Measuring Lactase Enzymatic Activity in the Teaching Lab.
Measuring Lactase Enzymatic Activity in the Teaching Lab.
复制标题
在教学实验室中测量乳糖酶活性。
DOI：
10.3791/54377
复制
发表时间：
2018-08-06
期刊：
影响因子：
--
通讯作者：
Lewis PA
中科院分区：
文献类型：
--
作者：
Leksmono CS;Manzoni C;Tomkins JE;Lucchesi W;Cottrell G;Lewis PA
了解酶是如何工作的，并将其与真实的生命实例联系起来，对于生物和生物医学科学的广泛本科学位至关重要。这个易于遵循的协议是为一年级的本科药学专业的学生，并提供了一个入门级的介绍酶反应和酶分析的分析程序。选择的酶是乳糖酶，因为这代表了与人类疾病/药学实践相关的市售酶的实例。从膳食补充剂片剂中提取乳糖酶，并使用基于乳糖酶人工底物（邻硝基苯酚-β-D-吡喃半乳糖苷，ONPG）水解的比色测定法进行评估。通过420 nm处吸光度的变化测量乳糖酶水解ONPG后邻硝基苯酚的释放，并通过在冰上、室温和37 ℃下进行反应来评估温度对酶促反应的影响。通过在不同条件下和使用不同试剂评估酶活性，可以使用该方案实施更高级的分析。
Understanding how enzymes work, and relating this to real life examples, is critical to a wide range of undergraduate degrees in the biological and biomedical sciences. This easy to follow protocol was developed for first year undergraduate pharmacy students and provides an entry-level introduction to enzyme reactions and analytical procedures for enzyme analysis. The enzyme of choice is lactase, as this represents an example of a commercially available enzyme relevant to human disease/pharmaceutical practice. Lactase is extracted from dietary supplement tablets, and assessed using a colorimetric assay based upon hydrolysis of an artificial substrate for lactase (ortho-nitrophenol-beta-D-galactopyranoside, ONPG). Release of ortho-nitrophenol following the hydrolytic cleavage of ONPG by lactase is measured by a change in absorbance at 420 nm, and the effect of the temperature on the enzymatic reaction is evaluated by carrying out the reaction on ice, at room temperature and at 37 °C. More advanced analysis can be implemented using this protocol by assessing the enzyme activity under different conditions and using different reagents.
登录
查看更多内容
影响因子：
9.8
作者：
Enattah, Nabil Sabri;Jensen, Tine G. K.;Peltonen, Leena
通讯作者：
Peltonen, Leena
影响因子：
2.9
作者：
Juers, DH;Heightman, TD;Matthews, BW
通讯作者：
Matthews, BW
影响因子：
1.4
作者：
Jittam, Piyachat;Boonsiri, Patcharee;Panijpan, Bhinyo
通讯作者：
Panijpan, Bhinyo
影响因子：
3
作者：
RUSSO, SF;MOOTHART, L
通讯作者：
MOOTHART, L
影响因子：
4.1
作者：
Krebs, HA;Salvin, E;Johnson, WA
通讯作者：
Johnson, WA