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논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 저널정보
- 한국응용생명화학회 Journal of applied biological chemistry Journal of applied biological chemistry 제54권 제4호
- 발행연도
- 2011.1
- 수록면
- 231 - 237 (7page)
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A gene encoding a putative alanine racemase in Xanthomonas. oryzae pv. oryzae was cloned, expressed and characterized. Expression of the cloned gene was performed in Escherichia coli BL21(DE3)pLys using a pET-21(a) vector harbouring $6{\times}histidine$ tag. Purification of the recombinant alanine racemase by affinity chromatography resulted in major one band by sodium dodecyl sulfate polyacryl amide gel electrophoresis analysis, showing about 45 kDa of molecular weight. The alanine racemase gene, cloned in this experiment, appears to be constitutively expressed in X. oryzae, as analyzed by reverse transcriptase polymerase chain reaction. The enzyme was the most active toward L-alanine and secondly D-alanine, showing a racemic reaction, thus the enzyme is considered as an alanine racemase. The enzyme was considerably activated by addition of pyridoxal-5-phosphate (PLP), showing that 75% increase in activity was observed at 0.3 mM, compared with control. D-Cysteine as well as L-cysteine significantly inhibited the enzyme activity. The inhibitions by cysteines were more prominent in the absence of PLP, showing 9 and 5% of control activity at 2 mM of addition, respectively. The enzyme was the most active at pH 8.0 and more stable at alkaline pHs than acidic pH condition.
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참고문헌 (33)
참고문헌 신청
Badet B / 1985 / Purification of an alanine racemase from Streptococcus faecalis and analysis of its inactivation by (1-aminoethyl) phosphonic acid enantiomers / Biochemistry 24:1333~1341
Bradford MM / 1976 / A rapid sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein dye binding / Anal Biochem 72:248~254
Buell MV / 1960 / Reaction of pyridoxal 5 phosphate with aminothiols / J Am Chem Soc 82:6042~6049
Copie V / 1988 / Inhibition of alanine racemase by alanine phosphonate: detection of an imine linkage to pyridoxal 5’phosphate in the enzyme-inhibitor complex by solidstate N15-nuclear magnetic resonance / Biochemistry 27:4966~4970
Erion MD / 1987 / 1-Aminocyclopropane-phosphonate: Time-dependen inactivation of 1-aminocyclopropanecarboxylate deaminase and Bacillus stearothermophilus alanine racemase by slow dissociation behavior / Biochemistry 26:3417~3425
Bradford MM / 1976 / A rapid sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein dye binding / Anal Biochem 72:248~254
Buell MV / 1960 / Reaction of pyridoxal 5 phosphate with aminothiols / J Am Chem Soc 82:6042~6049
Copie V / 1988 / Inhibition of alanine racemase by alanine phosphonate: detection of an imine linkage to pyridoxal 5’phosphate in the enzyme-inhibitor complex by solidstate N15-nuclear magnetic resonance / Biochemistry 27:4966~4970
Erion MD / 1987 / 1-Aminocyclopropane-phosphonate: Time-dependen inactivation of 1-aminocyclopropanecarboxylate deaminase and Bacillus stearothermophilus alanine racemase by slow dissociation behavior / Biochemistry 26:3417~3425
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