Thermostable ATP Regeneration System Using Polyphosphate Kinase from Thermosynechococcus elongatus BP-1 for D-Amino Acid Dipeptide Synthesis(BIOCHEMICAL ENGINEERING)
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概要
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D-Alanine-D-alanine ligase from Thermotoga maritima ATCC 43589(TmDdl) was a useful biocatalyst for synthesizing D-amino acid dipeptides. TmDdl showed a broad substrate specificity at a high temperature; however, ATP was required for its reaction. One of the methods for an effective ATP supply was the coupling reaction with an ATP regeneration system. However, ATP regeneration systems consisted of enzymes from mesophiles and were difficult to operate at high temperatures. Therefore, an ATP regeneration system that could be used at high temperatures was desired to utilize TmDdl for the effective production of D-amino acid dipeptides. To establish a thermostable ATP regeneration system, polyphosphate kinase from a thermophile, Thermosynechococcus elongatm BP-1(TePpk), was characterized. TePpk showed thermostability up to 70℃; therefore, it was considered that a thermostable ATP regeneration system could be established using TePpk. In the coupling reaction with purified TmDdl and TePpk at 60℃, the amount of ATP required for D-alanyl-D-alanine synthesis could be reduced to 1% of the theoretical amount required when there was no ATP regeneration. When the coupling reaction was applied to a resting cell reaction, ATP was regenerated from an adenosine scaffold in the cell, and D-alanyl-D-alanine was successfully synthesized in the maximum yield of 80%(mol/mol) without the addition of ATP. Thus, an effective synthesis of D-amino acid dipepitides was achieved using the thermostable ATP regeneration system.
- 社団法人日本生物工学会の論文
- 2007-02-25
著者
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Kino Kuniki
Department of Applied Chemistry, Faculty of Science and Engineering, Waseda University
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KIRIMURA Kohtaro
Department of Applied Chemistry, School of Science and Engineering, Waseda University
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Kino K
Department Of Applied Chemistry School Of Science And Engineering Waseda University
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Kino Kuniki
Department Of Applied Chemistry Faculty Of Science And Engineering Waseda University
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Kino Kuniki
Department Of Applied Chemistry School Of Science And Engineering Waseda University
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Sato Masaru
Department of Oral Pathology, Asahi University School of Dentistry
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Kino Kuniki
Dep. Of Applied Chemistry Fac. Of Sci. And Engineering Waseda Univ. 3-4-1 Ohkubo Shinjuku-ku Tokyo 1
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Kirimura Kohtaro
Department Of Applied Chemistry Faculty Of Science And Engineering Waseda University
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Masuda Yusuke
Department of Applied Chemistry, School of Science and Engineering, Waseda University
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Masuda Yusuke
Department Of Applied Chemistry School Of Science And Engineering Waseda University
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Sato Masaru
Department Of Applied Chemistry School Of Advanced Science And Engineering Faculty Of Science And En
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