Name: hot biological catalysis: isothermal titration calorimetry to characterize enzymatic reactions
Uploaded: 2014-04-04T13:45:00
Description: Watch this Scientific Journal Video about Hot Biological Catalysis: Isothermal Titration Calorimetry to Characterize Enzymatic Reactions at JoVE.com

该专用肥产品公司（SFP）是公认的提供必要的资金用于这项研究。

1。样品的准备<ol><li>制备2毫升酶溶液和0.5ml的每个实验运行的底物溶液。在具有相同的组合物到基片加入过程中尽量减少稀释和混合的热缓冲液稀释浓缩的储备溶液酶和底物。 <ol><li>选择的缓冲液条件是足够的，以防止试验过程中pH值的变化。例如，20mM的HEPES pH值为7是足够的测量值在中性pH值。 注意：如果涉及质子交换，所使用的缓冲液的质子化焓必须考虑的，因为它们会影响该反?...

<ol>
	<li>Leavitt, S., Freire, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Direct+measurement+of+protein+binding+energetics+by+isothermal+titration+calorimetry.">Direct measurement of protein binding energetics by isothermal titration calorimetry.</a> Curr. Opin. Struct. Biol. 11, 560-566 (2001).</li><li>Ladbury, J. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Application+of+isothermal+titration+calorimetry+in+the+biological+sciences:+things+are+heating+up!+BioTechniques.">Application of isothermal titration calorimetry in the biological sciences: things are heating up! BioTechniques.</a> 37, 885-887 (2004).</li><li>Zambelli, B., Bellucci, M., Danielli, A., Scarlato, V., Ciurli, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Ni2++binding+properties+of+Helicobacter+pylori+NikR.">The Ni2+ binding properties of Helicobacter pylori NikR.</a> Chem. Commun. , 3649-3651 (2007).</li><li>Zambelli, B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High-affinity+Ni2++binding+selectively+promotes+binding+of+Helicobacter+pylori+NikR+to+its+target+urease+promoter.">High-affinity Ni2+ binding selectively promotes binding of Helicobacter pylori NikR to its target urease promoter.</a> J. Mol. Biol. 383, 1129-1143 (2008).</li><li>Duff, M. R., Grubbs, J., Howell, E. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isothermal+titration+calorimetry+for+measuring+macromolecule-ligand+affinity.">Isothermal titration calorimetry for measuring macromolecule-ligand affinity.</a> J. Vis. Exp. , (2011).</li><li>Ghai, R., Falconer, R. J., Collins, B. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Applications+of+isothermal+titration+calorimetry+in+pure+and+applied+research--survey+of+the+literature+from+2010.">Applications of isothermal titration calorimetry in pure and applied research--survey of the literature from 2010.</a> J. Mol. Recognit. 25, 32-52 (2012).</li><li>Todd, M. J., Gomez, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Enzyme+kinetics+determined+using+calorimetry:+a+general+assay+for+enzyme+activity.">Enzyme kinetics determined using calorimetry: a general assay for enzyme activity.</a> Anal. Biochem. 296, 179-187 (2001).</li><li>Bianconi, M. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Calorimetry+of+enzyme-catalyzed+reactions.">Calorimetry of enzyme-catalyzed reactions.</a> Biophys. Chem. 126, 59-64 (2007).</li><li>Demarse, N. A., Killian, M. C., Hansen, L. D., Quinn, C. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Determining+enzyme+kinetics+via+isothermal+titration+calorimetry.">Determining enzyme kinetics via isothermal titration calorimetry.</a> Methods Mol. Biol. 978, 21-30 (2013).</li><li>Michaelis, L., Menten, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Die+kinetik+der+invertinwirkung.">Die kinetik der invertinwirkung.</a> Biochem. Z. 49, 333-369 (1913).</li><li>Walker, J., Wilson, K., Walker, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Principle and techniques of practical biochemistry. , 312-356 (2000).</li><li>Ciurli, S., Sigel, A., Sigel, H., Sigel, R. K. O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Nickel and its surprising impact in nature. 2, 241-278 (2007).</li><li>Zambelli, B., Musiani, F., Benini, S., Ciurli, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chemistry+of+Ni2++in+urease:+sensing,+trafficking,+and+catalysis.">Chemistry of Ni2+ in urease: sensing, trafficking, and catalysis.</a> Acc. Chem. Res. 44, 520-530 (2011).</li><li>Zonia, L. E., Stebbins, N. E., Polacco, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Essential+role+of+urease+in+germination+of+nitrogen-limited+Arabidopsis+thaliana+seeds.">Essential role of urease in germination of nitrogen-limited Arabidopsis thaliana seeds.</a> Plant Physiol. 107, 1097-1103 (1995).</li><li>Follmer, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Insights+into+the+role+and+structure+of+plant+ureases.">Insights into the role and structure of plant ureases.</a> Phytochemistry. 69, 18-28 (2008).</li><li>Sumner, J. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+isolation+and+crystallization+of+the+enzyme+urease.">The isolation and crystallization of the enzyme urease.</a> J. Biol. Chem. 69, 435-441 (1926).</li><li>Krajewska, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ureases+I.+Functional,+catalytic+and+kinetic+properties:+A+review.">Ureases I. Functional, catalytic and kinetic properties: A review.</a> J. Mol. Cat. B. 59, 9-21 (2009).</li><li>Callahan, B. P., Yuan, Y., Wolfenden, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+burden+borne+by+urease.">The burden borne by urease.</a> J. Am. Chem. Soc. 127, 10828-10829 (2005).</li><li>Krajewska, B., van Eldik, R., Brindell, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Temperature-+and+pressure-dependent+stopped-flow+kinetic+studies+of+jack+bean+urease.+Implications+for+the+catalytic+mechanism.">Temperature- and pressure-dependent stopped-flow kinetic studies of jack bean urease. Implications for the catalytic mechanism.</a> J. Biol. Inorg. Chem. 17, 1123-1134 (2012).</li><li>Hausinger, R. P., Karplus, P. A., Huber, R., Poulos, T., Wieghardt, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Handbook of Metalloproteins. , 867-879 (2001).</li><li>Goldberg, R., Kishore, N., Lennen, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Thermodynamic+quantities+for+the+ionization+reactions+of+buffers.">Thermodynamic quantities for the ionization reactions of buffers.</a> J. Phys. Chem. Ref. Data. 31, 231-370 (2002).</li><li>Baumann, M. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Advantages+of+isothermal+titration+calorimetry+for+xylanase+kinetics+in+comparison+to+chemical-reducing-end+assays.">Advantages of isothermal titration calorimetry for xylanase kinetics in comparison to chemical-reducing-end assays.</a> Anal. Biochem. 410, 19-26 (2011).</li><li>Noske, R., Cornelius, F., Clarke, R. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Investigation+of+the+enzymatic+activity+of+the+Na+,K+-ATPase+via+isothermal+titration+microcalorimetry.">Investigation of the enzymatic activity of the Na+,K+-ATPase via isothermal titration microcalorimetry.</a> Biochim. Biophys. Acta. 1797, 1540-1545 (2010).</li><li>Harmon, K. M., Niemann, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+competitive+inhibition+of+the+urease-catalyzed+hydrolysis+of+urea+by+phosphate.">The competitive inhibition of the urease-catalyzed hydrolysis of urea by phosphate.</a> J. Biol. Chem. 177, 601-605 (1949).</li><li>Benini, S., Rypniewski, W. R., Wilson, K. S., Ciurli, S., Mangani, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Structure-based+rationalization+of+urease+inhibition+by+phosphate:+novel+insights+into+the+enzyme+mechanism.">Structure-based rationalization of urease inhibition by phosphate: novel insights into the enzyme mechanism.</a> J. Biol. Inorg. Chem. 6, 778-790 (2001).</li><li>Segel, I. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Enzyme kinetics: behavior and analysis of rapid equilibrium and steady-state enzyme systems. , (1993).</li></ol>

ITC的意义，研究酶的活性就现有的方法 除了其经典应用研究的结合平衡，等温滴定量热法提供了可靠的和快速的方法，使用的反应热作为探针来表征溶液中的​​酶反应，而不需要对系统的修改或标记。动力学参数K 猫和K M分别通过一组时间过程实验，其中的产物的形成（或底物消耗）在连续或不连续测定法进行监测，通常获得的?...

定量测定生化反应提供了深入了解生物过程在生命的基础。量热法提供了一种无标记的方法来定量地表征溶液中的​​几乎所有的化学反应。这种技术测量热释放或吸收随着时间的推移，因此是一种普遍的检测系统和一个非常方便的方法来量化反应的分子（ 即结合热力学），以及测量的反应速率（ 即动力学）的量。特别是，等温滴定量热法（ITC）已被采纳为首选方法来表征生物分子平衡的热力学，涉及蛋白质-配体，蛋白质-蛋白质，蛋白质-金属离子与蛋白-DNA相互作用1-6。此外，国际贸易中心提供动力学信息的能力，使其成为一个非常强大的系统来衡量酶的催化作用，虽然这个应用程序的潜力仍然低估了7-9。 米氏方程10是酶反应的定量描述，因为它提供了反应速率和底物浓度之间的关系，这取决于2动力学参数：米氏常数（K M）和催化速率常数（k 猫 ） 。 第 k 猫 / K M比值被称为一种酶的催化效率。在实践中，对于特定的反应测定K M和 k 猫提供催化的完整描述。 在一个典型的酶反应（ 图1），在衬底（S）相互作用的酶（E）形成的酶-底物（ES）的配合物，其随后被激活进入过渡状态（ES *）。后者被转化成酶产品（EP）的复杂的最终解离。这些步S是由以下的反应说明。 <img fo:content-width="2in" src="/files/ftp_upload/51487/51487eq1.jpg" width="197" /> （1） 其中，k 1是速率常数为ES复合物中，k -1的形成是速率常数为ES复合物的解离，而K是猫的催化速率常数或周转数。 根据米氏方程10中 ，该反应的速率可以计算为： <img fo:content-width="1.5in" src="/files/ftp_upload/51487/51487eq2.jpg" width="144" /> （2） 其中K M =（K-1 + K 猫 ）/ k1 和 k2 的猫 = V 最大 / [E]， 用 v max均当所有的酶结合到基体达到的最大速度。 等温滴定热量计是在本研究中用于表征尿素的酶水解仪器。该仪器由含有2杜撰形细胞（ 图1）的绝热屏蔽的。这些被连接到外部以窄接入管中。样品池（ 约 1.4ml）中装入与酶溶液，同时基准单元通常被充满水或与用于分析的溶剂。一种旋转式注射器用长针和相连的搅拌桨，通常含有约 0.3ml的底物溶液，装在样品池。一种热电装置测量温度的样品和参考单元之间的差异，采用了“小区反馈网络”，它通过增加或减少热保持这种差异为零。在实验过程中，衬底被注入到在一个恒定的温度选择的酶溶液。当ENzymatic发生反应，热释放或吸收的量是成比例的转换成产物的分子底物分子的数量。另外，热气流的速度直接相关，该反应的速率。测得的数据，表现为发热微量的（ 图1）的偏差从初始基线，表示热功率（μcal/秒）供给由仪器的样品池，其正比于所述试样盒中发生的热流量随着时间的推移。 <img alt="图1" fo:content-width="5in" src="/files/ftp_upload/51487/51487fig1highres.jpg" width="500" /> 图1。的等温滴定量热仪来研究酶反应的示意图。发生后的衬底（在注射器）的滴定到酶溶液（在样品池）的结果在该疗法的一个变化的酶促反应由量热仪，需要保持样品池和参比电极常数之间的温度差正常释放动力。 <a href="https://www.jove.com/files/ftp_upload/51487/51487fig1highres.jpg" target="_blank">点击这里查看大图。</a> 总体上，热变化（Q）是正比于反应（ΔH）和生成（n）的，而这又是通过总体积时代赋予的浓度产物的摩尔数的摩尔焓： <img fo:content-width="2in" src="/files/ftp_upload/51487/51487eq3.jpg" width="200" /> （3） 该产物的形成随时间（DP / dt）的，对应于反应速率，因此，可以与以上通过关系在同一时间（的dQ / dt）的产生的热量： <img fo:content-width="2in" src="/files/ftp_upload/51487/51487eq4.jpg" width="150" /> （4） 根据这个公式，以便获得一个米氏绘制它需要测量ⅰ）的总摩尔焓ΔH，以及ii）的热流动的dQ / dt的在不同的底物浓度。通常，这是在两个不同的实验进行：在第一个实验中（ 方法1，M1），其中所述衬底中注入酶溶液，并加热为完整的底物转化的测量;在第二个实验（ 方法2，M2），底物的多次注射执行和热产生率的测量是在不同的底物浓度。这两组数据的数据足以计算得出的动力学参数K M和 k 的猫 。 在本文章中，一般的协议来确定动力学参数，使用国际贸易中心进行酶促反应描述。我们通过刀豆尿素施用的方法，以尿素水解本身作为参照系。使用这种方法得到的结果和文献报道的数据之间的吻合证明了这种方法的可靠性。

<table border="0" cellpadding="0" cellspacing="0" width="796">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:245px;">HEPES</td>
			<td style="width:111px;">Sigma</td>
			<td style="width:119px;">H3375</td>
			<td style="width:323px;">dissolving in water and adjusting pH with NaOH</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:245px;">TRIZMA-Base</td>
			<td style="width:111px;">Sigma</td>
			<td style="width:119px;">T1503</td>
			<td>dissolving in water and adjusting pH with HCl</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:245px;">Sodium dihydrogen phosphate</td>
			<td style="width:111px;">Riedel-de-Haen</td>
			<td style="width:119px;">4270</td>
			<td>dissolving in water</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:245px;">Sodium phosphate dibasic</td>
			<td style="width:111px;">Riedel-de-Haen</td>
			<td style="width:119px;">30427</td>
			<td>dissolving in water</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:245px;">Urea</td>
			<td style="width:111px;">Sigma</td>
			<td style="width:119px;">U4128</td>
			<td>dissolving in water at 40 &#176;C</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:245px;">Canavalia ensiformis urease (type C-3)</td>
			<td style="width:111px;">Sigma</td>
			<td style="width:119px;">U0251</td>
			<td>dissolving in 20 mM HEPES pH 7 and stored at -80 &#176;C</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:245px;"></td>
			<td style="width:111px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:245px;">VP-ITC on Origin 7.0</td>
			<td style="width:111px;">MicroCal (GE Healthcare)</td>
			<td style="width:119px;">SYS13901</td>
			<td>instrument&#160;</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:245px;">VPViewer2000 1.30.00 on Origin 7.0</td>
			<td style="width:111px;">MicroCal (GE Healthcare)</td>
			<td style="width:119px;"></td>
			<td>data acquisition software supplied with the instrument</td>
		</tr>
	</tbody>
</table>

hot biological catalysis: isothermal titration calorimetry to characterize enzymatic reactions

等温滴定量热法（ITC）是一个良好描述的技术，其测量过程中的化学反应的热释放或吸收，用它作为一个内在探头几乎以表征每一个化学过程。如今，这种技术被广泛地应用，以确定生物分子的结合平衡的热力学参数。此外，ITC已经被证明是能够直接测量酶促反应的动力学和热力学参数（K 猫 ，K M，△H），即使该应用程序仍是开发不足的。作为热变化过程中酶催化自发地发生，ITC，不需要该系统的下分析的任何修改或标签，并且可以在溶液中进行。此外，该方法需要的材料少量。这些特性使得德祥一个非常宝贵的，强大的和独特的工具来研究酶动力学的几个应用，如，例如，药物发现。 K值猫和尿素由刀豆 （刀豆）脲酶酶水解的K M。进行反应的内在摩尔焓（ΔHint）的计算。由此得到的值与文献中报道的先前的数据相一致，这表明该方法的可靠性。

脲酶（EC 3.5.1.5;尿素水解酶）是古细菌，细菌，单细胞真核生物和植物中发现一个多亚基含镍酶。这种蛋白作用于有机氮矿化的最后步骤，催化尿素水解为氨和氨基甲酸盐，其自发地分解，得到氨和碳酸氢钠（ 公式6）12的第二分子。 <img fo:content-width="4in" src="/files/ftp_upload/51487/51487eq6.jpg" width="450" /> （6） 该反应导致的环境中，这是负?...

Watch this Scientific Journal Video about Hot Biological Catalysis: Isothermal Titration Calorimetry to Characterize Enzymatic Reactions at JoVE.com

Hot Biological Catalysis: Isothermal Titration Calorimetry to Characterize Enzymatic Reactions

等温滴定量热法测量热流释放或吸收的化学反应。该方法可用于定量酶催化。在本文中，该协议有助于建立，运行试验和数据分析通常被描述，并应用于酶水解尿素由刀豆脲酶的特性。

热生物催化：等温滴定量热法表征酶促反应

Isothermal titration calorimetry (ITC) is a well-described technique that measures the heat released or absorbed during a chemical reaction, using it as ...

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Retropinacol/Cross-pinacol Coupling Reactions - A Catalytic Access to 1,2-Unsymmetrical Diols

Retropinacol /跨频哪醇偶联反应 - 催化访问1,2  - 非对称二醇

Unsymmetrical 1,2-diols are hardly accessible by reductive pinacol coupling processes. A successful execution of such a transformation is bound to a clear ...

Mizoroki-Heck Cross-coupling Reactions Catalyzed by Dichloro{bis[1,1',1''-(phosphinetriyl)tripiperidine]}palladium Under Mild Reaction Conditions

Mizoroki-Heck Cross-coupling Reactions Catalyzed by Dichloro{bis[1,1',1''-phosphinetriyltripiperidine]}palladium Under Mild Reaction Conditions

Mizoroki-哎呀交叉偶联反应由二氯{双[1,1&#39;，1&#39;&#39;  - （phosphinetriyl）tripiperidine]}钯催化在温和的反应条件

Dichloro-bis(aminophosphine) complexes of palladium with the general formula of [(P{(NC5H10)3-n(C6H11)n})2Pd(Cl)2] (where n = 0-2), belong to a new family ...

Light-driven Enzymatic Decarboxylation

光驱动脱羧酶

Oxidoreductases belong to the most-applied industrial enzymes. Nevertheless, they need external electrons whose supply is often costly and challenging. ...

Dissolution Dynamic Nuclear Polarization Instrumentation for Real-time Enzymatic Reaction Rate Measurements by NMR

溶出动态核极化仪器用NMR实时酶反应速率测量

The main limitation of NMR-based investigations is low sensitivity. This prompts for long acquisition times, thus preventing real-time NMR measurements of ...

A Protocol for Safe Lithiation Reactions Using Organolithium Reagents

使用有机锂试剂的一个协议安全的锂化反应

Organolithium reagents are powerful tools in the synthetic chemist&#39;s toolbox. However, the extreme pyrophoric nature of the most reactive reagents ...

Enzymatic Cascade Reactions for the Synthesis of Chiral Amino Alcohols from L-lysine

酶联反应合成 l-赖氨酸手性氨基醇的研究

Amino alcohols are versatile compounds with a wide range of applications. For instance, they have been used as chiral scaffolds in organic synthesis. ...

Employing Pressurized Hot Water Extraction (PHWE) to Explore Natural Products Chemistry in the Undergraduate Laboratory

Employing Pressurized Hot Water Extraction PHWE to Explore Natural Products Chemistry in the Undergraduate Laboratory

利用加压热水提取 (phwe) 在本科实验室中探索天然产物化学

A recently developed pressurized hot water extraction (PHWE) method which utilizes an unmodified household espresso machine to facilitate natural products ...

Imine Metathesis by Silica-Supported Catalysts Using the Methodology of Surface Organometallic Chemistry

利用表面有机金属化学方法，利用硅支持的催化剂进行Imine金属分析

With this protocol, a well-defined singlesite silica-supported heterogeneous catalyst [(&#8801;Si-O-)Hf(=NMe)(&#951;1-NMe2)] is designed and prepared ...

Enzymatic Synthesis of Epoxidized Metabolites of Docosahexaenoic, Eicosapentaenoic, and Arachidonic Acids

多克沙六烯、环氧体和阿拉奇酮酸的环氧代谢物酶合成

The epoxidized metabolites of various polyunsaturated fatty acids (PUFAs), termed epoxy fatty acids, have a wide range of roles in human physiology. These ...