The Experts below are selected from a list of 5610 Experts worldwide ranked by ideXlab platform
Naofumi Kitabatake - One of the best experts on this subject based on the ideXlab platform.
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cloning expression and characterization of recombinant sweet protein thaumatin ii using the Methylotrophic Yeast pichia pastoris
Biotechnology and Bioengineering, 2004Co-Authors: Tetsuya Masuda, Ryosuke Kaneko, Ritsuko Wada, Alka Mehta, Shinobu Tamaki, Yuki Fujita, Naofumi KitabatakeAbstract:: Thaumatin, an intensely sweet-tasting protein, was secreted by the Methylotrophic Yeast Pichia pastoris. The mature thaumatin II gene was directly cloned from Taq polymerase-amplified PCR products by using TA cloning methods and fused the pPIC9K expression vector that contains Saccharomyces cerevisiae prepro alpha-mating factor secretion signal. Several additional amino acid residues were introduced at both the N- and C-terminal ends by genetic modification to investigate the role of the terminal end region for elicitation of sweetness in the thaumatin molecule. The secondary and tertiary structures of purified recombinant thaumatin were almost identical to those of the plant thaumatin molecule. Recombinant thaumatin II elicited a sweet taste as native plant thaumatin II; its threshold value of sweetness to humans was around 50 nM, which is the same as that of plant thaumatin II. These results demonstrate that the functional expression of thaumatin II was attained by Pichia pastoris systems and that the N- and C-terminal regions of the thaumatin II molecule do not -play an important role in eliciting the sweet taste of thaumatin.
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Cloning, expression and characterization of recombinant sweet‐protein thaumatin II using the Methylotrophic Yeast Pichia pastoris
Biotechnology and Bioengineering, 2004Co-Authors: Tetsuya Masuda, Ryosuke Kaneko, Ritsuko Wada, Alka Mehta, Shinobu Tamaki, Yuki Fujita, Naofumi KitabatakeAbstract:: Thaumatin, an intensely sweet-tasting protein, was secreted by the Methylotrophic Yeast Pichia pastoris. The mature thaumatin II gene was directly cloned from Taq polymerase-amplified PCR products by using TA cloning methods and fused the pPIC9K expression vector that contains Saccharomyces cerevisiae prepro alpha-mating factor secretion signal. Several additional amino acid residues were introduced at both the N- and C-terminal ends by genetic modification to investigate the role of the terminal end region for elicitation of sweetness in the thaumatin molecule. The secondary and tertiary structures of purified recombinant thaumatin were almost identical to those of the plant thaumatin molecule. Recombinant thaumatin II elicited a sweet taste as native plant thaumatin II; its threshold value of sweetness to humans was around 50 nM, which is the same as that of plant thaumatin II. These results demonstrate that the functional expression of thaumatin II was attained by Pichia pastoris systems and that the N- and C-terminal regions of the thaumatin II molecule do not -play an important role in eliciting the sweet taste of thaumatin.
Tetsuya Masuda - One of the best experts on this subject based on the ideXlab platform.
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cloning expression and characterization of recombinant sweet protein thaumatin ii using the Methylotrophic Yeast pichia pastoris
Biotechnology and Bioengineering, 2004Co-Authors: Tetsuya Masuda, Ryosuke Kaneko, Ritsuko Wada, Alka Mehta, Shinobu Tamaki, Yuki Fujita, Naofumi KitabatakeAbstract:: Thaumatin, an intensely sweet-tasting protein, was secreted by the Methylotrophic Yeast Pichia pastoris. The mature thaumatin II gene was directly cloned from Taq polymerase-amplified PCR products by using TA cloning methods and fused the pPIC9K expression vector that contains Saccharomyces cerevisiae prepro alpha-mating factor secretion signal. Several additional amino acid residues were introduced at both the N- and C-terminal ends by genetic modification to investigate the role of the terminal end region for elicitation of sweetness in the thaumatin molecule. The secondary and tertiary structures of purified recombinant thaumatin were almost identical to those of the plant thaumatin molecule. Recombinant thaumatin II elicited a sweet taste as native plant thaumatin II; its threshold value of sweetness to humans was around 50 nM, which is the same as that of plant thaumatin II. These results demonstrate that the functional expression of thaumatin II was attained by Pichia pastoris systems and that the N- and C-terminal regions of the thaumatin II molecule do not -play an important role in eliciting the sweet taste of thaumatin.
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Cloning, expression and characterization of recombinant sweet‐protein thaumatin II using the Methylotrophic Yeast Pichia pastoris
Biotechnology and Bioengineering, 2004Co-Authors: Tetsuya Masuda, Ryosuke Kaneko, Ritsuko Wada, Alka Mehta, Shinobu Tamaki, Yuki Fujita, Naofumi KitabatakeAbstract:: Thaumatin, an intensely sweet-tasting protein, was secreted by the Methylotrophic Yeast Pichia pastoris. The mature thaumatin II gene was directly cloned from Taq polymerase-amplified PCR products by using TA cloning methods and fused the pPIC9K expression vector that contains Saccharomyces cerevisiae prepro alpha-mating factor secretion signal. Several additional amino acid residues were introduced at both the N- and C-terminal ends by genetic modification to investigate the role of the terminal end region for elicitation of sweetness in the thaumatin molecule. The secondary and tertiary structures of purified recombinant thaumatin were almost identical to those of the plant thaumatin molecule. Recombinant thaumatin II elicited a sweet taste as native plant thaumatin II; its threshold value of sweetness to humans was around 50 nM, which is the same as that of plant thaumatin II. These results demonstrate that the functional expression of thaumatin II was attained by Pichia pastoris systems and that the N- and C-terminal regions of the thaumatin II molecule do not -play an important role in eliciting the sweet taste of thaumatin.
Yasuyoshi Sakai - One of the best experts on this subject based on the ideXlab platform.
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methanol sensor wsc1 and map kinase suppress degradation of methanol induced peroxisomes in Methylotrophic Yeast
Journal of Cell Science, 2021Co-Authors: Shin Ohsawa, Hiroya Yurimoto, Masahide Oku, Koichi Inoue, Takahiro Isoda, Yasuyoshi SakaiAbstract:In nature, methanol is produced during the hydrolysis of pectin in plant cell walls. Methanol shows circadian dynamics on plant leaves to which methanol-utilizing phyllosphere microorganisms adapt. In the Methylotrophic Yeast Komagataella phaffii (Pichia pastoris), the plasma membrane protein KpWsc1 senses environmental methanol concentrations, and transmits the information to induce genes for methanol metabolism together with huge peroxisomes. In this study, we show that KpWsc1 and its downstream MAPK negatively regulate pexophagy in the presence of >0.15% methanol. Although KpMpk1 was not necessary for expression of methanol-inducible genes and peroxisome biogenesis, KpMpk1, KpRlm1 and a phosphatase were found suppress pexophagy by controlling phosphorylation level of KpAtg30, the key factor of pexophagy. We reveal at the molecular level how the single methanol sensor KpWsc1 commits the cell to peroxisome synthesis and degradation according to the methanol concentration, and discuss the physiological significance of regulating pexophagy for survival in the phyllosphere.
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expression of a codon optimized aspergillus niger pectin methylesterase gene in the Methylotrophic Yeast candida boidinii
Bioscience Biotechnology and Biochemistry, 2014Co-Authors: Kosuke Kawaguchi, Hiroya Yurimoto, Yasuyoshi SakaiAbstract:A codon-optimized Aspergillus niger pectin methylesterase (PME) gene was expressed in the Methylotrophic Yeast Canidia boidinii. The PME-producing strains showed better growth on pectin than the wild-type strains, suggesting that the PME-producing strains could efficiently utilize methyl ester moieties of pectin. On the other hand, overproduction of PME negatively affected the proliferation of C. boidinii on leaves of Arabidopsis thaliana.
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The peroxisomal catalase gene in the Methylotrophic Yeast Pichia methanolica.
Bioscience biotechnology and biochemistry, 2010Co-Authors: Tomoyuki Nakagawa, Hiroya Yurimoto, Yasuyoshi Sakai, Takashi Ito, Shuki Fujimura, Yoshimi Matsufuji, Noboru Tomizuka, Kyoko Yoshida, Akihito Takeuchi, Takashi HayakawaAbstract:In this paper, we describe the CTA1 gene, which encodes a peroxisomal catalase in the Methylotrophic Yeast Pichia methanolica. The P. methanolica CTA1 gene (PmCTA1) comprises a 1,530-bp open reading frame corresponding to a protein of 510 amino acid residues, and its deduced amino acid sequence shows high similarity to those of Cta1ps from other Methylotrophic Yeasts (about 79%). Expression of PmCTA1 in a peroxisomal catalase-depleted (Cbcta1Δ) Candida boidinii strain restored the Methylotrophic growth of the host strain, while the expression of PmCTA1-ΔSRL, which lacks peroxisome targeting signal type 1, did not. In P. methanolica, expression of PmCTA1 was induced when cells were grown on peroxisome-inducing carbon sources, viz., methanol, oleate, and D-alanine. Taken together, these results indicate that PmCTA1 encodes a functional peroxisomal catalase in P. methanolica.
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lag phase autophagy in the Methylotrophic Yeast pichia pastoris
Genes to Cells, 2009Co-Authors: Shun-ichi Yamashita, Hiroya Yurimoto, Dai Murakami, Mari Yoshikawa, Masahide Oku, Yasuyoshi SakaiAbstract:When microbes sense environmental changes, they often temporarily attenuate cell growth to adapt to the new situations, showing a lag phase. In this study, we report that the Methylotrophic Yeast, Pichia pastoris, induced autophagy during the lag phase after the cells were shifted from glucose to methanol medium. Through the autophagic process at least two proteins, aminopeptidase I precursor and cytosolic aldehyde dehydrogenase, were found to be transported into the vacuole, which was dependent on PpAtg11 and PpAtg17, respectively. Notably, PpAtg1 and PpAtg17 were required for early exit from the lag phase during the methanol adaptation. In accordance, phosphorylation states of elongation initiation factor 2α indicated reductions of intracellular amino-acid pools in the atg mutant strains. Together, these data demonstrate the importance of amino acid recycling by autophagy during a cell-remodeling process.
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Lag‐phase autophagy in the Methylotrophic Yeast Pichia pastoris
Genes to cells : devoted to molecular & cellular mechanisms, 2009Co-Authors: Shun-ichi Yamashita, Hiroya Yurimoto, Dai Murakami, Mari Yoshikawa, Masahide Oku, Yasuyoshi SakaiAbstract:When microbes sense environmental changes, they often temporarily attenuate cell growth to adapt to the new situations, showing a lag phase. In this study, we report that the Methylotrophic Yeast, Pichia pastoris, induced autophagy during the lag phase after the cells were shifted from glucose to methanol medium. Through the autophagic process at least two proteins, aminopeptidase I precursor and cytosolic aldehyde dehydrogenase, were found to be transported into the vacuole, which was dependent on PpAtg11 and PpAtg17, respectively. Notably, PpAtg1 and PpAtg17 were required for early exit from the lag phase during the methanol adaptation. In accordance, phosphorylation states of elongation initiation factor 2α indicated reductions of intracellular amino-acid pools in the atg mutant strains. Together, these data demonstrate the importance of amino acid recycling by autophagy during a cell-remodeling process.
Shinobu Tamaki - One of the best experts on this subject based on the ideXlab platform.
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cloning expression and characterization of recombinant sweet protein thaumatin ii using the Methylotrophic Yeast pichia pastoris
Biotechnology and Bioengineering, 2004Co-Authors: Tetsuya Masuda, Ryosuke Kaneko, Ritsuko Wada, Alka Mehta, Shinobu Tamaki, Yuki Fujita, Naofumi KitabatakeAbstract:: Thaumatin, an intensely sweet-tasting protein, was secreted by the Methylotrophic Yeast Pichia pastoris. The mature thaumatin II gene was directly cloned from Taq polymerase-amplified PCR products by using TA cloning methods and fused the pPIC9K expression vector that contains Saccharomyces cerevisiae prepro alpha-mating factor secretion signal. Several additional amino acid residues were introduced at both the N- and C-terminal ends by genetic modification to investigate the role of the terminal end region for elicitation of sweetness in the thaumatin molecule. The secondary and tertiary structures of purified recombinant thaumatin were almost identical to those of the plant thaumatin molecule. Recombinant thaumatin II elicited a sweet taste as native plant thaumatin II; its threshold value of sweetness to humans was around 50 nM, which is the same as that of plant thaumatin II. These results demonstrate that the functional expression of thaumatin II was attained by Pichia pastoris systems and that the N- and C-terminal regions of the thaumatin II molecule do not -play an important role in eliciting the sweet taste of thaumatin.
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Cloning, expression and characterization of recombinant sweet‐protein thaumatin II using the Methylotrophic Yeast Pichia pastoris
Biotechnology and Bioengineering, 2004Co-Authors: Tetsuya Masuda, Ryosuke Kaneko, Ritsuko Wada, Alka Mehta, Shinobu Tamaki, Yuki Fujita, Naofumi KitabatakeAbstract:: Thaumatin, an intensely sweet-tasting protein, was secreted by the Methylotrophic Yeast Pichia pastoris. The mature thaumatin II gene was directly cloned from Taq polymerase-amplified PCR products by using TA cloning methods and fused the pPIC9K expression vector that contains Saccharomyces cerevisiae prepro alpha-mating factor secretion signal. Several additional amino acid residues were introduced at both the N- and C-terminal ends by genetic modification to investigate the role of the terminal end region for elicitation of sweetness in the thaumatin molecule. The secondary and tertiary structures of purified recombinant thaumatin were almost identical to those of the plant thaumatin molecule. Recombinant thaumatin II elicited a sweet taste as native plant thaumatin II; its threshold value of sweetness to humans was around 50 nM, which is the same as that of plant thaumatin II. These results demonstrate that the functional expression of thaumatin II was attained by Pichia pastoris systems and that the N- and C-terminal regions of the thaumatin II molecule do not -play an important role in eliciting the sweet taste of thaumatin.
Alka Mehta - One of the best experts on this subject based on the ideXlab platform.
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cloning expression and characterization of recombinant sweet protein thaumatin ii using the Methylotrophic Yeast pichia pastoris
Biotechnology and Bioengineering, 2004Co-Authors: Tetsuya Masuda, Ryosuke Kaneko, Ritsuko Wada, Alka Mehta, Shinobu Tamaki, Yuki Fujita, Naofumi KitabatakeAbstract:: Thaumatin, an intensely sweet-tasting protein, was secreted by the Methylotrophic Yeast Pichia pastoris. The mature thaumatin II gene was directly cloned from Taq polymerase-amplified PCR products by using TA cloning methods and fused the pPIC9K expression vector that contains Saccharomyces cerevisiae prepro alpha-mating factor secretion signal. Several additional amino acid residues were introduced at both the N- and C-terminal ends by genetic modification to investigate the role of the terminal end region for elicitation of sweetness in the thaumatin molecule. The secondary and tertiary structures of purified recombinant thaumatin were almost identical to those of the plant thaumatin molecule. Recombinant thaumatin II elicited a sweet taste as native plant thaumatin II; its threshold value of sweetness to humans was around 50 nM, which is the same as that of plant thaumatin II. These results demonstrate that the functional expression of thaumatin II was attained by Pichia pastoris systems and that the N- and C-terminal regions of the thaumatin II molecule do not -play an important role in eliciting the sweet taste of thaumatin.
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Cloning, expression and characterization of recombinant sweet‐protein thaumatin II using the Methylotrophic Yeast Pichia pastoris
Biotechnology and Bioengineering, 2004Co-Authors: Tetsuya Masuda, Ryosuke Kaneko, Ritsuko Wada, Alka Mehta, Shinobu Tamaki, Yuki Fujita, Naofumi KitabatakeAbstract:: Thaumatin, an intensely sweet-tasting protein, was secreted by the Methylotrophic Yeast Pichia pastoris. The mature thaumatin II gene was directly cloned from Taq polymerase-amplified PCR products by using TA cloning methods and fused the pPIC9K expression vector that contains Saccharomyces cerevisiae prepro alpha-mating factor secretion signal. Several additional amino acid residues were introduced at both the N- and C-terminal ends by genetic modification to investigate the role of the terminal end region for elicitation of sweetness in the thaumatin molecule. The secondary and tertiary structures of purified recombinant thaumatin were almost identical to those of the plant thaumatin molecule. Recombinant thaumatin II elicited a sweet taste as native plant thaumatin II; its threshold value of sweetness to humans was around 50 nM, which is the same as that of plant thaumatin II. These results demonstrate that the functional expression of thaumatin II was attained by Pichia pastoris systems and that the N- and C-terminal regions of the thaumatin II molecule do not -play an important role in eliciting the sweet taste of thaumatin.