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학술연구/단체지원/교육 등 연구자 활동을 지속하도록 DBpia가 지원하고 있어요.
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논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 저널정보
- 한국미생물생명공학회 Journal of Microbiology and Biotechnology Journal of Microbiology and Biotechnology 제22권 제3호
- 발행연도
- 2012.1
- 수록면
- 292 - 300 (9page)
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We report the expression, purification, and characterization of L-asparaginase (AnsA) from Rhizobium etli. The enzyme was purified to homogeneity in a single-step procedure involving affinity chromatography, and the kinetic parameters Km, Vmax, and kcat for L-asparagine were determined. The enzymatic activity in the presence of a number of substrates and metal ions was investigated. The molecular mass of the enzyme was 47 kDa by SDS-PAGE. The enzyme showed a maximal activity at 50oC, but the optimal temperature of activity was 37oC. It also showed maximal and optimal activities at pH 9.0. The values of Km, Vmax,kcat, and kcat/Km were 8.9 ± 0.967 × 10-3 M, 128 ± 2.8 U/mg protein, 106 ± 2 s-1, and 1.2 ± 0.105 × 104M-1s-1, respectively.
The L-asparaginase activity was reduced in the presence of Mn2+, Zn2+, Ca2+, and Mg2+ metal ions for about 52% to 31%. In addition, we found that NH4+, L-Asp, D-Asn, and β-aspartyl-hydroxamate in the reaction buffer reduced the activity of the enzyme, whereas L-Gln did not modify its enzymatic activity. This is the first report on the expression and characterization of the L-asparaginase (AnsA) from R. etli. Phylogenetic analysis of asparaginases reveals an increasing group of known sequences of the Rhizobialtype asparaginase II family.
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참고문헌 (23)
참고문헌 신청
[학술지(정기간행물
Abdel-Fattah, Y. R. / 2002 / L-Asparaginase production by Pseudomonas aeruginosa in solid-state culture: Evaluation and optimization of culture conditions using factorial des
Ashworth, L. A. / 1974 / Comparison of the L-asparaginases from Escherichia coli and Erwinia carotovora as immunosuppressants / Cancer Res 34 : 1353 ~ 1359
Balcao, V. M. / 2001 / Structural and functional stabilization of L-asparaginase via multisubunit immobilization onto highly activated supports / Biotechnol. Prog 17 : 537 ~
Bao, K. / 2001 / Terminal proteins essential for the replication of linear plasmids and chromosomes in Streptomyces / Genes Dev 15 : 1518 ~ 1527
Abdel-Fattah, Y. R. / 2002 / L-Asparaginase production by Pseudomonas aeruginosa in solid-state culture: Evaluation and optimization of culture conditions using factorial des
Ashworth, L. A. / 1974 / Comparison of the L-asparaginases from Escherichia coli and Erwinia carotovora as immunosuppressants / Cancer Res 34 : 1353 ~ 1359
Balcao, V. M. / 2001 / Structural and functional stabilization of L-asparaginase via multisubunit immobilization onto highly activated supports / Biotechnol. Prog 17 : 537 ~
Bao, K. / 2001 / Terminal proteins essential for the replication of linear plasmids and chromosomes in Streptomyces / Genes Dev 15 : 1518 ~ 1527
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