The Experts below are selected from a list of 252 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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Secretory production of tetrameric native full-length streptavidin with thermostability using Streptomyces lividans as a host
Microbial cell factories, 2015Co-Authors: Shuhei Noda, Takuya Matsumoto, Tsutomu Tanaka, Akihiko KondoAbstract:Background Streptavidin is a tetrameric protein derived from Streptomyces Avidinii, and has tight and specific biotin binding affinity. Applications of the streptavidin-biotin system have been widely studied. Streptavidin is generally produced using protein expression in Escherichia coli. In the present study, the secretory production of streptavidin was carried out using Streptomyces lividans as a host.
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Secretory production of tetrameric native full-length streptavidin with thermostability using Streptomyces lividans as a host
Microbial Cell Factories, 2015Co-Authors: Shuhei Noda, Takuya Matsumoto, Tsutomu Tanaka, Akihiko KondoAbstract:Background Streptavidin is a tetrameric protein derived from Streptomyces Avidinii , and has tight and specific biotin binding affinity. Applications of the streptavidin-biotin system have been widely studied. Streptavidin is generally produced using protein expression in Escherichia coli . In the present study, the secretory production of streptavidin was carried out using Streptomyces lividans as a host. Results In this study, we used the gene encoding native full-length streptavidin, whereas the core region is generally used for streptavidin production in E. coli . Tetrameric streptavidin composed of native full-length streptavidin monomers was successfully secreted in the culture supernatant of S. lividans transformants, and had specific biotin binding affinity as strong as streptavidin produced by E. coli . The amount of Sav using S. lividans was about 9 times higher than using E. coli . Surprisingly, streptavidin produced by S. lividans exhibited affinity to biotin after boiling, despite the fact that tetrameric streptavidin is known to lose its biotin binding ability after brief boiling. Conclusion We successfully produced a large amount of tetrameric streptavidin as a secretory-form protein with unique thermotolerance.
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development of novel yeast cell surface display system for homo oligomeric protein by coexpression of native and anchored subunits
Biotechnology Progress, 2006Co-Authors: Hirotaka Furukawa, Takanori Tanino, Hideki Fukuda, Akihiko KondoAbstract:Streptavidin derived from Streptomyces Avidinii was displayed on the cell surface of the yeast Saccharomyces cerevisiae by cell-surface engineering using two types of plasmid for the expression of a native subunit and an anchored subunit fused with the C-terminus of 318 amino acids of Flo1p containing a glycosylphosphatidylinositol anchor attachment signal. The displayed streptavidin had the binding ability for biotinylated compounds. This was confirmed by fluorescence microscopy after the adsorption of yeast cells displaying streptavidin and biotinylated fluorescein isothiocyanate. On the other hand, streptavidin produced by cells harboring only the plasmid for the expression of the anchored subunit showed a very low binding activity for biotinylated compounds. Cells displaying streptavidin may constitute novel whole-cell affinity adsorbents widely used for immunoassay and biosensing. This coexpression method will ensure that proteins, such as homo- and hetero-oligomeric proteins, are displayed on the cell surface in an active form.
Shuhei Noda - One of the best experts on this subject based on the ideXlab platform.
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Secretory production of tetrameric native full-length streptavidin with thermostability using Streptomyces lividans as a host
Microbial cell factories, 2015Co-Authors: Shuhei Noda, Takuya Matsumoto, Tsutomu Tanaka, Akihiko KondoAbstract:Background Streptavidin is a tetrameric protein derived from Streptomyces Avidinii, and has tight and specific biotin binding affinity. Applications of the streptavidin-biotin system have been widely studied. Streptavidin is generally produced using protein expression in Escherichia coli. In the present study, the secretory production of streptavidin was carried out using Streptomyces lividans as a host.
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Secretory production of tetrameric native full-length streptavidin with thermostability using Streptomyces lividans as a host
Microbial Cell Factories, 2015Co-Authors: Shuhei Noda, Takuya Matsumoto, Tsutomu Tanaka, Akihiko KondoAbstract:Background Streptavidin is a tetrameric protein derived from Streptomyces Avidinii , and has tight and specific biotin binding affinity. Applications of the streptavidin-biotin system have been widely studied. Streptavidin is generally produced using protein expression in Escherichia coli . In the present study, the secretory production of streptavidin was carried out using Streptomyces lividans as a host. Results In this study, we used the gene encoding native full-length streptavidin, whereas the core region is generally used for streptavidin production in E. coli . Tetrameric streptavidin composed of native full-length streptavidin monomers was successfully secreted in the culture supernatant of S. lividans transformants, and had specific biotin binding affinity as strong as streptavidin produced by E. coli . The amount of Sav using S. lividans was about 9 times higher than using E. coli . Surprisingly, streptavidin produced by S. lividans exhibited affinity to biotin after boiling, despite the fact that tetrameric streptavidin is known to lose its biotin binding ability after brief boiling. Conclusion We successfully produced a large amount of tetrameric streptavidin as a secretory-form protein with unique thermotolerance.
Takuya Matsumoto - One of the best experts on this subject based on the ideXlab platform.
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Secretory production of tetrameric native full-length streptavidin with thermostability using Streptomyces lividans as a host
Microbial cell factories, 2015Co-Authors: Shuhei Noda, Takuya Matsumoto, Tsutomu Tanaka, Akihiko KondoAbstract:Background Streptavidin is a tetrameric protein derived from Streptomyces Avidinii, and has tight and specific biotin binding affinity. Applications of the streptavidin-biotin system have been widely studied. Streptavidin is generally produced using protein expression in Escherichia coli. In the present study, the secretory production of streptavidin was carried out using Streptomyces lividans as a host.
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Secretory production of tetrameric native full-length streptavidin with thermostability using Streptomyces lividans as a host
Microbial Cell Factories, 2015Co-Authors: Shuhei Noda, Takuya Matsumoto, Tsutomu Tanaka, Akihiko KondoAbstract:Background Streptavidin is a tetrameric protein derived from Streptomyces Avidinii , and has tight and specific biotin binding affinity. Applications of the streptavidin-biotin system have been widely studied. Streptavidin is generally produced using protein expression in Escherichia coli . In the present study, the secretory production of streptavidin was carried out using Streptomyces lividans as a host. Results In this study, we used the gene encoding native full-length streptavidin, whereas the core region is generally used for streptavidin production in E. coli . Tetrameric streptavidin composed of native full-length streptavidin monomers was successfully secreted in the culture supernatant of S. lividans transformants, and had specific biotin binding affinity as strong as streptavidin produced by E. coli . The amount of Sav using S. lividans was about 9 times higher than using E. coli . Surprisingly, streptavidin produced by S. lividans exhibited affinity to biotin after boiling, despite the fact that tetrameric streptavidin is known to lose its biotin binding ability after brief boiling. Conclusion We successfully produced a large amount of tetrameric streptavidin as a secretory-form protein with unique thermotolerance.
Tsutomu Tanaka - One of the best experts on this subject based on the ideXlab platform.
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Secretory production of tetrameric native full-length streptavidin with thermostability using Streptomyces lividans as a host
Microbial cell factories, 2015Co-Authors: Shuhei Noda, Takuya Matsumoto, Tsutomu Tanaka, Akihiko KondoAbstract:Background Streptavidin is a tetrameric protein derived from Streptomyces Avidinii, and has tight and specific biotin binding affinity. Applications of the streptavidin-biotin system have been widely studied. Streptavidin is generally produced using protein expression in Escherichia coli. In the present study, the secretory production of streptavidin was carried out using Streptomyces lividans as a host.
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Secretory production of tetrameric native full-length streptavidin with thermostability using Streptomyces lividans as a host
Microbial Cell Factories, 2015Co-Authors: Shuhei Noda, Takuya Matsumoto, Tsutomu Tanaka, Akihiko KondoAbstract:Background Streptavidin is a tetrameric protein derived from Streptomyces Avidinii , and has tight and specific biotin binding affinity. Applications of the streptavidin-biotin system have been widely studied. Streptavidin is generally produced using protein expression in Escherichia coli . In the present study, the secretory production of streptavidin was carried out using Streptomyces lividans as a host. Results In this study, we used the gene encoding native full-length streptavidin, whereas the core region is generally used for streptavidin production in E. coli . Tetrameric streptavidin composed of native full-length streptavidin monomers was successfully secreted in the culture supernatant of S. lividans transformants, and had specific biotin binding affinity as strong as streptavidin produced by E. coli . The amount of Sav using S. lividans was about 9 times higher than using E. coli . Surprisingly, streptavidin produced by S. lividans exhibited affinity to biotin after boiling, despite the fact that tetrameric streptavidin is known to lose its biotin binding ability after brief boiling. Conclusion We successfully produced a large amount of tetrameric streptavidin as a secretory-form protein with unique thermotolerance.
Charles R. Cantor - One of the best experts on this subject based on the ideXlab platform.
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An avidin-like domain that does not bind biotin is adopted for oligomerization by the extracellular mosaic protein fibropellin.
Protein science : a publication of the Protein Society, 2005Co-Authors: Itai Yanai, Charles R. Cantor, Xiahui Zhu, Zhiping WengAbstract:The protein avidin found in egg white seems optimized for binding the small vitamin biotin as a stable homotetramer. Indeed, along with its streptavidin ortholog in the bacterium Streptomyces Avidinii, this protein shows the strongest known noncovalent bond of a protein with a small ligand. A third known member of the avidin family, as similar to avidin as is streptavidin, is found at the C-terminal ends of the multidomain fibropellin proteins found in sea urchin. The fibropellins form a layer known as the apical lamina that surrounds the sea urchin embryo throughout development. Based upon the structure of avidin, we deduced a structural model for the avidin-like domain of the fibropellins and found that computational modeling predicts a lack of biotin binding and the preservation of tetramerization. To test this prediction we expressed and purified the fibropellin avidin-like domain and found it indeed to be a homotetramer incapable of binding biotin. Several lines of evidence suggest that the avidin-like domain causes the entire fibropellin protein to tetramerize. We suggest that the presence of the avidin-like domain serves a structural (tetrameric form) rather than functional (biotin-binding) role and may therefore be a molecular instance of exaptation-the modification of an existing function toward a new function. Finally, based upon the oligomerization of the avidin-like domain, we propose a model for the overall structure of the apical lamina.
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[19] Streptavidin-containing chimeric proteins: design and production
Methods in enzymology, 2000Co-Authors: Takeshi Sano, Charles R. CantorAbstract:Publisher Summary This chapter presents the design and production of streptavidin-containing chimeric proteins. It also describes the general protocols for expressing streptavidin-containing chimeras in E. coli by using the bacteriophage T7 expression system and procedures for purifying and characterizing expressed streptavidin-containing chimeric proteins. Streptavidin is a tetrameric protein produced by the bacterium Streptomyces Avidinii , and it has an estimated biotin-binding affinity ( K d ) at 10 –14 M , similar to that of avidin. Its great similarity to avidin, including the biotin-binding and structural characteristics, resulted in the naming of this protein as the Streptomyces equivalent of avidin. The tetrameric structure is essential for its extremely tight biotin-binding affinity because intersubunit contacts to biotin, made by an adjacent subunit through a subunit-subunit interface, have a significant contribution to the biotin-binding site. The three-dimensional structured streptavidin suggests that, without significant modifications, streptavidin would not be able to form a stable, functional molecule in a dimeric or monomeric form, although a dimeric streptavidin with reduced biotin-binding affinity and stability has already been produced. When a partner protein is fused to streptavidin, the resulting streptavidin-containing chimeric protein forms a tetramer via its streptavidin moiety, making the fused partner protein also tetrameric.
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Genetic engineering of streptavidin, a versatile affinity tag
Journal of chromatography. B Biomedical sciences and applications, 1998Co-Authors: Takeshi Sano, Sandor Vajda, Charles R. CantorAbstract:Streptavidin, a tetrameric protein produced by Streptomyces Avidinii, has been used as a useful, versatile affinity tag in a variety of biological applications. The efficacy of streptavidin is derived from its extremely high binding affinity for the vitamin biotin. For the last several years, we have used genetic engineering as a primary means to enhance the properties of streptavidin and to expand the application of streptavidin as an affinity tag. In this review, we describe several genetically engineered streptavidin variants, which include a streptavidin with a reduced biotin-binding affinity, a dimeric streptavidin, and a fusion protein between streptavidin and protein A, along with their potential applications in biological science.
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Streptavidin-based conditional lethal system for biological containment of Pseudomonas putida
Studies in Environmental Science, 1997Co-Authors: Charles R. Cantor, Charlene M. Mello, Przemyslaw Szafranski, Cassandra L Smith, Takeshi Sano, David L KaplanAbstract:The soil bacterium Pseudomonas putida can potentially be applied in bioremediation of areas polluted with aromatic hydrocarbon-based organic solvents and petroleum. However, its use in the open environment has been hindered so far by the lack of reliable survival-control functions. To increase the predictability of P. putida, a novel cell suicide system was constructed, based on triple control of the expression of the lethal Streptomyces Avidinii streptavidin gene (stv) coupled with P. putida TOL plasmid-derived Pm/xylS regulatory circuit. In the absence of hydrocarbon pollutants (3-methylbenzoate in described in vitro studies), all but one in 107 to 108 of genetically modified bacteria commits suicide. The stv-based conditional lethal design can thus effectively limit the spread of released microorganisms strictly to polluted localities and keep them alive only as long as the amount of contaminants keeps above a level determined by the sensitivity of their interaction with the XylS protein.
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A new approach for containment of microorganisms: Dual control of streptavidin expression by antisense RNA and the T7 transcription system
Proceedings of the National Academy of Sciences of the United States of America, 1997Co-Authors: Przemyslaw Szafranski, Charlene M. Mello, Cassandra L Smith, Takeshi Sano, David L Kaplan, Charles R. CantorAbstract:The use of microorganisms in the open environment would be of less concern if they were endowed with programmed self-destruction mechanisms. Here, we propose a new genetic design to increase the effectiveness of cell suicide systems. It ensures very tight control of the derepression of cell death by the combination of the bacteriophage T7 RNA polymerase-lysozyme system and an inducible synthesis of antisense RNA and the Escherichia coli LacI repressor. Functionality of this regulatory concept was tested by applying it to containment of Gram-negative bacteria, based on the conditional expression of the lethal Streptomyces Avidinii streptavidin gene. Toxicity of streptavidin is derived from its exceptionally high binding affinity for an essential prosthetic group, d-biotin. The entire construct was designed to allow the soil bacterium Pseudomonas putida to survive only in the presence of aromatic hydrocarbons and their derivatives which it can degrade. Under favorable growth conditions, clones escaping killing appeared at frequencies of only 10−7–10−8 per cell per generation. The general requirement for biotin through the living world should make streptavidin-based conditional lethal designs applicable to a broad range of containment strategies.