Name: looking outwards: isolation of cyanobacterial released carbohydrate polymers and proteins
Uploaded: 2019-05-27T13:00:00
Description: Watch this Scientific Journal Video about Looking Outwards: Isolation of Cyanobacterial Released Carbohydrate Polymers and Proteins at JoVE.com

这项工作由欧洲发展基金会(FEDER)通过竞争-竞争力和国际化歌剧方案(POCI),葡萄牙2020年,葡萄牙基金通过FCT- 基金会资助a 在POCI-01-01-FEDER-028779项目框架内,在项目POCI-01-0145-FEDER-028779和赠款SFRH/BD/99715/2014(CF)中,技术技术/Ministério da Cióncia,Tecnologia e Ensino 高级版。

1. 氰化物释放碳水化合物聚合物分离
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聚合物分离和去除污染物
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<li>在标准条件下培养蓝藻菌株[例如,在 12 小时光 (50 μE m+ 2s+1) /12 h 暗方案下 30°C,在 150 rpm 下的轨道抖动]。使用标准方案测量生长(例如,730 nm (OD730nm)、叶绿素 a、干重等光学密度,然后根据苯酚-硫酸方法16测量释放多糖的产量。</li>
		<li>将培养层转移到透析膜(分子量截止?...

<ol>
	<li>Pereira, S., 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=Complexity+of+cyanobacterial+exopolysaccharides:+composition,+structures,+inducing+factors+and+putative+genes+involved+in+their+biosynthesis+and+assembly.">Complexity of cyanobacterial exopolysaccharides: composition, structures, inducing factors and putative genes involved in their biosynthesis and assembly.</a> FEMS Microbiology Reviews. 33 (5), 917-941 (2009).</li><li>Kanekiyo, K., 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=Isolation+of+an+Antiviral+Polysaccharide,+Nostoflan,+from+a+Terrestrial+Cyanobacterium,+Nostoc+flagelliforme.">Isolation of an Antiviral Polysaccharide, Nostoflan, from a Terrestrial Cyanobacterium, Nostoc flagelliforme.</a> Journal of Natural Products. 68 (7), 1037-1041 (2005).</li><li>L&#248;bner, M., Walsted, A., Larsen, R., Bendtzen, K., Nielsen, C. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Enhancement+of+human+adaptive+immune+responses+by+administration+of+a+high-molecular-weight+polysaccharide+extract+from+the+cyanobacterium+Arthrospira+platensis.">Enhancement of human adaptive immune responses by administration of a high-molecular-weight polysaccharide extract from the cyanobacterium Arthrospira platensis.</a> Journal of Medicinal Food. 11 (2), 313-322 (2008).</li><li>Wang, H. B., Wu, S. J., Liu, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Preparation+of+polysaccharides+from+cyanobacteria+Nostoc+commune+and+their+antioxidant+activities.">Preparation of polysaccharides from cyanobacteria Nostoc commune and their antioxidant activities.</a> Carbohydrate Polymers. 99, 553-555 (2014).</li><li>Ozturk, S., Aslim, B., Suludere, Z., Tan, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Metal+removal+of+cyanobacterial+exopolysaccharides+by+uronic+acid+content+and+monosaccharide+composition.">Metal removal of cyanobacterial exopolysaccharides by uronic acid content and monosaccharide composition.</a> Carbohydrate Polymers. 101, 265-271 (2014).</li><li>Han, P. P., 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=Emulsifying,+flocculating,+and+physicochemical+properties+of+exopolysaccharide+produced+by+cyanobacterium+Nostoc+flagelliforme.">Emulsifying, flocculating, and physicochemical properties of exopolysaccharide produced by cyanobacterium Nostoc flagelliforme.</a> Applied Biochemistry and Biotechnology. 172 (1), 36-49 (2014).</li><li>Leite, J. P., 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=Cyanobacterium&#8208;Derived+Extracellular+Carbohydrate+Polymer+for+the+Controlled+Delivery+of+Functional+Proteins.">Cyanobacterium&#8208;Derived Extracellular Carbohydrate Polymer for the Controlled Delivery of Functional Proteins.</a> Macromolecular Bioscience. 17 (2), 1600206 (2017).</li><li>Estevinho, B. N., 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=Application+of+a+cyanobacterial+extracellular+polymeric+substance+in+the+microencapsulation+of+vitamin+B12.">Application of a cyanobacterial extracellular polymeric substance in the microencapsulation of vitamin B12.</a> Powder Technology. 343, 644-651 (2019).</li><li>Klock, J. H., Wieland, A., Seifert, R., Michaelis, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Extracellular+polymeric+substances+(EPS)+from+cyanobacterial+mats:+characterisation+and+isolation+method+optimisation.">Extracellular polymeric substances (EPS) from cyanobacterial mats: characterisation and isolation method optimisation.</a> Marine Biology. 152 (5), 1077-1085 (2007).</li><li>Delattre, C., Pierre, G., Laroche, C., Michaud, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Production,+extraction+and+characterization+of+microalgal+and+cyanobacterial+exopolysaccharides.">Production, extraction and characterization of microalgal and cyanobacterial exopolysaccharides.</a> Biotechnology Advances. 34 (7), 1159-1179 (2016).</li><li>Costa, T. R., 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=Secretion+systems+in+gram-negative+bacteria:+structural+and+mechanistic+insights.">Secretion systems in gram-negative bacteria: structural and mechanistic insights.</a> Nature Reviews Microbiology. 13 (6), 343-359 (2015).</li><li>Roier, S., Zingl, F. G., Cakar, F., Schild, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bacterial+outer+membrane+vesicle+biogenesis:+a+new+mechanism+and+its+implications.">Bacterial outer membrane vesicle biogenesis: a new mechanism and its implications.</a> Microbial Cell. 3 (6), 257-259 (2016).</li><li>Sergeyenko, T. V., Los, D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Identification+of+secreted+proteins+of+the+cyanobacterium+Synechocystis+sp.+strain+PCC+6803.">Identification of secreted proteins of the cyanobacterium Synechocystis sp. strain PCC 6803.</a> FEMS Microbiology Letters. 193 (2), 213-216 (2000).</li><li>Oliveira, P., 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=The+versatile+TolC-like+Slr1270+in+the+cyanobacterium+Synechocystis+sp.+PCC+6803.">The versatile TolC-like Slr1270 in the cyanobacterium Synechocystis sp. PCC 6803.</a> Environmental Microbiology. 18 (2), 486-502 (2016).</li><li>Flores, C., 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=The+alternative+sigma+factor+SigF+is+a+key+player+in+the+control+of+secretion+mechanisms+in+Synechocystis+sp.+PCC+6803.">The alternative sigma factor SigF is a key player in the control of secretion mechanisms in Synechocystis sp. PCC 6803.</a> Environmental Microbiology. 21 (1), 343-359 (2018).</li><li>Dubois, M., Gilles, K. A., Hamilton, J. K., Rebers, P. A., Smith, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Colorimetric+method+for+determination+of+sugars+and+related+substances.">Colorimetric method for determination of sugars and related substances.</a> Analytical Chemistry. 28 (3), 350-356 (1956).</li><li>Parikh, A., Madamwar, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Partial+characterization+of+extracellular+polysaccharides+from+cyanobacteria.">Partial characterization of extracellular polysaccharides from cyanobacteria.</a> Bioresource Technology. 97 (15), 1822-1827 (2006).</li><li>R&#252;hmann, B., Schmid, J., Sieber, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Methods+to+identify+the+unexplored+diversity+of+microbial+exopolysaccharides.">Methods to identify the unexplored diversity of microbial exopolysaccharides.</a> Frontiers in Microbiology. 6, 565 (2015).</li><li>Pathak, J., Rajneesh, R., Sonker, A. S., Kannaujiya, V. K., Sinha, R. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cyanobacterial+extracellular+polysaccharide+sheath+pigment,+scytonemin:+A+novel+multipurpose+pharmacophore.">Cyanobacterial extracellular polysaccharide sheath pigment, scytonemin: A novel multipurpose pharmacophore.</a> Marine Glycobiology. , 343-358 (2016).</li><li>Nguyen, A. T. 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=Performances+of+different+protocols+for+exocellular+polysaccharides+extraction+from+milk+acid+gels:+Application+to+yogurt.">Performances of different protocols for exocellular polysaccharides extraction from milk acid gels: Application to yogurt.</a> Food Chemistry. 239, 742-750 (2018).</li><li>Jamshidian, H., Shojaosadati, S. A., Mousavi, S. M., Soudi, M. R., Vilaplana, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Implications+of+recovery+procedures+on+structural+and+rheological+properties+of+schizophyllan+produced+from+date+syrup.">Implications of recovery procedures on structural and rheological properties of schizophyllan produced from date syrup.</a> International Journal of Biological Macromolecules. 105, 36-44 (2017).</li><li>Couto, N., Schooling, S. R., Dutcher, J. R., Barber, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Proteome+profiles+of+outer+membrane+vesicles+and+extracellular+matrix+of+Pseudomonas+aeruginosa+biofilms.">Proteome profiles of outer membrane vesicles and extracellular matrix of Pseudomonas aeruginosa biofilms.</a> Journal of Proteome Research. 14 (10), 4207-4222 (2015).</li></ol>

为了更好地了解细菌分泌机制和研究已释放的产品,证明在细胞外细菌环境中(如释放)中存在的生物分子的有效分离和分析具有极其重要的意义。碳水化合物聚合物和蛋白质)。
青基体细胞外碳水化合物聚合物极其复杂,主要是由于构成其成分1的不同单糖的数量和比例。用于分离这些聚合物物质的传统方法依赖于一个简单的概念,即这些富含糖的物质可溶于水溶?...

蓝藻被广泛认为是天然产品的丰富来源,具有广阔的生物技术/生物医学应用。因此,了解蓝藻分泌机制和优化提取/回收方法对于将蓝藻作为高效的微生物细胞工厂实施至关重要。
许多蓝藻菌株能够产生细胞外聚合物物质(EPS),主要由异聚糖形成,这些物质仍然与细胞表面相关或释放到介质1中。与其他细菌相比,这些释放的碳水化合物聚合物具有明显的特性,这使得它们适用于广泛的应用(例如,抗病毒药物2、免疫刺激3、抗氧化剂4、金属夹合5,乳化6,和药物输送剂7,8。分离这些聚合物的方法不仅有助于提高产量,而且有助于提高纯度,提高聚合物获得的具体物理特性。绝大多数这些方法分离聚合物依赖于沉淀策略从培养基,很容易完成由于聚合物的强烈电子性质9,10。此外,通过蒸发和/或冻干,可以迅速去除沉淀步骤中使用的溶剂。根据所预见的应用,在聚合物沉淀之后或之前可以耦合不同的步骤,以便定制最终产品,包括三氯乙酸 (TCA) 处理、过滤或尺寸排除色谱 (SEC)列净化10.
蓝藻也能够通过依赖膜输送器(经典)11或囊泡(非经典)12的通道分泌多种蛋白质。因此,分析蓝藻外蛋白体是了解/操纵蓝藻蛋白分泌机制和了解这些蛋白质的具体细胞外功能的重要工具。对外蛋白酶的可靠分离和分析需要细胞外环境的浓度,因为分泌蛋白质的丰度相对较低。此外,其他物理或化学步骤(例如离心、过滤或蛋白质沉淀)可以优化获得的外乳激素的质量,丰富蛋白质含量13,并避免污染物的存在(例如,色素、碳水化合物等)14,15或细胞内蛋白质在样本中的优势。然而,其中一些步骤也可能限制可以检测到的蛋白质集,从而导致有偏差的分析。
本作品描述了从蓝藻培养培养物中分离释放的碳水化合物聚合物和外丙酮的有效方案。这些协议可以很容易地适应研究的具体目标和用户需求,同时保持此处介绍的基本步骤。

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			<td height="21" style="height:21px;width:307px;">Dialysis membranes</td>
			<td style="width:191px;">Medicell Membranes Ltd&#160;</td>
			<td style="width:119px;">DTV.12000.07</td>
			<td style="width:777px;">Visking Tubing Size 7, Dia 23.8 mm, Width 39-41 mm 30m Roll&#160;</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:307px;">Ethanol 96%</td>
			<td style="width:191px;">AGA - &#193;lcool e G&#233;neros Alimentares, S.A.</td>
			<td style="width:119px;">4.000.02.02.00</td>
			<td>Fermentation ethyl alcohol 96% AGA</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">PES Filter 0.2 &#956;m</td>
			<td style="width:191px;">Fisher Scientific, Lda</td>
			<td style="width:119px;">15206869</td>
			<td>Syringe filter polystyrene 33MM 0.2&#181;M STR&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Amicon Ultra-15, Ultracel-3K</td>
			<td style="width:191px;">Merck Millipore Ltd.</td>
			<td style="width:119px;">UFC900324</td>
			<td>Centrifugal filters with a nominal molecular weight cut-off of 3 kDa</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Thermo Scientific&#160;Pierce BCA Protein Assay</td>
			<td style="width:191px;">Fisher Scientific, Lda</td>
			<td style="width:119px;">10741395</td>
			<td>Green-to-blue, precise, detergent-compatible assay reagent to measure total protein concentration</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Brillant Blue G Colloidal Concentrate&#160;</td>
			<td style="width:191px;">Sigma Aldrich Qu&#237;mica SL</td>
			<td style="width:119px;">B2025-1EA</td>
			<td>Coomassie blue&#160;</td>
		</tr>
	</tbody>
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looking outwards: isolation of cyanobacterial released carbohydrate polymers and proteins

蓝藻可以主动将多种生物分子分泌到细胞外环境中,如异质多糖和蛋白质。这些生物分子的鉴定和表征可以增进有关其分泌途径的知识,并有助于操纵它们。此外,其中一些生物分子在生物技术应用方面也很有趣。这里描述了两个协议,用于轻松和快速分离蓝藻释放的碳水化合物聚合物和蛋白质。释放的碳水化合物聚合物的分离方法基于使用有机溶剂在水溶液中多糖的传统沉淀技术。这种方法保留了聚合物的特性,同时避免了来自细胞碎片和培养基培养物的污染物的存在。在工艺结束时,冻干聚合物可供使用或定性,或可以进行进一步轮纯化,具体取决于最终预期用途。关于蓝藻外蛋白酶的分离,该技术基于通过离心和过滤去除主要污染物后无细胞介质的浓度。这种策略允许通过膜转运器或外膜囊泡可靠地分离通过细胞外环境的蛋白质。这些蛋白质随后可以使用标准质谱技术进行识别。这里提出的协议不仅适用于广泛的蓝藻,而且还适用于其他细菌菌株。此外,这些程序可以很容易地根据产品的最终用途、所需的纯度和细菌菌株进行定制。

图1描述了从蓝藻培养物中提取释放的碳水化合物聚合物的方法的原理图表示。图2显示了来自中度EPS生产商青霉素合成细胞的沉淀聚合物PCC 6803和高效EPS生产商Cyanothece sp.CCY 0110。 图3显示了不同程度污染的冻干聚合物,突出了离心步骤对最终产品纯度的重要性。图4描述了蓝藻外蛋白酶的隔离方法?...

Watch this Scientific Journal Video about Looking Outwards: Isolation of Cyanobacterial Released Carbohydrate Polymers and Proteins at JoVE.com

Looking Outwards: Isolation of Cyanobacterial Released Carbohydrate Polymers and Proteins

向外观察:分离氰化物释放的碳水化合物聚合物和蛋白质

这里描述了分离蓝藻释放的碳水化合物聚合物和分离其外蛋白酶的协议。这两种程序都体现了获得高纯度聚合物或蛋白质的关键步骤,可用于进一步分析或应用。它们也可以根据特定的用户需求轻松调整。

Cyanobacteria can actively secrete a wide range of biomolecules into the extracellular environment, such as heteropolysaccharides and proteins. The ...

我们开发的这些协议对于研究蓝藻及其释放的产品，即细胞外碳水化合物聚合物和蛋白质的分泌机制非常重要。所产生的知识将使我们能够定制这些产品的几个应用，即生物技术和生物医学应用。这些协议非常简单。它们可以根据生产有机体、用户需求和产品的最终应用进行定制。这些协议的主要优点是，它们可以很容易地适应其他生物系统，特别是其他细菌。这些协议很简单。然而，根据细菌菌株或所需的纯度程度，可能需要引入一些变化。首先，在标准条件下培养蓝藻菌株。使用标准协议衡量增长。接下来，将培养物转移到透析膜中，对至少10卷去化水进行透析，持续搅拌24小时。在4摄氏度下将培养在15，000倍g下离心15分钟。将上经剂转移到新小瓶中，然后丢弃颗粒。在4摄氏度下以20，000倍g再次离心15分钟，以去除污染物，如细胞壁碎片或脂质糖。离心后，将上经剂转移到玻璃烧杯上，然后丢弃颗粒。为了沉淀聚合物，在上总应物中加入两卷96%乙醇，并在一夜之间在4摄氏度下孵育悬浮液。对于少量或不可见的沉淀聚合物，在4摄氏度下以13，000倍g离心，25分钟。轻轻丢弃上一杯，将颗粒重新在一到两毫升的自洗脱化水中。将水悬浮液转移到小瓶中。对于可见、大量沉淀聚合物，将沉淀的聚合物与无菌金属钳收集到小瓶中。挤压聚合物，丢弃多余的乙醇。要执行冻干过程（也称为冻干），请将小瓶与沉淀的聚合物一起保持零下 80 摄氏度过夜。使聚合物干干至少48小时。然后，将干燥的聚合物储存在室温下干燥器中，直到进一步使用。要集中培养，首先在标准条件下培养蓝藻。使用标准程序监测蓝藻的生长。然后，在室温下以4000倍g离心，10分钟。将上一液转移到烧瓶中，然后丢弃细胞颗粒。接下来，通过 0.2 微米孔径过滤器过滤已脱芯介质。要浓缩介质约500次，请使用具有标称分子量的离心浓缩器，将4000倍g和15摄氏度的离心离心机离心，最大为1小时离心。离心后，用浓缩样品冲洗过滤装置样品储液罐壁，将含量转移到微离心管中。使用自洗培养培养介质对过滤装置样品储液罐执行额外的洗涤步骤，以确保最大限度的外蛋白回收。然后，将外蛋白样本储存在零下20摄氏度，直到进一步使用。必须大力添加乙醇，以优化聚合物的沉淀和产量。离心沉淀聚合物后，应小心地丢弃上先液，以避免可能松散地附着在烧瓶壁上的颗粒重新附着。如果进行手动过滤，则应温和避免破坏滤膜。一些沉淀的聚合物只有在离心后才能看到，主要是在烧瓶的墙壁上，例如来自Synechocystis的EPS。在其他情况下，如氰化聚合物，可以看到聚合物团在沉淀后漂浮在玻璃烧杯中。一些污染在聚合物冻干后很容易从宏观上检测，因为它们会改变聚合物色素沉着，通常是白色或浅棕色。橙色聚合物被含有这些色素的类胡萝卜素或结构污染。例如，绿色聚合物受到细胞碎片和叶绿素的污染。外蛋白组可能因细菌菌株而有显著差异。例如，参见来自单细胞蓝藻的外蛋白组和来自丝状菌株的外蛋白。外蛋白集中样品中多糖的大量数量会阻碍外蛋白组分析。例如，它可以延迟蛋白质分离和掩盖较少丰富的蛋白质。用户可以从成分、物理/化学性质、测试聚合物生物活性或改性聚合物成分方面对聚合物进行特征描述。对于蛋白质鉴定，可以进行凝胶中的蛋白质分离和质谱法。对于囊泡的分离，应进行外蛋白的超离心。在我们的小组中，这些协议已经为解开蓝藻分泌机制或细胞外碳水化合物聚合物在不同领域的应用铺平了道路，例如在生物修复或抗肿瘤或药物输送剂中。

Research

Education

JoVE Journal

JoVE Core

Cell Biology

Biochemistry

Unit 1

Biochemistry of the Cell

SCIENTISTS IN ACTION

High-throughput Screening of Carbohydrate-degrading Enzymes Using Novel Insoluble Chromogenic Substrate Assay Kits

使用新型不溶性生色底物检测试剂盒碳水化合物降解酶的高通量筛选

Carbohydrates active enzymes (CAZymes) have multiple roles in vivo and are widely used for industrial processing in the biofuel, textile, detergent, paper ...

科研

生物化学

Identification of Plant Ice-binding Proteins Through Assessment of Ice-recrystallization Inhibition and Isolation Using Ice-affinity Purification

植物冰结合蛋白通过冰再结晶抑制和隔离的评估使用冰亲和力纯化鉴定

Ice-binding proteins (IBPs) belong to a family of stress-induced proteins that are synthesized by certain organisms exposed to subzero temperatures. In ...

Isolation of Intermediate Filament Proteins from Multiple Mouse Tissues to Study Aging-associated Post-translational Modifications

从多个小鼠组织分离中间丝状蛋白以研究与老化相关的翻译后修饰

Intermediate filaments (IFs), together with actin filaments and microtubules, form the cytoskeleton &#8211; a critical structural element of every cell. ...

Biochemical and Structural Characterization of the Carbohydrate Transport Substrate-binding-protein SP0092

碳水化合物转运基质结合蛋白 SP0092 的生化和结构表征

Development of new antimicrobials and vaccines for Streptococcus pneumoniae (pneumococcus) are necessary to halt the rapid rise in multiple resistant ...

Analyzing Dynamic Protein Complexes Assembled On and Released From Biolayer Interferometry Biosensor Using Mass Spectrometry and Electron Microscopy

用质谱和电镜分析 Biolayer 干涉传感器上的动态蛋白复合物的组装和释放

In vivo, proteins are often part of large macromolecular complexes where binding specificity and dynamics ultimately dictate functional outputs. In this ...

Isolation of Glomeruli and In Vivo Labeling of Glomerular Cell Surface Proteins

Isolation of Glomeruli and In Vivo Labeling of Glomerular Cell Surface Proteins

肾小球的分离及肾小球细胞表面蛋白的活体标记

Proteinuria results from the disruption of the glomerular filter that is composed of the fenestrated endothelium, glomerular basement membrane, and ...

Fluorescent Silver Staining of Proteins in Polyacrylamide Gels

聚丙烯酰胺凝胶中蛋白质的荧光银染色

Silver staining is a colorimetric technique widely used to visualize protein bands in polyacrylamide gels following sodium dodecyl sulfate-polyacrylamide ...

Interfacial Molecular-level Structures of Polymers and Biomacromolecules Revealed via Sum Frequency Generation Vibrational Spectroscopy

通过相频生成振动光谱揭示聚合物和生物大分子的界面分子级结构

As a second-order nonlinear optical spectroscopy, sum frequency generation (SFG) vibrational spectroscopy has widely been used in investigating various ...

Localization of SUMO-modified Proteins Using Fluorescent Sumo-trapping Proteins

荧光苏门答腊蛋白对 sumo 修饰蛋白的定位

Here we are presenting a novel method to study the sumoylation of proteins and their sub-cellular localization in mammalian cells and nematode oocytes. ...

Preparation of High-Quality Fermented Fish Product

优质发酵鱼产品的制备

This protocol provides a method for preparation of industrialized fermented fish product with sturgeon (Aquilaria sinensis) meat product. The procedures ...