广西科学
廣西科學
엄서과학
GUANGXI SCIENCES
2014年
2期
115-118
,共4页
滕昆%张吉福%牛福星%杜丽琴%韦宇拓
滕昆%張吉福%牛福星%杜麗琴%韋宇拓
등곤%장길복%우복성%두려금%위우탁
脂肪酶%酶活测定%终止反应%抑制效果
脂肪酶%酶活測定%終止反應%抑製效果
지방매%매활측정%종지반응%억제효과
lipase%activity assay%termination re-action%inhibiting effect
【目的】寻找一种准确有效的脂肪酶活力测定中的终止反应方法，以进行脂肪酶酶学性质研究。【方法】比较分析前人研究脂肪酶常用的终止方法（无水乙醇法、EDTA 法、三氯乙酸法、碳酸钠法、沸水浴法）对酶活力的抑制效果和吸光值的稳定性。【结果】无水乙醇法对酶活的抑制率为97．56％，终止反应后5min 内净吸光值减少5％；EDTA 法对酶活抑制率为32．94％，终止反应后10min 内净吸光值增加5％；三氯乙酸法对酶活的抑制率为99．46％，终止反应后10min 内净吸光值增加了0．47％；碳酸钠法对酶活抑制率为76．44％，终止反应后10min 内净吸光值增加了0．53％；沸水浴法对酶活抑制率为99．51％，终止反应后10min 内净吸光值减少0·77％，但沸水浴会导致剩余底物的大量分解。【结论】三氯乙酸法是一种准确有效的终止脂肪酶活力测定反应的方法，可用于脂肪酶酶学性质的研究。
【目的】尋找一種準確有效的脂肪酶活力測定中的終止反應方法，以進行脂肪酶酶學性質研究。【方法】比較分析前人研究脂肪酶常用的終止方法（無水乙醇法、EDTA 法、三氯乙痠法、碳痠鈉法、沸水浴法）對酶活力的抑製效果和吸光值的穩定性。【結果】無水乙醇法對酶活的抑製率為97．56％，終止反應後5min 內淨吸光值減少5％；EDTA 法對酶活抑製率為32．94％，終止反應後10min 內淨吸光值增加5％；三氯乙痠法對酶活的抑製率為99．46％，終止反應後10min 內淨吸光值增加瞭0．47％；碳痠鈉法對酶活抑製率為76．44％，終止反應後10min 內淨吸光值增加瞭0．53％；沸水浴法對酶活抑製率為99．51％，終止反應後10min 內淨吸光值減少0·77％，但沸水浴會導緻剩餘底物的大量分解。【結論】三氯乙痠法是一種準確有效的終止脂肪酶活力測定反應的方法，可用于脂肪酶酶學性質的研究。
【목적】심조일충준학유효적지방매활력측정중적종지반응방법，이진행지방매매학성질연구。【방법】비교분석전인연구지방매상용적종지방법（무수을순법、EDTA 법、삼록을산법、탄산납법、비수욕법）대매활력적억제효과화흡광치적은정성。【결과】무수을순법대매활적억제솔위97．56％，종지반응후5min 내정흡광치감소5％；EDTA 법대매활억제솔위32．94％，종지반응후10min 내정흡광치증가5％；삼록을산법대매활적억제솔위99．46％，종지반응후10min 내정흡광치증가료0．47％；탄산납법대매활억제솔위76．44％，종지반응후10min 내정흡광치증가료0．53％；비수욕법대매활억제솔위99．51％，종지반응후10min 내정흡광치감소0·77％，단비수욕회도치잉여저물적대량분해。【결론】삼록을산법시일충준학유효적종지지방매활력측정반응적방법，가용우지방매매학성질적연구。
[Objective]Searching an accurate and effective termination reaction method in lipase activity assay for studying lipase enzymatic characters.[Methods]Comparative analyses were conducted in inhibiting effect and result stability of termination methods(absolute ethanol,ED-TA,trichloroacetic acid,sodium carbonate and boiling water bath),which predecessors used generally in lipase activity assay.[Results]Inhibition ratio of termination by absolute ethanol was 97.56%,and net absorbance reduced by 5% within 5min after termination;Inhibition ratio of terminaion by EDTA was 32.94%,and net absorbance increased by 5% within 10min after termination;Inhibition ratio of termination by trichloroacetic acid was 99.46%,and coloration net absorbance increased by 0.47% within 10min after termination;Inhibition ratio of termina-tion by sodium carbonate was 76.44%,and net absorbance increased by 0.53% within 10min after termination;Inhibition ratio of termination by boiling water was 99.51%,and net absor-bance reduced by 0.77% within 10min after termination, but it would result in the decomposition of most remaining substrates.[Conclusion]The termination method of trichlo-roacetic acid is an accurate and effective one in li-pase activity assay,and can be adopt in analyzing lipase enzymatic characters.
