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논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 저널정보
- 한국미생물생명공학회 Journal of Microbiology and Biotechnology Journal of Microbiology and Biotechnology 제25권 제12호
- 발행연도
- 2015.1
- 수록면
- 2,034 - 2,042 (9page)
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초록· 키워드
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A one-pot process of enzymatic synthesis of deoxythymidine-5’-triphosphate (5’-dTTP) employing whole cells of recombinant Escherichia coli coexpressing thymidylate kinase (TMKase) and acetate kinase (ACKase) was developed. Genes tmk and ack from E. coli were cloned and inserted into pET28a(+), and then transduced into E. coli BL21 (DE3) to form recombinant strain pTA in which TMKase and ACKase were simultaneously overexpressed. It was found that the relative residual specific activities of TMKase and ACKase, in pTA pretreated with 20 mM ethylene diamine tetraacetic acid (EDTA) at 25oC for 30 min, were 94% and 96%, respectively. The yield of 5’-dTTP reached above 94% from 5 mM deoxythymidine 5’-monophosphate (5’-dTMP) and 15 mM acetyl phosphate catalyzed with intact cells of pTA pretreated with EDTA. The process was so effective that only 0.125 mM adenosine-5’- triphosphate was sufficient to deliver the phosphate group from acetyl phosphate to dTMP and dTDP.
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참고문헌 (23)
참고문헌 신청
Banerjee G / 2010 / Improving enzymes for biomass conversion: a basic research perspective / Bioenerg. Res 3 : 82 ~ 92
Bradford MM / 1976 / A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein?dye binding / Anal. Biochem 72 :
Canovas M / 2005 / Permeabilization of Escherichia coli cells in the biotransformation of trimethylammonium compounds into L-carnitine / Enzyme Microb. Technol 37 : 300 ~ 308
Chambers RW / 1957 / Nucleoside polyphosphates. V.1 Syntheses of guanosine 5’-di- and triphosphates / J. Am. Chem. Soc 79 : 3752 ~ 3755
Chen RR / 2007 / Permeability issues in whole-cell bioprocesses and cellular membrane engineering / Appl. Microbiol. Biotechnol 74 : 730 ~ 738
Bradford MM / 1976 / A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein?dye binding / Anal. Biochem 72 :
Canovas M / 2005 / Permeabilization of Escherichia coli cells in the biotransformation of trimethylammonium compounds into L-carnitine / Enzyme Microb. Technol 37 : 300 ~ 308
Chambers RW / 1957 / Nucleoside polyphosphates. V.1 Syntheses of guanosine 5’-di- and triphosphates / J. Am. Chem. Soc 79 : 3752 ~ 3755
Chen RR / 2007 / Permeability issues in whole-cell bioprocesses and cellular membrane engineering / Appl. Microbiol. Biotechnol 74 : 730 ~ 738
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