radiolabeled cells, are preferentially sensitive to degradation by the protease. Sensitivity is lost over time in vivo as AO folds and matures into octamers, suggesting that the protease may be involved in these processes.
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Monomeric alcohol oxidase is preferentially digested by a novel protease from Candida Boidinii
Biochimica et Biophysica Acta (BBA) - Molecular Cell Research, 2002Co-Authors: Mary Q. Stewart, Ralf Van Dijk, Marten Veenhuis, Joel M. GoodmanAbstract:AbstractA protease activity has been partially purified from peroxisomal matrix fractions of the methylotrophic yeast Candida Boidinii. The enzyme migrates as a single peak on a sucrose velocity gradient with an apparent native molecular mass of ∼80–90 kDa. Activity can be recovered from nonreducing sodium dodecyl sulfate gels as a ∼20 kDa species, suggesting it is an oligomer. The protein exhibits chymotrypsin-like activity and cleaves the model compound suc-L-L-V-Y-AMC. Additionally, monomers of alcohol oxidase (AO), an abundant protein of C. Boidinii peroxisomes, generated in vitro or in pulse-radiolabeled cells, are preferentially sensitive to degradation by the protease. Sensitivity is lost over time in vivo as AO folds and matures into octamers, suggesting that the protease may be involved in these processes
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The peroxisomal membrane proteins of Candida Boidinii: gene isolation and expression.
Yeast (Chichester England), 1994Co-Authors: Myrthala Moreno, Richard H. Lark, Kimberly L. Campbell, Joel M. GoodmanAbstract:Candida Boidinii is a methylotrophic yeast in which several growth substrates can cause vigorous peroxisomal proliferation. While such diverse substrates as methanol, oleic acid and D-alanine induce different peroxisomal metabolic pathways, membranes seem to contain common abundant peroxisomal membrane proteins (PMPs). These proteins have been termed PMP31, PMP32 and PMP47. The gene encoding PMP47 has been previously cloned and analysed. We now report the isolation of a second PMP47 gene (or allele) as well as PMP31 and PMP32. PMP47A and PMP47B share 95% sequence identity at the amino acid level. PMP31 and PMP32 each contain 256 amino acids and are highly similar (97% identity) in protein sequence. Both PMP31 and PMP32 are predicted to span the membrane once or twice. All abundant PMPs of C. Boidinii are basic in charge; they all have predicted isoelectric points above 10. RNAs corresponding to the PMP47s and to PMPs31-32 are strongly induced by methanol, oleic acid and D-alanine. While the PMP47s probably encode substrate carriers, the functions of PMP31 and PMP32 from C. Boidinii are still unknown.
T. W. Jeffries - One of the best experts on this subject based on the ideXlab platform.
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Xylitol formation by Candida Boidinii in oxygen limited chemostat culture
Biotechnology Letters, 1996Co-Authors: E. Winkelhausen, P. Pittman, S. Kuzmanova, T. W. JeffriesAbstract:Production of xylitol by Candida Boidinii NRRL Y-17213 occurs under conditions of an oxygen limitation. The extent to which substrate is converted to xylitol and its coproducts (ethanol, other polyols, acetic acid), and the relative flow rates of substrate to energetic and biosynthetic pathways is controlled by the degree of oxygen limitation. With decrease in oxygen concentration in the inlet gas, for a constant dilution rate of 0.05 1/h. the specific oxygen uptake rate decreased from 1.30 to 0.36 mmol/gh Xylitol was not produced at specific oxygen uptake rates above 0.91 mmol/gh. Upon shift to lower oxygen rates, specific xylitol production rate increased more rapidly than specific ethanol production rate:
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FED-BATCH CULTURE FOR XYLITOL PRODUCTION BY Candida Boidinii
Process Biochemistry, 1996Co-Authors: Eleonora Vandeska, S. Kuzmanova, Samuel Amartey, T. W. JeffriesAbstract:Abstract Xylitol production by Candida Boidinii NRRL Y-17213 was investigated in fed-batch fermentations with xylose (50, 100 g litre−1) and a mixture of glucose (25 g litre−1) and xylose (25 g litre−1). All fermentations were initially batch processes with high levels of aeration and rapid production of biomass. Faster growth occurred when a mixture of glucose and xylose, instead of xylose, was used as a substrate; glucose was assimilated first and maximal xylitol production was 39·41 g litre−1, compared with 46·5 and 59·3 g litre−1 in the processes with xylose alone. Fed-batch cultures were characterized with higher xylitol yields (0·57–0·68 g g−1) and production rates (0·32–0·46 g litre−1 h−1) compared with a batch process. Ethanol was accumulated in all processes, in smaller quantities, varying from 11 to 21% of the xylitol concentration. The fed-batch process with the highest initial xylose concentration (100 g litre−1) and the lowest level of aeration in the first phase, resulted in the highest yield of xylitol (75% of theoretical) and the highest productivity (approximately 35% higher than the productivities of the other two fed-batch experiments).
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Xylitol formation and key enzyme activities in Candida Boidinii under different oxygen transfer rates
Journal of Fermentation and Bioengineering, 1995Co-Authors: Eleonora Vandeska, S. Kuzmanova, T. W. JeffriesAbstract:Abstract Under oxygen transfer rates (OTR), from 10 to 30 mmol· l −1 h −1 , Candida Boidinii NRRL Y-17213 exhibited both NADH and NADPH linked d -xylose reductase activities with the former being higher. Xylitol dehydrogenase was mainly NAD dependent. Maximum xylitol production was attained at OTR of 14 mmol· l −1 h −1 . Ethanol, glycerol and ribitol were also produced. A correlation between xylitol accumulation, oxygen availability and key enzyme activities was viewed.
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Effects of environmental conditions on production of xylitol by Candida Boidinii
World journal of microbiology & biotechnology, 1995Co-Authors: Eleonora Vandeska, S. Kuzmanova, Samuel Amartey, T. W. JeffriesAbstract:Candida Boidinii NRRL Y-17213 produced more xylitol thanC. magnolia (NRRL Y-4226 and NRRL Y-7621),Debaryomyces hansenii (C-98 M-21, C-56 M-9 and NRRL Y-7425), orPichia (Hansenula) anomala (NRRL Y-366). WithC. Boidinii, highest xylitol productivity was at pH 7 but highest yield was at pH 8, using 5 g urea and 5 g Casamino acids/I. Decreasing the aeration rate decreased xylose consumption and cell growth but increased the xylitol yield. When an initial cell density of 5.1 g/l was used instead of 1.3 g/l, xylitol yield and the specific xylitol production rate doubled. Substrate concentration had the greatest effect on xylitol production; increasing xylose concentration 7.5-fold (to 150 g/l) gave a 71-fold increase in xylitol production (53 g/l) and a 10-fold increase in xylitol/ethanol ratio. The highest xylitol yield (0.47 g/g), corresponding to 52% of the theoretical yield, was obtained with 150 g xylose/l after 14 days. Xylose at 200 g/l inhibited xylitol production.
