An Increased Rate of Cell-free Protein Synthesis by Condensing Wheat-germ Extract with Ultrafiltration Membranes
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概要
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Wheat-germ extract for cell-free protein synthesis was condensed with ultrafiltration membranes of which the molecular cut-off values were l0kDa, 100kDa, and 300kDa. Reaction conditions of the cell-free system were optimized for the condensed extracts, which needed a higher concentration of creatine phosphate than the uncondensed one, probably due to the increased activity of degradation of ATP and GTP. By using the condensed extract and optimized reaction conditions, the rate of protein synthesis was increased 2- to 3-fold compared with using an uncondensed extract, and about 10-fold compared with conventional conditions. Condensation of the extract with the 300-kDa membrane showed the highest productivity, which was about 30μg dihydrofolate reductase protein ml^<-1> h^<-1>. The final amount of synthesized protein was one third of that of a continuous-flow cell-free (CFCF) system reported by Endo et al. [J. Biotechnol., 25, 221-230 (1992)] but the productivity was 5-fold higher than that obtained by the CFCF system.
- 社団法人日本農芸化学会の論文
- 1994-04-23
著者
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Nakano Hideo
Laboratory of Molecular Biotechnology, Graduate School of Bioagricultural Sciences, Nagoya Universit
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KAWARASAKI YASUAKI
Laboratory of Molecular Biotechnology, Division of Molecular and Cellular Mechanisms, graduate Schoo
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YAMANE TSUNEO
Laboratory of Molecular Biotechnology, Division of Molecular and Cellular Mechanisms, graduate Schoo
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Yamane Tsuneo
Laboraotory Of Bioreaction Engineering Department Of Food Science And Technology Faculty Of Agricult
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Yamane Tsuneo
Laboratory Of Bioreaction Engineering Department Of Food Science And Technology School Of Agricultur
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Kawarasaki Yasuaki
Laboratory Of Molecular Biotechnology Division Of Molecular And Cellular Mechanisms Graduate School
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Nakano Hideo
Laboratory Of Molecular Biotechnology Graduate School Of Bioagricultural Sciences Nagoya University
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Tanaka Tadaaki
Laboratory of Molecular Biotechnology, Department of Applied Biological Sciences, School of Agricult
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Yamane Tsuneo
Laboratory Of Molecular Biotechnology Graduate School Of Bio- And Agro-sciences Nagoya University
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Tanaka Tadaaki
Laboratory Of Molecular Biotechnology Department Of Applied Biological Sciences School Of Agricultur
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Nakano Hideo
Laboratory Of Molecular Biotechnology Department Of Bioengineering Sciences Graduate School Of Bioag
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