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Takashi Endoh
Kyoto University
7Publications
4H-index
81Citations
Publications 7
Newest
#1Reiko Sakaguchi (Kyoto University)H-Index: 15
#2Takashi Endoh (Kyoto University)H-Index: 4
Last.Takashi Morii (Kyoto University)H-Index: 24
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Abstract A fluorescent sensor for the detection of inositol-1,3,4,5-tetrakisphosphate, Ins(1,3,4,5)P 4 , was constructed from a split PH domain and a single circularly permuted GFP. A structure-based design was conducted to transduce a ligand-induced subtle structural perturbation of the split PH domain to an alteration in the population of the protonated and the deprotonated states of the GFP chromophore. Excitation of each distinct absorption band corresponding to the protonated or the deproto...
#1Tamotsu Kanai (Kyoto University)H-Index: 23
#2Takashi Endoh (Kyoto University)H-Index: 4
Last.Tadayuki Imanaka (Rits: Ritsumeikan University)H-Index: 61
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Previously, we have developed a system for cell-free protein synthesis operated at high temperatures using a lysate of thermococcus kodakaraensis. Here, we tested the synthesis of multiple gene products from a single mRNA. The selected target proteins were TK0944 and TK0943 proteins, the alpha-subunit and the szlig-subunit of acetyl-CoA synthetase III (ACS III) forming an intracellular enzyme complex composed of two beta- and two szlig-subunits (alpha2szlig2). To facilitate the detection and pur...
#1Takashi Endoh (Kyoto University)H-Index: 4
#2Tamotsu Kanai (Kyoto University)H-Index: 23
Last.Tadayuki Imanaka (Kyoto University)H-Index: 61
view all 3 authors...
We have previously established a system for cell-free protein synthesis that can be operated at high temperatures using the cell lysate of a hyperthermophilic archaeon, Thermococcus kodakaraensis. To apply this system to practical use in the field of heterologous protein production, the performance of our system in the synthesis of green fluorescent protein (GFP) was examined. As the wild-type GFP is a thermolabile protein, a thermostable GFP derivative (tGFP) was selected as a candidate for pro...
#1Tamotsu Kanai (Kyoto University)H-Index: 23
#2Takashi Endoh (Kyoto University)H-Index: 4
Last.Tadayuki Imanaka (Kyoto University)H-Index: 61
view all 3 authors...
Previously, we have developed a system for cell-free protein synthesis that can be operated at high temperatures using a lysate of Thermococcus kodakaraensis. Here, we tested cell-free synthesis of green fluorescent protein (GFP) using the T. kodakaraensis system. A thermostable GFP derivative (tGFP) was used as a reporter protein. By changing the codon usage of tGFP gene for T. kodakaraensis, production of tGFP was detectable in a temperature range of 50degC to 65degC, with an optimum at 60degC...
#1Takashi Endoh (Kyoto University)H-Index: 4
#2Tamotsu Kanai (Kyoto University)H-Index: 23
Last.Tadayuki Imanaka (Kyoto University)H-Index: 61
view all 3 authors...
We report in this study an improved system for cell-free protein synthesis at high temperatures using the lysate of Thermococcus kodakaraensis. Previous work indicated that cell-free protein synthesis of ChiAΔ4, a derivative of T. kodakaraensis chitinase, was observed within a temperature range of 40–80°C, and the maximum yield of the ChiAΔ4 synthesized was approximately 1.3 μg/ml. To increase productivity of the system, the following approaches were taken. First, the process of lysate preparati...
#1Takashi Endoh (Kyoto University)H-Index: 4
#2Tamotsu Kanai (Kyoto University)H-Index: 23
Last.Tadayuki Imanaka (Kyoto University)H-Index: 61
view all 7 authors...
Abstract Systems for cell-free protein synthesis available today are usually based on the lysates of either Escherichia coli , wheat germ or rabbit reticulocyte, and protein synthesis reactions using these extracts are performed at moderate temperatures (20–40 °C). We report here the development of a novel system for cell-free protein synthesis that can be operated at high temperatures using a lysate of the hyperthermophilic archaeon, Thermococcus kodakaraensis . With the system, cell-free prote...
#1Tamotsu Kanai (Kyoto University)H-Index: 23
#2Takashi Endoh (Kyoto University)H-Index: 4
Last.Tadayuki Imanaka (Kyoto University)H-Index: 61
view all 7 authors...
We report here the development of a novel system for cell-free protein synthesis that can be operated at high temperatures using a lysate of the hyperthermophilic archaeon, Thermococcus kodakaraensis. With the system, cell-free protein synthesis of ChiADelta4, a derivative of T. kodakaraensis chitinase (ChiA), was observed within a temperature range of 40 degC to 80 degC, with an optimum at 65 degC. Corresponding chitinase activity was also detected in the reaction mixtures after cell-free prote...
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