The Experts below are selected from a list of 22212 Experts worldwide ranked by ideXlab platform
Akihiko Kondo - One of the best experts on this subject based on the ideXlab platform.
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fermentation of pigment extracted microalgal residue using yeast Cell Surface Display direct high density ethanol production with competitive life cycle impacts
Green Chemistry, 2020Co-Authors: Xiaochen Huang, Akihiko Kondo, Tomohisa Hasunuma, Shihhsin HoAbstract:Microalgae have attracted increasing attention as a potential feedstock for biofuel production. However, direct high-density ethanol production from microalgae is not commercially feasible due to the requirement for complex pre-treatments and insufficient enzymatic hydrolysis. In this study, we successfully developed a consolidated bioprocessing (CBP) system using recombinant Saccharomyces cerevisiae Displaying synergistic Cellulases/amylases on Cell Surfaces to overcome energy-conversion limitations. As Chlamydomonas sp. JSC4 can accumulate considerable amounts of carbohydrates and pigments (i.e., lutein), performing four rounds of pigment extraction from wet microalgal biomass using acetone was found to significantly eliminate the need for biomass pre-treatment and increase commercial viability. The pigment-extracted JSC4 residues increased ethanol production by 10.7% and 31.6% compared to raw starch and whole JSC4 Cells, respectively. The theoretical ethanol production mass and yield from 300 g L−1 of JSC4 material were 73 g L−1 and 64%, respectively, after fermentation for 72 h in the presence of amylase- and Cellulase-Displaying yeasts, which are dramatically higher than those reported previously. Life cycle assessment (LCA) further revealed that this CBP system has 2.7- to 10.7-fold lower total environmental impact compared to alternative ethanol production methods using microalgal biomass. 2.43 kg ethanol and additional products of 5 g lutein from 1 kg microalgal biomass significantly increased the total economic output to $60.875. Overall, this study successfully demonstrates a feasible Cell-Surface Display fermentation system for use in direct high-density ethanol production from pigment-extracted microalgal material.
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Cell Surface Display technology and metabolic engineering of saccharomyces cerevisiae for enhancing xylitol production from woody biomass
Green Chemistry, 2019Co-Authors: Gregory Guirimand, Chiaki Ogino, Tomohisa Hasunuma, Kentaro Inokuma, Takahiro Bamba, Mami Matsuda, Kenta Morita, Kengo Sasaki, Jeanguy Berrin, Akihiko KondoAbstract:Xylitol is a major commodity chemical widely used in both the food and pharmaceutical industries. Although the worldwide demand for xylitol is constantly growing, its industrial production from purified D-xylose involves a costly and polluting catalytic hydrogenation process. Biotechnological production of xylitol from biomass is a promising strategy to establish an environmentally friendly sustainable conversion process. In this study, xylitol was produced from woody Kraft pulp (KP) by using an engineered strain of Saccharomyces cerevisiae (YPH499-XR-BGL-XYL-XYN) expressing cytosolic xylose reductase (XR), along with β-D-glucosidase (BGL), xylosidase (XYL) and xylanase (XYN) enzymes co-Displayed on the Cell Surface. All these enzymes contributed to the consolidated bioprocessing of KP to xylitol with a yield of 2.3 g L−1 (28% conversion) after 96 hours, along with a significantly reduced amount of commercial enzymes required for pre-treatment (commercial hemiCellulase cocktail (CHC), [CHC] = 0.02 g-DW per g). Further improvement of the Cell Surface Display of XYL and XYN was obtained by using a SED1 “SSS” cassette, containing the coding sequences of the SED1 promoter, the SED1 secretion signal, and the SED1 anchoring domain, to generate the improved strain YPH499-XR-BGL-XYLsss-XYNsss. This improved strain showed a significantly enhanced xylitol production capacity reaching a yield of 3.7 g L−1 (44% conversion) after 96 hours. The Cellulosic content of KP residues was also significantly increased, from 78% to 87% after 96 hours of fermentation, and nanofibrillation of KP residues was observed by scanning electron microscopy. Pre-treatment and fermentation were successfully performed as a proof of concept to further scale up bio-refinery industrial production of xylitol from lignoCellulose.
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recent advances in yeast Cell Surface Display technologies for waste biorefineries
Bioresource Technology, 2016Co-Authors: Shihhsin Ho, Tomohisa Hasunuma, Jo Shu Chang, Akihiko KondoAbstract:Waste biorefinery aims to maximize the output of value-added products from various artificial/agricultural wastes by using integrated bioprocesses. To make waste biorefinery economically feasible, it is thus necessary to develop a low-cost, environment-friendly technique to perform simultaneous biodegradation and bioconversion of waste materials. Cell-Surface Display engineering is a novel, cost-effective technique that can auto-immobilize proteins on the Cell exterior of microorganisms, and has been applied for use with waste biofinery. Through tethering different enzymes (e.g., Cellulase, lipase, and protease) or metal-binding peptides on Cell Surfaces, various yeast strains can effectively produce biofuels and biochemicals from sugar/protein-rich waste materials, catalyze waste oils into biodiesels, or retrieve heavy metals from wastewater. This review critically summarizes recent applications of yeast Cell-Surface Display on various types of waste biorefineries, highlighting its potential and future challenges with regard to commercializing this technology.
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Enhanced Cell-Surface Display and secretory production of Cellulolytic enzymes with Saccharomyces cerevisiae Sed1 signal peptide.
Biotechnology and bioengineering, 2016Co-Authors: Kentaro Inokuma, Tomohisa Hasunuma, Takahiro Bamba, Jun Ishii, Yoichiro Ito, Akihiko KondoAbstract:Recombinant yeast strains Displaying aheterologous Cellulolytic enzymes on their Cell Surfaces using a glycosylphosphatidylinositol (GPI) anchoring system are a promising strategy for bioethanol production from lignoCellulosic materials. A crucial step for Cell wall localization of the enzymes is the intraCellular transport of proteins in yeast Cells. Therefore, the addition of a highly efficient secretion signal sequence is important to increase the amount of the enzymes on the yeast Cell Surface. In this study, we demonstrated the effectiveness of a novel signal peptide (SP) sequence derived from the Saccharomyces cerevisiae SED1 gene for Cell-Surface Display and secretory production of Cellulolytic enzymes. Gene cassettes with SP sequences derived from S. cerevisiae SED1 (SED1SP), Rhizopus oryzae glucoamylase (GLUASP), and S. cerevisiae α-mating pheromone (MFα1SP) were constructed for Cell-Surface Display of Aspergillus aculeatus β-glucosidase (BGL1) and Trichoderma reesei endoglucanase II (EGII). These gene cassettes were integrated into the S. cerevisiae genome. The recombinant strains with the SED1SP showed higher Cell-Surface BGL and EG activities than those with the conventional SP sequences (GLUASP and MFα1SP). The novel SP sequence also improved the secretory production of BGL and EG in S. cerevisiae. The extraCellular BGL activity of the recombinant strains with the SED1SP was 1.3- and 1.9-fold higher than the GLUASP and MFα1SP strains, respectively. Moreover, the utilization of SED1SP successfully enhanced the secretory production of BGL in Pichia pastoris. The utilization of the novel SP sequence is a promising option for highly efficient Cell-Surface Display and secretory production of heterologous proteins in various yeast species. Biotechnol. Bioeng. 2016;113: 2358-2366. © 2016 Wiley Periodicals, Inc.
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Cell Surface engineering of industrial microorganisms for biorefining applications
Biotechnology Advances, 2015Co-Authors: Tsutomu Tanaka, Akihiko KondoAbstract:In order to decrease carbon emissions and negative environmental impacts of various pollutants, biofuel/biochemical production should be promoted for replacing fossil-based industrial processes. Utilization of abundant lignoCellulosic biomass as a feedstock has recently become an attractive option. In this review, we focus on recent efforts of Cell Surface Display using industrial microorganisms such as Escherichia coli and yeast. Cell Surface Display is used primarily for endowing Cellulolytic activity on the host Cells, and enables direct fermentation to generate useful fuels and chemicals from lignoCellulosic biomass. Cell Surface Display systems are systematically summarized, and the drawbacks/perspectives as well as successful application of Surface Display for industrial biotechnology are discussed.
Yuzuru Suzuki - One of the best experts on this subject based on the ideXlab platform.
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expression and presentation of immune related membrane proteins of fish by a Cell Surface Display platform using insect Cells
Molecular Immunology, 2019Co-Authors: Satoshi Tasumi, Shihori Takanashi, Shuichi Asakawa, Kiyoshi Kikuchi, Osamu Nakamura, Keisuke Kobayashi, Yuzuru SuzukiAbstract:Abstract Cell Surface Display is a useful platform to examine the interactions between two proteins of interest, such as immune receptors and ligands. This technique is also useful for studies on the immune receptors of lower vertebrates and invertebrates. However, in many cases, the commonly used Cell culture temperature is relatively high for proteins from such organisms. Since insect Cells can be cultured at lower temperatures than many other Cells, and since they are equipped with “quality control” system, which is advantageous for the presentation of properly folded proteins, we anticipated that the insect Cell Surface Display system could be more suitable for that type of research. In the present study, multiple cloning site of the commercially available expression vector pIB/V5-His was modified, and whether this vector could be useful to present fish immune-related membrane proteins was investigated. Using this plasmid, fugu’s CD8α and CC chemokine receptor 7 could be presented on the Cell Surface. The clones of the lamprey variable lymphocyte receptors obtained previously by the yeast Surface Display (YSD) system as hen’s egg lysozyme (HEL) binders also could be presented on the Cell Surface and bound to HEL. These results suggest that functional immune-related membrane proteins can be presented on the insect Cell Surface, indicating that this system is useful for immunological studies on exothermal animals.
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expression and presentation of immune related membrane proteins of fish by a Cell Surface Display platform using insect Cells
Molecular Immunology, 2019Co-Authors: Satoshi Tasumi, Shihori Takanashi, Shuichi Asakawa, Kiyoshi Kikuchi, Osamu Nakamura, Keisuke Kobayashi, Yuzuru SuzukiAbstract:Abstract Cell Surface Display is a useful platform to examine the interactions between two proteins of interest, such as immune receptors and ligands. This technique is also useful for studies on the immune receptors of lower vertebrates and invertebrates. However, in many cases, the commonly used Cell culture temperature is relatively high for proteins from such organisms. Since insect Cells can be cultured at lower temperatures than many other Cells, and since they are equipped with “quality control” system, which is advantageous for the presentation of properly folded proteins, we anticipated that the insect Cell Surface Display system could be more suitable for that type of research. In the present study, multiple cloning site of the commercially available expression vector pIB/V5-His was modified, and whether this vector could be useful to present fish immune-related membrane proteins was investigated. Using this plasmid, fugu’s CD8α and CC chemokine receptor 7 could be presented on the Cell Surface. The clones of the lamprey variable lymphocyte receptors obtained previously by the yeast Surface Display (YSD) system as hen’s egg lysozyme (HEL) binders also could be presented on the Cell Surface and bound to HEL. These results suggest that functional immune-related membrane proteins can be presented on the insect Cell Surface, indicating that this system is useful for immunological studies on exothermal animals.
Tomohisa Hasunuma - One of the best experts on this subject based on the ideXlab platform.
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fermentation of pigment extracted microalgal residue using yeast Cell Surface Display direct high density ethanol production with competitive life cycle impacts
Green Chemistry, 2020Co-Authors: Xiaochen Huang, Akihiko Kondo, Tomohisa Hasunuma, Shihhsin HoAbstract:Microalgae have attracted increasing attention as a potential feedstock for biofuel production. However, direct high-density ethanol production from microalgae is not commercially feasible due to the requirement for complex pre-treatments and insufficient enzymatic hydrolysis. In this study, we successfully developed a consolidated bioprocessing (CBP) system using recombinant Saccharomyces cerevisiae Displaying synergistic Cellulases/amylases on Cell Surfaces to overcome energy-conversion limitations. As Chlamydomonas sp. JSC4 can accumulate considerable amounts of carbohydrates and pigments (i.e., lutein), performing four rounds of pigment extraction from wet microalgal biomass using acetone was found to significantly eliminate the need for biomass pre-treatment and increase commercial viability. The pigment-extracted JSC4 residues increased ethanol production by 10.7% and 31.6% compared to raw starch and whole JSC4 Cells, respectively. The theoretical ethanol production mass and yield from 300 g L−1 of JSC4 material were 73 g L−1 and 64%, respectively, after fermentation for 72 h in the presence of amylase- and Cellulase-Displaying yeasts, which are dramatically higher than those reported previously. Life cycle assessment (LCA) further revealed that this CBP system has 2.7- to 10.7-fold lower total environmental impact compared to alternative ethanol production methods using microalgal biomass. 2.43 kg ethanol and additional products of 5 g lutein from 1 kg microalgal biomass significantly increased the total economic output to $60.875. Overall, this study successfully demonstrates a feasible Cell-Surface Display fermentation system for use in direct high-density ethanol production from pigment-extracted microalgal material.
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Cell Surface Display technology and metabolic engineering of saccharomyces cerevisiae for enhancing xylitol production from woody biomass
Green Chemistry, 2019Co-Authors: Gregory Guirimand, Chiaki Ogino, Tomohisa Hasunuma, Kentaro Inokuma, Takahiro Bamba, Mami Matsuda, Kenta Morita, Kengo Sasaki, Jeanguy Berrin, Akihiko KondoAbstract:Xylitol is a major commodity chemical widely used in both the food and pharmaceutical industries. Although the worldwide demand for xylitol is constantly growing, its industrial production from purified D-xylose involves a costly and polluting catalytic hydrogenation process. Biotechnological production of xylitol from biomass is a promising strategy to establish an environmentally friendly sustainable conversion process. In this study, xylitol was produced from woody Kraft pulp (KP) by using an engineered strain of Saccharomyces cerevisiae (YPH499-XR-BGL-XYL-XYN) expressing cytosolic xylose reductase (XR), along with β-D-glucosidase (BGL), xylosidase (XYL) and xylanase (XYN) enzymes co-Displayed on the Cell Surface. All these enzymes contributed to the consolidated bioprocessing of KP to xylitol with a yield of 2.3 g L−1 (28% conversion) after 96 hours, along with a significantly reduced amount of commercial enzymes required for pre-treatment (commercial hemiCellulase cocktail (CHC), [CHC] = 0.02 g-DW per g). Further improvement of the Cell Surface Display of XYL and XYN was obtained by using a SED1 “SSS” cassette, containing the coding sequences of the SED1 promoter, the SED1 secretion signal, and the SED1 anchoring domain, to generate the improved strain YPH499-XR-BGL-XYLsss-XYNsss. This improved strain showed a significantly enhanced xylitol production capacity reaching a yield of 3.7 g L−1 (44% conversion) after 96 hours. The Cellulosic content of KP residues was also significantly increased, from 78% to 87% after 96 hours of fermentation, and nanofibrillation of KP residues was observed by scanning electron microscopy. Pre-treatment and fermentation were successfully performed as a proof of concept to further scale up bio-refinery industrial production of xylitol from lignoCellulose.
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recent advances in yeast Cell Surface Display technologies for waste biorefineries
Bioresource Technology, 2016Co-Authors: Shihhsin Ho, Tomohisa Hasunuma, Jo Shu Chang, Akihiko KondoAbstract:Waste biorefinery aims to maximize the output of value-added products from various artificial/agricultural wastes by using integrated bioprocesses. To make waste biorefinery economically feasible, it is thus necessary to develop a low-cost, environment-friendly technique to perform simultaneous biodegradation and bioconversion of waste materials. Cell-Surface Display engineering is a novel, cost-effective technique that can auto-immobilize proteins on the Cell exterior of microorganisms, and has been applied for use with waste biofinery. Through tethering different enzymes (e.g., Cellulase, lipase, and protease) or metal-binding peptides on Cell Surfaces, various yeast strains can effectively produce biofuels and biochemicals from sugar/protein-rich waste materials, catalyze waste oils into biodiesels, or retrieve heavy metals from wastewater. This review critically summarizes recent applications of yeast Cell-Surface Display on various types of waste biorefineries, highlighting its potential and future challenges with regard to commercializing this technology.
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Enhanced Cell-Surface Display and secretory production of Cellulolytic enzymes with Saccharomyces cerevisiae Sed1 signal peptide.
Biotechnology and bioengineering, 2016Co-Authors: Kentaro Inokuma, Tomohisa Hasunuma, Takahiro Bamba, Jun Ishii, Yoichiro Ito, Akihiko KondoAbstract:Recombinant yeast strains Displaying aheterologous Cellulolytic enzymes on their Cell Surfaces using a glycosylphosphatidylinositol (GPI) anchoring system are a promising strategy for bioethanol production from lignoCellulosic materials. A crucial step for Cell wall localization of the enzymes is the intraCellular transport of proteins in yeast Cells. Therefore, the addition of a highly efficient secretion signal sequence is important to increase the amount of the enzymes on the yeast Cell Surface. In this study, we demonstrated the effectiveness of a novel signal peptide (SP) sequence derived from the Saccharomyces cerevisiae SED1 gene for Cell-Surface Display and secretory production of Cellulolytic enzymes. Gene cassettes with SP sequences derived from S. cerevisiae SED1 (SED1SP), Rhizopus oryzae glucoamylase (GLUASP), and S. cerevisiae α-mating pheromone (MFα1SP) were constructed for Cell-Surface Display of Aspergillus aculeatus β-glucosidase (BGL1) and Trichoderma reesei endoglucanase II (EGII). These gene cassettes were integrated into the S. cerevisiae genome. The recombinant strains with the SED1SP showed higher Cell-Surface BGL and EG activities than those with the conventional SP sequences (GLUASP and MFα1SP). The novel SP sequence also improved the secretory production of BGL and EG in S. cerevisiae. The extraCellular BGL activity of the recombinant strains with the SED1SP was 1.3- and 1.9-fold higher than the GLUASP and MFα1SP strains, respectively. Moreover, the utilization of SED1SP successfully enhanced the secretory production of BGL in Pichia pastoris. The utilization of the novel SP sequence is a promising option for highly efficient Cell-Surface Display and secretory production of heterologous proteins in various yeast species. Biotechnol. Bioeng. 2016;113: 2358-2366. © 2016 Wiley Periodicals, Inc.
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Combined Cell-Surface Display- And Secretion-Based Strategies for Production of Cellulosic Ethanol With Saccharomyces Cerevisiae
Biotechnology for biofuels, 2015Co-Authors: Zhuo Liu, Tomohisa Hasunuma, Kentaro Inokuma, Riaan Den Haan, Willem H. Van Zyl, Akihiko KondoAbstract:Engineering Saccharomyces cerevisiae to produce heterologous Cellulases is considered as a promising strategy for production of bioethanol from lignoCellulose. The production of Cellulase is usually pursued by one of the two strategies: Displaying enzyme on the Cell Surface or secreting enzyme into the medium. However, to our knowledge, the combination of the two strategies in a yeast strain has not been employed. In this study, heterologous endoglucanase (EG) and Cellobiohydrolase I (CBHI) were produced in a β-glucosidase Displaying S. cerevisiae strain using Cell-Surface Display, secretion, or a combined strategy. Strains EG-D-CBHI-D and EG-S-CBHI-S (with both enzymes Displayed on the Cell Surface or with both enzymes secreted to the surrounding medium) showed higher ethanol production (2.9 and 2.6 g/L from 10 g/L phosphoric acid swollen Cellulose, respectively), than strains EG-D-CBHI-S and EG-S-CBHI-D (with EG Displayed on Cell Surface and CBHI secreted, or vice versa). After 3-cycle repeated-batch fermentation, the Cellulose degradation ability of strain EG-D-CBHI-D remained 60 % of the 1st batch, at a level that was 1.7-fold higher than that of strain EG-S-CBHI-S. This work demonstrated that placing EG and CBHI in the same space (on the Cell Surface or in the medium) was favorable for amorphous Cellulose-based ethanol fermentation. In addition, the Cellulolytic yeast strain that produced enzymes by the Cell-Surface Display strategy performed better in Cell-recycle batch fermentation compared to strains producing enzymes via the secretion strategy.
Henning D. Mootz - One of the best experts on this subject based on the ideXlab platform.
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bacterial Cell Surface Display of semisynthetic cyclic peptides
ChemBioChem, 2019Co-Authors: Shubhendu Palei, Kira S. Becher, Christian Nienberg, Joachim Jose, Henning D. MootzAbstract:Semisynthetic cyclic peptides containing both non-proteinogenic building blocks, as the synthetic part, and a genetically encoded sequence amenable to DNA-based randomization hold great potential to expand the chemical space in the quest for novel bioactive peptides. Key to an efficient selection of novel binders to biomacromolecules is a robust method to link their genotype and phenotype. A novel bacterial Cell Surface Display technology has been developed to present cyclic peptides composed of synthetic and genetically encoded fragments in their backbones. The fragments were combined by protein trans-splicing and intramolecular oxime ligation. To this end, a split intein half and an unnatural amino acid were Displayed with the genetically encoded part on the Surface of Escherichia coli. Addition of the synthetic fragment equipped with the split intein partner and an aminooxy moiety, as well as the application of a pH-shift protocol, resulted in the onSurface formation of the semisynthetic cyclic peptide. This approach will serve for the generation of cyclic peptide libraries suitable for selection by fluorescence-activated Cell sorting, and more generally enables chemical modification of proteins on the bacterial Surface.
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Bacterial Cell‐Surface Display of Semisynthetic Cyclic Peptides
Chembiochem : a European journal of chemical biology, 2018Co-Authors: Shubhendu Palei, Kira S. Becher, Christian Nienberg, Joachim Jose, Henning D. MootzAbstract:Semisynthetic cyclic peptides containing both non-proteinogenic building blocks, as the synthetic part, and a genetically encoded sequence amenable to DNA-based randomization hold great potential to expand the chemical space in the quest for novel bioactive peptides. Key to an efficient selection of novel binders to biomacromolecules is a robust method to link their genotype and phenotype. A novel bacterial Cell Surface Display technology has been developed to present cyclic peptides composed of synthetic and genetically encoded fragments in their backbones. The fragments were combined by protein trans-splicing and intramolecular oxime ligation. To this end, a split intein half and an unnatural amino acid were Displayed with the genetically encoded part on the Surface of Escherichia coli. Addition of the synthetic fragment equipped with the split intein partner and an aminooxy moiety, as well as the application of a pH-shift protocol, resulted in the onSurface formation of the semisynthetic cyclic peptide. This approach will serve for the generation of cyclic peptide libraries suitable for selection by fluorescence-activated Cell sorting, and more generally enables chemical modification of proteins on the bacterial Surface.
Satoshi Tasumi - One of the best experts on this subject based on the ideXlab platform.
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expression and presentation of immune related membrane proteins of fish by a Cell Surface Display platform using insect Cells
Molecular Immunology, 2019Co-Authors: Satoshi Tasumi, Shihori Takanashi, Shuichi Asakawa, Kiyoshi Kikuchi, Osamu Nakamura, Keisuke Kobayashi, Yuzuru SuzukiAbstract:Abstract Cell Surface Display is a useful platform to examine the interactions between two proteins of interest, such as immune receptors and ligands. This technique is also useful for studies on the immune receptors of lower vertebrates and invertebrates. However, in many cases, the commonly used Cell culture temperature is relatively high for proteins from such organisms. Since insect Cells can be cultured at lower temperatures than many other Cells, and since they are equipped with “quality control” system, which is advantageous for the presentation of properly folded proteins, we anticipated that the insect Cell Surface Display system could be more suitable for that type of research. In the present study, multiple cloning site of the commercially available expression vector pIB/V5-His was modified, and whether this vector could be useful to present fish immune-related membrane proteins was investigated. Using this plasmid, fugu’s CD8α and CC chemokine receptor 7 could be presented on the Cell Surface. The clones of the lamprey variable lymphocyte receptors obtained previously by the yeast Surface Display (YSD) system as hen’s egg lysozyme (HEL) binders also could be presented on the Cell Surface and bound to HEL. These results suggest that functional immune-related membrane proteins can be presented on the insect Cell Surface, indicating that this system is useful for immunological studies on exothermal animals.
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expression and presentation of immune related membrane proteins of fish by a Cell Surface Display platform using insect Cells
Molecular Immunology, 2019Co-Authors: Satoshi Tasumi, Shihori Takanashi, Shuichi Asakawa, Kiyoshi Kikuchi, Osamu Nakamura, Keisuke Kobayashi, Yuzuru SuzukiAbstract:Abstract Cell Surface Display is a useful platform to examine the interactions between two proteins of interest, such as immune receptors and ligands. This technique is also useful for studies on the immune receptors of lower vertebrates and invertebrates. However, in many cases, the commonly used Cell culture temperature is relatively high for proteins from such organisms. Since insect Cells can be cultured at lower temperatures than many other Cells, and since they are equipped with “quality control” system, which is advantageous for the presentation of properly folded proteins, we anticipated that the insect Cell Surface Display system could be more suitable for that type of research. In the present study, multiple cloning site of the commercially available expression vector pIB/V5-His was modified, and whether this vector could be useful to present fish immune-related membrane proteins was investigated. Using this plasmid, fugu’s CD8α and CC chemokine receptor 7 could be presented on the Cell Surface. The clones of the lamprey variable lymphocyte receptors obtained previously by the yeast Surface Display (YSD) system as hen’s egg lysozyme (HEL) binders also could be presented on the Cell Surface and bound to HEL. These results suggest that functional immune-related membrane proteins can be presented on the insect Cell Surface, indicating that this system is useful for immunological studies on exothermal animals.