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Tomotake Morita - One of the best experts on this subject based on the ideXlab platform.
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deficiency of biodegradable plastic degrading enzyme production in a gene deletion mutant of phyllosphere yeast Pseudozyma antarctica defective in mannosylerythritol lipid biosynthesis
AMB Express, 2019Co-Authors: Azusa Saika, Hiroko Kitamoto, Hideaki Koike, Takashi Watanabe, Tohru Yarimizu, Tomotake MoritaAbstract:The basidiomycetous yeast Pseudozyma antarctica (currently designated Moesziomyces antarcticus) produces extracellular enzymes and glycolipids, including mannosylerythritol lipids (MELs), which are biosurfactants. Strain GB-4(0) of this species was previously isolated from rice husks and produces biodegradable plastic-degrading enzyme (Pseudozyma antarctica esterase; PaE). In this study, we generated a MEL biosynthesis-deficient strain (∆PaEMT1) by deleting the gene PaEMT1, which is essential to MEL biosynthesis in strain GB-4(0). The resulting ∆PaEMT1 strain showed deficient PaE activity, and the corresponding signal was hardly detected in its culture supernatant through western blotting analysis using rabbit anti-PaE serum. On the other hand, the relative expression of the gene PaCLE1, encoding PaE, was identical between GB-4(0) and ∆PaEMT1 based on quantitative real-time PCR. When strain ∆PaEMT1 was grown in culture media supplemented with various surfactants, i.e., Tween20, BRIJ35 and TritonX-100, and MELs, PaE activity and secretion recovered. We also attempted to detect intracellular PaE using cell-free extract, but observed no signal in the soluble or insoluble fractions of ∆PaEMT1. This result suggested that the PaCLE1 gene was not translated to PaE, or that expressed PaE was degraded immediately in ∆PaEMT1. Based on these results, MEL biosynthesis is an important contributor to PaE production.
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construction of a Pseudozyma antarctica strain without foreign dna sequences self cloning strain for high yield production of a biodegradable plastic degrading enzyme
Bioscience Biotechnology and Biochemistry, 2019Co-Authors: Yuka Sameshimayamashita, Tomotake Morita, Hideaki Koike, Ken Suzuki, Takashi Watanabe, Tohru Yarimizu, Takumi Tanaka, Shun Tsuboi, Hiroko KitamotoAbstract:ABSTRACTThe basidiomycetous yeast Pseudozyma antarctica GB-4(0) esterase (PaE) is a promising candidate for accelerating degradation of used biodegradable plastics (BPs). To increase safety and red...
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biosynthesis of mono acylated mannosylerythritol lipid in an acyltransferase gene disrupted mutant of Pseudozyma tsukubaensis
Applied Microbiology and Biotechnology, 2018Co-Authors: Azusa Saika, Hideaki Koike, Tokuma Fukuoka, Shuhei Yamamoto, Takahide Kishimoto, Yu Utashima, Tomotake MoritaAbstract:The basidiomycetous yeast genus Pseudozyma produce large amounts of mannosylerythritol lipids (MELs), which are biosurfactants. A few Pseudozyma strains produce mono-acylated MEL as a minor compound using excess glucose as the sole carbon source. Mono-acylated MEL shows higher hydrophilicity than di-acylated MEL and has great potential for aqueous applications. Recently, the gene cluster involved in the MEL biosynthesis pathway was identified in yeast. Here, we generated an acyltransferase (PtMAC2) deletion strain of P. tsukubaensis 1E5 with uracil auxotrophy as a selectable marker. A PtURA5-mutant with a frameshift mutation in PtURA5 was generated as a uracil auxotroph of strain 1E5 by ultraviolet irradiation on plate medium containing 5-fluoro-orotic acid (5-FOA). In the mutant, PtMAC2 was replaced with a PtURA5 cassette containing the 5' untranslated region (UTR) (2000 bp) and 3' UTR (2000 bp) of PtMAC2 by homologous recombination, yielding strain ΔPtMAC2. Based on TLC and NMR analysis, we found that ΔPtMAC2 accumulates MEL acylated at the C-2' position of the mannose moiety. These results indicate that PtMAC2p catalyzes acylation at the C-3' position of the mannose of MEL.
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Enhanced production of a diastereomer type of mannosylerythritol lipid-B by the basidiomycetous yeast Pseudozyma tsukubaensis expressing lipase genes from Pseudozyma antarctica
Applied Microbiology and Biotechnology, 2017Co-Authors: Azusa Saika, Hideaki Koike, Shuhei Yamamoto, Takahide Kishimoto, Tomotake MoritaAbstract:Basidiomycetous yeasts in the genus Pseudozyma are known to produce extracellular glycolipids called mannosylerythritol lipids (MELs). Pseudozyma tsukubaensis produces a large amount of MEL-B using olive oil as the sole carbon source (> 70 g/L production). The MEL-B produced by P. tsukubaensis is a diastereomer type of MEL-B, which consists of 4- O -β- d -mannopyranosyl-(2 R ,3 S )-erythritol as a sugar moiety, in contrast to the conventional type of MELs produced by P. antarctica , which contain 4- O -β- d mannopyranosyl-(2 S ,3 R )-erythritol. In this study, we attempted to increase the production of the diastereomer type of MEL-B in P. tsukubaensis 1E5 by introducing the genes encoding two lipases, PaLIPAp ( PaLIPA ) and PaLIPBp ( PaLIPB ) from P. antarctica T-34. Strain 1E5 expressing PaLIPA exhibited higher lipase activity than the strain possessing an empty vector, which was used as a negative control. Strains of 1E5 expressing PaLIPA or PaLIPB showed 1.9- and 1.6-fold higher MEL-B production than the negative control strain, respectively, and oil consumption was also accelerated by the introduction of these lipase genes. MEL-B production was estimated using time course analysis in the recombinant strains. Strain 1E5 expressing PaLIPA produced 37.0 ± 1.2 g/L of MEL-B within 4 days of cultivation, whereas the strain expressing an empty vector produced 22.1 ± 7.5 g/L in this time. Overexpression of PaLIPA increased MEL-B production by P. tsukubaensis strain 1E5 from olive oil as carbon source by more than 1.7-fold.
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Degradation profiles of biodegradable plastic films by biodegradable plastic-degrading enzymes from the yeast Pseudozyma antarctica and the fungus Paraphoma sp. B47-9
Polymer Degradation and Stability, 2017Co-Authors: Shun Sato, Yukiko Shinozaki, Tomotake Morita, Yuka Sameshima-yamashita, Azusa Saika, Ken Suzuki, Tokuma Fukuoka, Hiroshi Habe, Takashi Watanabe, Hiroko KitamotoAbstract:Esterases from the yeast Pseudozyma antarctica (PaE) and the fungus Paraphoma sp. B47-9 (PCLE) can degrade biodegradable plastics (Shinozaki et al., 2013; Suzuki et al., 2014). The degradation profiles of plastic films composed of poly(butylene succinate), poly(butylene succinate-co-adipate), or poly(butylene adipate) by these enzymes were characterized by liquid chromatography-mass spectroscopy in terms of the molecular structures and molecular weights of the degradation products. Monomers and oligomers with molecular weights corresponding to dimers to octamers were identified as products of degradation by PaE in an aqueous reaction solution, irrespective of the type of biodegradable plastic film. Size-exclusion chromatography indicated that the number-average molecular weight of degraded films decreased with reaction time, suggesting that PaE degraded polyester films randomly into monomer units (endo-type degradation). PCLE also degraded polyester films randomly into monomer units, albeit more slowly than did PaE.
Dai Kitamoto - One of the best experts on this subject based on the ideXlab platform.
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draft genome sequence of the yeast Pseudozyma antarctica type strain jcm10317 a producer of the glycolipid biosurfactants mannosylerythritol lipids
Genome Announcements, 2014Co-Authors: Azusa Saika, Hideaki Koike, Tokuma Fukuoka, Dai Kitamoto, Shun Sato, Hiroshi Habe, Tomoyuki Hori, Tomotake MoritaAbstract:ABSTRACT The basidiomycetous yeast Pseudozyma antarctica is known as a producer of industrial enzymes and the extracellular glycolipids, mannosylerythritol lipids. Here, we report the draft genome sequence of the type strain JCM10317. The draft genome assembly has a size of 18.1 Mb and a G+C content of 60.9%, and it consists of 197 scaffolds.
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accumulation of cellobiose lipids under nitrogen limiting conditions by two ustilaginomycetous yeasts Pseudozyma aphidis and Pseudozyma hubeiensis
Fems Yeast Research, 2013Co-Authors: Tomotake Morita, Tokuma Fukuoka, Tomohiro Imura, Dai KitamotoAbstract:Some basidiomycetous yeast strains extracellularly produce cellobiose lipids (CLs), glycolipid biosurfactants which have strong fungicidal activity. The representative CL producer Ustilago maydis produces CLs together with the other glycolipids, mannosylerythritol lipids (MELs); the preference of the two glycolipids is affected considerably by the nitrogen source. To develop new CL producers, 12 MEL producers were cultured under the nitrogen-limited conditions. Pseudozyma aphidis and Pseudozyma. hubeiensis were characterized as new CL producers. CL production was induced on three strains, P. aphidis, Pseudozyma graminicola, and P. hubeiensis under these conditions. The putative homologous genes of U. maydis cyp1, which encodes a P450 monooxygenase, essential for CL biosynthesis, were partially amplified from their genomic DNA. The nucleotide sequences of the gene fragments from P. hubeiensis and P. aphidis shared identities with U. maydis cyp1 of 99% and 78%, respectively. Furthermore, all of the deduced translation products are tightly clustered in the phylogenic tree of the monooxygenase. These results suggest that the genes involved with CL biosynthesis must be widely distributed in the basidiomycetous fungi as well as the MEL biosynthesis genes, and thus, the genus Pseudozyma has great potential as a biosurfactant producer.
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Genome Sequence of the Basidiomycetous Yeast Pseudozyma antarctica T-34, a Producer of the Glycolipid Biosurfactants Mannosylerythritol Lipids.
Genome Announcements, 2013Co-Authors: Tomotake Morita, Emi Ito, Hiroko Hagiwara, Masayuki Machida, Hideaki Koike, Yoshinori Koyama, Tokuma Fukuoka, Tomohiro Imura, Dai KitamotoAbstract:The basidiomycetous yeast Pseudozyma antarctica T-34 is an excellent producer of mannosylerythritol lipids (MELs), members of the multifunctional extracellular glycolipids, from various feedstocks. Here, the genome sequence of P. antarctica T-34 was determined and annotated. Analysis of the sequence might provide insights into the properties of this yeast that make it superior for use in the production of functional glycolipids, leading to the further development of P. antarctica for industrial applications.
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isolation of Pseudozyma churashimaensis sp nov a novel ustilaginomycetous yeast species as a producer of glycolipid biosurfactants mannosylerythritol lipids
Journal of Bioscience and Bioengineering, 2011Co-Authors: Tomotake Morita, Tokuma Fukuoka, Tomohiro Imura, Masako Takashima, Yuki Ogura, Naoto Hirose, Yukishige Kondo, Dai KitamotoAbstract:Abstract An ustilaginomycetous anamorphic yeast species isolated from the leaves of Saccharum officinarum (sugarcane) in Okinawa, Japan, was identified as a novel Pseudozyma species based on morphological and physiological aspects and molecular taxonomic analysis using the D1/D2 domains of the large subunit (26S) rRNA gene and the internal transcribed spacer 1 (ITS1)-5.8S-ITS2 regions. The name Pseudozyma churashimaensis sp. nov. was proposed for the novel species, with JCM 16988 T as the type strain. Interestingly, P. churashimaensis was found to produce glycolipid biosurfactants, a mixture of mannosylerythritol lipids (MELs), including a novel tri-acetylated derivative (MEL-A2), from glucose. The observed critical micelle concentration (CMC) and the surface tension at CMC of MEL-A2 were 1.7 × 10 −6 M and 29.2 mN/m, respectively. Moreover, on a water-penetration scan, MEL-A2 efficiently formed different lyotropic liquid crystalline phases, including the lamella phase at a wide range of concentrations, indicating its excellent surface-active and self-assembling properties. The novel strain of the genus Pseudozyma should thus facilitate the application of glycolipid biosurfactants in combination with other MEL producers.
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yeast extract stimulates production of glycolipid biosurfactants mannosylerythritol lipids by Pseudozyma hubeiensis sy62
Journal of Bioscience and Bioengineering, 2011Co-Authors: Masaaki Konishi, Tomotake Morita, Tokuma Fukuoka, Tomohiro Imura, Dai Kitamoto, Takahiko Nagahama, Yuji HatadaAbstract:We improved the culture conditions for a biosurfactant producing yeast, Pseudozyma hubeiensis SY62. We found that yeast extract greatly stimulates MEL production. Furthermore, we demonstrated a highly efficient production of MELs in the improved medium by fed-batch cultivation. The final concentration of MELs reached 129 ± 8.2 g/l for one week.
Tokuma Fukuoka - One of the best experts on this subject based on the ideXlab platform.
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biosynthesis of mono acylated mannosylerythritol lipid in an acyltransferase gene disrupted mutant of Pseudozyma tsukubaensis
Applied Microbiology and Biotechnology, 2018Co-Authors: Azusa Saika, Hideaki Koike, Tokuma Fukuoka, Shuhei Yamamoto, Takahide Kishimoto, Yu Utashima, Tomotake MoritaAbstract:The basidiomycetous yeast genus Pseudozyma produce large amounts of mannosylerythritol lipids (MELs), which are biosurfactants. A few Pseudozyma strains produce mono-acylated MEL as a minor compound using excess glucose as the sole carbon source. Mono-acylated MEL shows higher hydrophilicity than di-acylated MEL and has great potential for aqueous applications. Recently, the gene cluster involved in the MEL biosynthesis pathway was identified in yeast. Here, we generated an acyltransferase (PtMAC2) deletion strain of P. tsukubaensis 1E5 with uracil auxotrophy as a selectable marker. A PtURA5-mutant with a frameshift mutation in PtURA5 was generated as a uracil auxotroph of strain 1E5 by ultraviolet irradiation on plate medium containing 5-fluoro-orotic acid (5-FOA). In the mutant, PtMAC2 was replaced with a PtURA5 cassette containing the 5' untranslated region (UTR) (2000 bp) and 3' UTR (2000 bp) of PtMAC2 by homologous recombination, yielding strain ΔPtMAC2. Based on TLC and NMR analysis, we found that ΔPtMAC2 accumulates MEL acylated at the C-2' position of the mannose moiety. These results indicate that PtMAC2p catalyzes acylation at the C-3' position of the mannose of MEL.
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Degradation profiles of biodegradable plastic films by biodegradable plastic-degrading enzymes from the yeast Pseudozyma antarctica and the fungus Paraphoma sp. B47-9
Polymer Degradation and Stability, 2017Co-Authors: Shun Sato, Yukiko Shinozaki, Tomotake Morita, Yuka Sameshima-yamashita, Azusa Saika, Ken Suzuki, Tokuma Fukuoka, Hiroshi Habe, Takashi Watanabe, Hiroko KitamotoAbstract:Esterases from the yeast Pseudozyma antarctica (PaE) and the fungus Paraphoma sp. B47-9 (PCLE) can degrade biodegradable plastics (Shinozaki et al., 2013; Suzuki et al., 2014). The degradation profiles of plastic films composed of poly(butylene succinate), poly(butylene succinate-co-adipate), or poly(butylene adipate) by these enzymes were characterized by liquid chromatography-mass spectroscopy in terms of the molecular structures and molecular weights of the degradation products. Monomers and oligomers with molecular weights corresponding to dimers to octamers were identified as products of degradation by PaE in an aqueous reaction solution, irrespective of the type of biodegradable plastic film. Size-exclusion chromatography indicated that the number-average molecular weight of degraded films decreased with reaction time, suggesting that PaE degraded polyester films randomly into monomer units (endo-type degradation). PCLE also degraded polyester films randomly into monomer units, albeit more slowly than did PaE.
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a gene cluster for biosynthesis of mannosylerythritol lipids consisted of 4 o β d mannopyranosyl 2r 3s erythritol as the sugar moiety in a basidiomycetous yeast Pseudozyma tsukubaensis
PLOS ONE, 2016Co-Authors: Azusa Saika, Hideaki Koike, Tokuma Fukuoka, Shuhei Yamamoto, Takahide Kishimoto, Tomotake MoritaAbstract:Mannosylerythritol lipids (MELs) belong to the glycolipid biosurfactants and are produced by various fungi. The basidiomycetous yeast Pseudozyma tsukubaensis produces diastereomer type of MEL-B, which contains 4-O-β-D-mannopyranosyl-(2R,3S)-erythritol (R-form) as the sugar moiety. In this respect it differs from conventional type of MELs, which contain 4-O-β-D-mannopyranosyl-(2S,3R)-erythritol (S-form) as the sugar moiety. While the biosynthetic gene cluster for conventional type of MELs has been previously identified in Ustilago maydis and Pseudozyma antarctica, the genetic basis for MEL biosynthesis in P. tsukubaensis is unknown. Here, we identified a gene cluster involved in MEL biosynthesis in P. tsukubaensis. Among these genes, PtEMT1, which encodes erythritol/mannose transferase, had greater than 69% identity with homologs from strains in the genera Ustilago, Melanopsichium, Sporisorium and Pseudozyma. However, phylogenetic analysis placed PtEMT1p in a separate clade from the other proteins. To investigate the function of PtEMT1, we introduced the gene into a P. antarctica mutant strain, ΔPaEMT1, which lacks MEL biosynthesis ability owing to the deletion of PaEMT1. Using NMR spectroscopy, we identified the biosynthetic product as MEL-A with altered sugar conformation. These results indicate that PtEMT1p catalyzes the sugar conformation of MELs. This is the first report of a gene cluster for the biosynthesis of diastereomer type of MEL.
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draft genome sequence of the yeast Pseudozyma antarctica type strain jcm10317 a producer of the glycolipid biosurfactants mannosylerythritol lipids
Genome Announcements, 2014Co-Authors: Azusa Saika, Hideaki Koike, Tokuma Fukuoka, Dai Kitamoto, Shun Sato, Hiroshi Habe, Tomoyuki Hori, Tomotake MoritaAbstract:ABSTRACT The basidiomycetous yeast Pseudozyma antarctica is known as a producer of industrial enzymes and the extracellular glycolipids, mannosylerythritol lipids. Here, we report the draft genome sequence of the type strain JCM10317. The draft genome assembly has a size of 18.1 Mb and a G+C content of 60.9%, and it consists of 197 scaffolds.
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accumulation of cellobiose lipids under nitrogen limiting conditions by two ustilaginomycetous yeasts Pseudozyma aphidis and Pseudozyma hubeiensis
Fems Yeast Research, 2013Co-Authors: Tomotake Morita, Tokuma Fukuoka, Tomohiro Imura, Dai KitamotoAbstract:Some basidiomycetous yeast strains extracellularly produce cellobiose lipids (CLs), glycolipid biosurfactants which have strong fungicidal activity. The representative CL producer Ustilago maydis produces CLs together with the other glycolipids, mannosylerythritol lipids (MELs); the preference of the two glycolipids is affected considerably by the nitrogen source. To develop new CL producers, 12 MEL producers were cultured under the nitrogen-limited conditions. Pseudozyma aphidis and Pseudozyma. hubeiensis were characterized as new CL producers. CL production was induced on three strains, P. aphidis, Pseudozyma graminicola, and P. hubeiensis under these conditions. The putative homologous genes of U. maydis cyp1, which encodes a P450 monooxygenase, essential for CL biosynthesis, were partially amplified from their genomic DNA. The nucleotide sequences of the gene fragments from P. hubeiensis and P. aphidis shared identities with U. maydis cyp1 of 99% and 78%, respectively. Furthermore, all of the deduced translation products are tightly clustered in the phylogenic tree of the monooxygenase. These results suggest that the genes involved with CL biosynthesis must be widely distributed in the basidiomycetous fungi as well as the MEL biosynthesis genes, and thus, the genus Pseudozyma has great potential as a biosurfactant producer.
Tomohiro Imura - One of the best experts on this subject based on the ideXlab platform.
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accumulation of cellobiose lipids under nitrogen limiting conditions by two ustilaginomycetous yeasts Pseudozyma aphidis and Pseudozyma hubeiensis
Fems Yeast Research, 2013Co-Authors: Tomotake Morita, Tokuma Fukuoka, Tomohiro Imura, Dai KitamotoAbstract:Some basidiomycetous yeast strains extracellularly produce cellobiose lipids (CLs), glycolipid biosurfactants which have strong fungicidal activity. The representative CL producer Ustilago maydis produces CLs together with the other glycolipids, mannosylerythritol lipids (MELs); the preference of the two glycolipids is affected considerably by the nitrogen source. To develop new CL producers, 12 MEL producers were cultured under the nitrogen-limited conditions. Pseudozyma aphidis and Pseudozyma. hubeiensis were characterized as new CL producers. CL production was induced on three strains, P. aphidis, Pseudozyma graminicola, and P. hubeiensis under these conditions. The putative homologous genes of U. maydis cyp1, which encodes a P450 monooxygenase, essential for CL biosynthesis, were partially amplified from their genomic DNA. The nucleotide sequences of the gene fragments from P. hubeiensis and P. aphidis shared identities with U. maydis cyp1 of 99% and 78%, respectively. Furthermore, all of the deduced translation products are tightly clustered in the phylogenic tree of the monooxygenase. These results suggest that the genes involved with CL biosynthesis must be widely distributed in the basidiomycetous fungi as well as the MEL biosynthesis genes, and thus, the genus Pseudozyma has great potential as a biosurfactant producer.
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Genome Sequence of the Basidiomycetous Yeast Pseudozyma antarctica T-34, a Producer of the Glycolipid Biosurfactants Mannosylerythritol Lipids.
Genome Announcements, 2013Co-Authors: Tomotake Morita, Emi Ito, Hiroko Hagiwara, Masayuki Machida, Hideaki Koike, Yoshinori Koyama, Tokuma Fukuoka, Tomohiro Imura, Dai KitamotoAbstract:The basidiomycetous yeast Pseudozyma antarctica T-34 is an excellent producer of mannosylerythritol lipids (MELs), members of the multifunctional extracellular glycolipids, from various feedstocks. Here, the genome sequence of P. antarctica T-34 was determined and annotated. Analysis of the sequence might provide insights into the properties of this yeast that make it superior for use in the production of functional glycolipids, leading to the further development of P. antarctica for industrial applications.
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isolation of Pseudozyma churashimaensis sp nov a novel ustilaginomycetous yeast species as a producer of glycolipid biosurfactants mannosylerythritol lipids
Journal of Bioscience and Bioengineering, 2011Co-Authors: Tomotake Morita, Tokuma Fukuoka, Tomohiro Imura, Masako Takashima, Yuki Ogura, Naoto Hirose, Yukishige Kondo, Dai KitamotoAbstract:Abstract An ustilaginomycetous anamorphic yeast species isolated from the leaves of Saccharum officinarum (sugarcane) in Okinawa, Japan, was identified as a novel Pseudozyma species based on morphological and physiological aspects and molecular taxonomic analysis using the D1/D2 domains of the large subunit (26S) rRNA gene and the internal transcribed spacer 1 (ITS1)-5.8S-ITS2 regions. The name Pseudozyma churashimaensis sp. nov. was proposed for the novel species, with JCM 16988 T as the type strain. Interestingly, P. churashimaensis was found to produce glycolipid biosurfactants, a mixture of mannosylerythritol lipids (MELs), including a novel tri-acetylated derivative (MEL-A2), from glucose. The observed critical micelle concentration (CMC) and the surface tension at CMC of MEL-A2 were 1.7 × 10 −6 M and 29.2 mN/m, respectively. Moreover, on a water-penetration scan, MEL-A2 efficiently formed different lyotropic liquid crystalline phases, including the lamella phase at a wide range of concentrations, indicating its excellent surface-active and self-assembling properties. The novel strain of the genus Pseudozyma should thus facilitate the application of glycolipid biosurfactants in combination with other MEL producers.
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yeast extract stimulates production of glycolipid biosurfactants mannosylerythritol lipids by Pseudozyma hubeiensis sy62
Journal of Bioscience and Bioengineering, 2011Co-Authors: Masaaki Konishi, Tomotake Morita, Tokuma Fukuoka, Tomohiro Imura, Dai Kitamoto, Takahiko Nagahama, Yuji HatadaAbstract:We improved the culture conditions for a biosurfactant producing yeast, Pseudozyma hubeiensis SY62. We found that yeast extract greatly stimulates MEL production. Furthermore, we demonstrated a highly efficient production of MELs in the improved medium by fed-batch cultivation. The final concentration of MELs reached 129 ± 8.2 g/l for one week.
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production of a novel glycolipid biosurfactant mannosylmannitol lipid by Pseudozyma parantarctica and its interfacial properties
Applied Microbiology and Biotechnology, 2009Co-Authors: Tomotake Morita, Masaaki Konishi, Tokuma Fukuoka, Tomohiro Imura, Shuhei Yamamoto, Masaru Kitagawa, Atsushi Sogabe, Dai KitamotoAbstract:The development of a novel glycolipid biosurfactant was undertaken using the high-level producers of mannosylerythritol lipids (MELs) such as Pseudozyma parantarctica, Pseudozyma antarctica, and Pseudozyma rugulosa. Besides the conventional MELs (MEL-A, MEL-B, and MEL-C), these yeasts produced an unknown glycolipid when they were cultivated in a medium containing 4% (w/v) olive oil and 4% (w/w) mannitol as the carbon source. The unknown glycolipid extracted from the culture medium of P. parantarctica JCM 11752T displayed the spot with lower mobility than that of known MELs on TLC and provided mainly two peaks identical to mannose and mannitol on high-performance liquid chromatography after acid hydrolysis. Based on structural analysis by 1H and 13C nuclear magnetic resonance, the novel glycolipid was composed of mannose and mannitol as the hydrophilic sugar moiety and was identified as mannosylmannitol lipid (MML). Of the strains tested, P. parantarctica JCM 11752T gave the best yield of MML (18.2 g/L), which comprised approximately 35% of all glycolipids produced. We further investigated the interfacial properties of the MML, considering the unique hydrophilic structure. The observed critical micelle concentration (CMC) and the surface tension at CMC of the MML were 2.6 × 10−6 M and 24.2 mN/m, respectively. In addition, on a water-penetration scan, the MML efficiently formed not only the lamella phase (Lα) but also the myelins at a wide range of concentrations, indicating its excellent self-assembling properties and high hydrophilicity. The present glycolipid should thus facilitate the application of biosurfactants as new functional materials.
Teun Boekhout - One of the best experts on this subject based on the ideXlab platform.
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multigene phylogeny and taxonomic revision of yeasts and related fungi in the ustilaginomycotina
Studies in Mycology, 2015Co-Authors: Q M Wang, Teun Boekhout, Fengyan Bai, Dominik Begerow, Marizeth Groenewald, Xinzhan Liu, Bart TheelenAbstract:The subphylum Ustilaginomycotina (Basidiomycota, Fungi) comprises mainly plant pathogenic fungi (smuts). Some of the lineages possess cultivable unicellular stages that are usually classified as yeast or yeast-like species in a largely artificial taxonomic system which is independent from and largely incompatible with that of the smut fungi. Here we performed phylogenetic analyses based on seven genes including three nuclear ribosomal RNA genes and four protein coding genes to address the molecular phylogeny of the ustilaginomycetous yeast species and their filamentous counterparts. Taxonomic revisions were proposed to reflect this phylogeny and to implement the ‘One Fungus = One Name’ principle. The results confirmed that the yeast-containing classes Malasseziomycetes, Moniliellomycetes and Ustilaginomycetes are monophyletic, whereas Exobasidiomycetes in the current sense remains paraphyletic. Four new genera, namely Dirkmeia gen. nov., Kalmanozyma gen. nov., Golubevia gen. nov. and Robbauera gen. nov. are proposed to accommodate Pseudozyma and Tilletiopsis species that are distinct from the other smut taxa and belong to clades that are separate from those containing type species of the hitherto described genera. Accordingly, new orders Golubeviales ord. nov. with Golubeviaceae fam. nov. and Robbauerales ord. nov. with Robbaueraceae fam. nov. are proposed to accommodate the sisterhood of Golubevia gen. nov. and Robbauera gen. nov. with other orders of Exobasidiomycetes. The majority of the remaining anamorphic yeast species are transferred to corresponding teleomorphic genera based on strongly supported phylogenetic affinities, resulting in the proposal of 28 new combinations. The taxonomic status of a few Pseudozyma species remains to be determined because of their uncertain phylogenetic positions. We propose to use the term pro tempore or pro tem. in abbreviation to indicate the single-species lineages that are temporarily maintained.
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Pseudozyma bandoni emend boekhout and a comparison with the yeast state of ustilago maydis de candolle corda
The Yeasts a Taxonomic Study 5th ed, 2011Co-Authors: Teun BoekhoutAbstract:Publisher Summary This chapter focuses on Pseudozyma genus and its member species. The hyphae in this genus occur usually with retraction septa, and the cytoplasm in the cells is separated by lysed cells. Fusiform blastoconidia occur on sterigma-like denticles and may form an aerial mycelium made up of branched, acropetal chains of conidia. Chlamydospores may be present, but ballistoconidia are absent. The colonies are whitish, pinkish, orange, or brownish-yellow, usually dimorphic with the central part yeastlike, and the margin fringed with hyphae. A thin aerial mycelium is usually present, which toward the margin may become zonate. The member species of this genus are Pseudozyma antarctica, Pseudozyma aphidis, Pseudozyma flocculosa, Pseudozyma fusiformata, Pseudozyma prolifica, Pseudozyma rugulosa, Pseudozyma tsukubaensis, and Ustilago maydis. After undergoing growth on 5% malt extract agar for five days at 17° C, the cells of Pseudozyma antarctica, appear as cylindrical to fusiform, varies in size, with 1–3 oil droplets, and as single. The colonies of this species are dimorphic, withthe center flat to somewhat raised, glabrous to velvety hirsute, smooth to somewhat irregularly furrowed, dull, pale creamish-white to yellowish-brown. The cells of Pseudozyma aphidis, after undergoing growth on 5% malt extract agar for five days at 17° C, appear as fusiform, variable in size, and contain many oil droplets.
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molecular and physiological analysis of the powdery mildew antagonist Pseudozyma flocculosa and related fungi
Phytopathology, 2001Co-Authors: Tyler J Avis, Teun Boekhout, S J Caron, Richard C Hamelin, Richard R BelangerAbstract:ABSTRACT A number of phenotypic and genotypic characteristics were used to ascertain the identity and diversity of Pseudozyma flocculosa, a natural antagonist of powdery mildews that has received little attention in terms of taxonomy. To this end, several putative isolates of P. flocculosa as well as several closely related species were analyzed. Ribosomal DNA sequences distinguished P. flocculosa from other Pseudozyma spp. and identified two previously unknown Pseudozyma isolates as P. flocculosa. Random amplified microsatellites revealed three distinct P. flocculosa strains among the tested isolates. Biocontrol properties and antifungal metabolite production were limited to the P. flocculosa spp. Results produced useful molecular markers to (i) distinguish P. flocculosa from other related fungi, (ii) identify different strains within this species, and (iii) aid in the construction of isolate-specific molecular tools that will assist in research and development of P. flocculosa as a biocontrol agent of p...
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phylogenetic placements of ustilaginomycetous anamorphs as deduced from nuclear lsu rdna sequences
Fungal Biology, 2000Co-Authors: Dominik Begerow, Robert Bauer, Teun BoekhoutAbstract:In order to integrate ustilaginomycetous anamorphs into the general phylogenetic system of Ustilaginomycetes, partial nuclear large subunit ribosomal DNA sequences of 56 teleomorphic and 19 anamorphic species of the Ustilaginomycetes were analysed. Maximum parsimony and neighbour joining confirm the new suprageneric system of Ustilaginomycetes and indicate that (i) the species of Pseudozyma represent anamorphs of Ustilaginales parasitizing grasses, (ii) Pseudozyma prolifica, the type of Pseudozyma, is very closely related to Ustilago maydis, (iii) Pseudozyma tsukubaensis is probably synonymous with Ustilago spermophora, (iv) the species of Malassezia represent a group of its own within the Exobasidiomycetidae, (v) Tilletiopsis cremea, T. lilacina and T. washingtonensis belong to the Entylomatales and (vi) T. flava, T. fulvescens and T. minor are members of the Georgefischeriales. Like all Tilletiopsis species tested, T. albescens and T. pallescens are members of the Exobasidiomycetidae, but they cannot be ascribed to any of the known orders of this subclass. The description of the Malasseziales is emended.
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103 Pseudozyma bandoni emend boekhout and a comparison with the yeast state of ustilago maydis de candolle corda
The Yeasts (Fourth Edition)#R##N#A Taxonomic Study, 1998Co-Authors: Teun Boekhout, J W FellAbstract:Publisher Summary This chapter focuses on Pseudozyma genus and its member species. The hyphae in this genus occur usually with retraction septa, and the cytoplasm in the cells is separated by lysed cells. Fusiform blastoconidia occur on sterigma-like denticles and may form an aerial mycelium made up of branched, acropetal chains of conidia. Chlamydospores may be present, but ballistoconidia are absent. The colonies are whitish, pinkish, orange, or brownish-yellow, usually dimorphic with the central part yeastlike, and the margin fringed with hyphae. A thin aerial mycelium is usually present, which toward the margin may become zonate. The member species of this genus are Pseudozyma antarctica, Pseudozyma aphidis, Pseudozyma flocculosa, Pseudozyma fusiformata, Pseudozyma prolifica, Pseudozyma rugulosa, Pseudozyma tsukubaensis, and Ustilago maydis. After undergoing growth on 5% malt extract agar for five days at 17° C, the cells of Pseudozyma antarctica, appear as cylindrical to fusiform, varies in size, with 1–3 oil droplets, and as single. The colonies of this species are dimorphic, withthe center flat to somewhat raised, glabrous to velvety hirsute, smooth to somewhat irregularly furrowed, dull, pale creamish-white to yellowish-brown. The cells of Pseudozyma aphidis, after undergoing growth on 5% malt extract agar for five days at 17° C, appear as fusiform, variable in size, and contain many oil droplets.
