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논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
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- 한국미생물생명공학회 Journal of Microbiology and Biotechnology Journal of Microbiology and Biotechnology 제14권 제2호
- 발행연도
- 2004.1
- 수록면
- 342 - 349 (8page)
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초록· 키워드
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A gene encoding inulin-degrading enzyme ofBacillus sp. snu-7 with ORF of 1536 nucleotides was cloned.And it was overexpressed as His-tagged protein in E. coliBL21(DE3) pLysS using pRSET B vector containing matureenzyme sequence. Maximum enzyme production was achievedby IPTG (0.1 mM) induction at OD600 1.2 and 30o C, followedby 6 h incubation. The expressed protein purified throughimmobilized metal affinity chromatography showed molecularmass of 60 kDa on SDS-PAGE. Results of thin-layerchromatography using inulin as a substrate showed the enzymeto be an exotype inulinase capable of producing only monomericfructose as a product. Km and kcat for the hydrolyses of inulinand sucrose were 2.28±0.08 mM and 358.05± 20.38 min- 1,and 22.02±0.41 mM and 4619.11±215.12 min-1, respectively.Optimal activity of the exoinulinase occurred at pH 7.0 and50o C.
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참고문헌 (9)
참고문헌 신청
Arand, M / 2002 / Purification,characterization, gene cloning and preliminary X-ray data ofthe exo-inulinase from Aspergillus awamori 362 : 131 ~ 135
Bradford / 1976 / A rapid and sensitive method for thequantitation of microgram of protein utilizing the principleof protein dye-binding : 248 ~ 254
Cohen / 1972 / Nonchromosomal antibiotic resistance in bacteria Genetictransformation of Escherichia coli by R-factor DNA : 2110 ~ 69 2114
Copeland / 2000 / Enzymes: A Practical Introduction toStructure, Mechanism, and Data Analysis. 2nd Ed
Fuchs / 1993 / Production and utilization of inulin Utilization of inulin Science and Technology of Fructans : 325 ~ 346
Bradford / 1976 / A rapid and sensitive method for thequantitation of microgram of protein utilizing the principleof protein dye-binding : 248 ~ 254
Cohen / 1972 / Nonchromosomal antibiotic resistance in bacteria Genetictransformation of Escherichia coli by R-factor DNA : 2110 ~ 69 2114
Copeland / 2000 / Enzymes: A Practical Introduction toStructure, Mechanism, and Data Analysis. 2nd Ed
Fuchs / 1993 / Production and utilization of inulin Utilization of inulin Science and Technology of Fructans : 325 ~ 346
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