广西植物
廣西植物
엄서식물
GUIHAIA
2013年
1期
118-121
,共4页
烟草%吡哆胺‐丙酮酸转氨酶%纯化%性质分析
煙草%吡哆胺‐丙酮痠轉氨酶%純化%性質分析
연초%필치알‐병동산전안매%순화%성질분석
tobacco%pyridoxamine‐pyruvate aminotransferase%purification%properties
通过冷冻干燥、DEAE‐Sepharose Fast Flow阴离子交换柱层析、SephadexG‐100凝胶过滤柱层析, SP葡聚糖凝胶C‐25阳离子交换柱层析等分离纯化技术, 对烟草吡哆胺‐丙酮酸转氨酶进行分离纯化. 采用苯肼衍生化方法检测活性, 并对其基本酶学性质进行分析. 结果显示:该酶被纯化了92. 34倍;最适温度为70℃, 最适pH为9. 0. 在pH7. 0~9. 0内稳定且热稳定性较好, 80℃保温3h仍有51. 55%的酶活力;在最适反应条件下, 测得反应底物吡哆胺和丙酮酸的Km值分别为6. 337mmol?L‐1和0. 867mmol?L‐1. 该结果为进一步研究烟草体内VB6代谢机制奠定了基础.
通過冷凍榦燥、DEAE‐Sepharose Fast Flow陰離子交換柱層析、SephadexG‐100凝膠過濾柱層析, SP葡聚糖凝膠C‐25暘離子交換柱層析等分離純化技術, 對煙草吡哆胺‐丙酮痠轉氨酶進行分離純化. 採用苯肼衍生化方法檢測活性, 併對其基本酶學性質進行分析. 結果顯示:該酶被純化瞭92. 34倍;最適溫度為70℃, 最適pH為9. 0. 在pH7. 0~9. 0內穩定且熱穩定性較好, 80℃保溫3h仍有51. 55%的酶活力;在最適反應條件下, 測得反應底物吡哆胺和丙酮痠的Km值分彆為6. 337mmol?L‐1和0. 867mmol?L‐1. 該結果為進一步研究煙草體內VB6代謝機製奠定瞭基礎.
통과냉동간조、DEAE‐Sepharose Fast Flow음리자교환주층석、SephadexG‐100응효과려주층석, SP포취당응효C‐25양리자교환주층석등분리순화기술, 대연초필치알‐병동산전안매진행분리순화. 채용분정연생화방법검측활성, 병대기기본매학성질진행분석. 결과현시:해매피순화료92. 34배;최괄온도위70℃, 최괄pH위9. 0. 재pH7. 0~9. 0내은정차열은정성교호, 80℃보온3h잉유51. 55%적매활력;재최괄반응조건하, 측득반응저물필치알화병동산적Km치분별위6. 337mmol?L‐1화0. 867mmol?L‐1. 해결과위진일보연구연초체내VB6대사궤제전정료기출.
@@@@The pyridoxamine‐pyruvate aminotransferase was purified from the tobacco leaf by freeze‐drying ,DEAE Sepharose Fast Flow ion exchange chromatography ,Sephadex G‐100 gel filtration and SP Sephadex C‐25 ion ex‐change chromatography .The enzymatic activity and properties were investigated with phenyl hydrazine method .The results showed that 92 .34‐fold purification was obtained ;the enzyme had an optimum temperature at 70 ℃ and pH at 9.0 ,and it was stable at pH7 .0 -9 .0 ;the enzyme had good thermal stability which remained about 51 .55% of its activity after being treated at 80 ℃ for 3 h ;under optimal conditions ,the Km values for pyridoxamine and pyruvate were 6 .337 mmol ? L‐1 and 0 .867 mmol ? L‐1 .The results provided basis for the metabolic mechanism of VB 6 in tobacco plants .
