叶蛋白提取方法的比较及发酵酸法的应用前景

概述了叶蛋白提取的几种方法,其中有水溶液提取法、加热法(60℃加热、90℃加热)、酸碱法、纯蛋白质沉淀法、盐析法、凝聚剂沉淀法、超滤法、电浓缩法、反胶团相转移法、有机溶剂沉淀法、发酵酸法等。将发酵酸法和其他的提取方法进行了比较,重点讲述了发酵法提取叶蛋白的优点,并指出叶蛋白在今后开发利用中的良好前景。     

紫云英叶蛋白提取工艺研究

本文采用正交试验对紫云英叶蛋白提取工艺进行了研究,结果表明:蛋白质萃取工艺较优条件为:萃取剂与紫云英鲜叶料液比为5:1,萃取温度为20℃,萃取pH值7.0,萃取剂中盐浓度为0.5％;绿色蛋白质在50℃时沉淀,白色蛋白质在pH=5.0时沉淀。采用此工艺提取得率为25.4％,每吨紫云英鲜叶可得粗蛋白0.04吨,所得粗蛋白含真蛋白50％左右。     

芽胞杆菌菌体蛋白提取方法的比较

目的：比较不同破碎方法下3种芽胞杆菌菌体蛋白的提取效果。方法：以菊酯类农药降解菌巨大芽胞杆菌、球形芽胞杆菌、弯曲芽胞杆菌为研究对象,以裂解液为提取介质,分别采用超声波破碎法、水煮法和机械破碎法提取芽胞杆菌蛋白,并通过革兰染色电镜观察菌株破碎程度,用SDS-PAGE和Bradford法测定蛋白浓度。结果：革兰染色电镜观察发现玻璃珠机械破碎法对3种芽孢杆菌的细胞壁破碎程度最明显,其次为超声波破碎法,水煮法破碎效果不明显。SDS-PAGE和Bradford法测定结果表明,巨大芽胞杆菌、球形芽胞杆菌、弯曲芽胞杆菌经玻璃珠机械破碎后,提取的蛋白图谱条带清晰,丰度高,重复性好,含量分别为20.247、19.902和18.893 mg/mL;经超声波破碎提取的蛋白图谱条带较清晰,丰度一般,重复性不好,含量分别为10.572、9.438和10.424 mg/mL;经沸水浴破碎提取的蛋白图谱条带模糊,丰度低,重复性差,含量分别为1.366、1.119和1.136 mg/mL。结论：玻璃珠机械破碎法是破碎3种芽胞杆菌的最优方法,破碎后提取的蛋白含量高,条带清晰,重复性好。     

盾叶薯蓣中薯蓣皂甙元不同提取方法的比较

为提高盾叶薯蓣(Dioscorea zingiberensis C.H.Wright)中薯蓣皂甙元得率,在实验室条件下,以薯蓣皂甙元含量为评价指标,采用4因素4水平的正交试验,用高效液相色谱法对16种提取方法所得的薯蓣皂甙元含量进行了比较分析.结果表明,硫酸的浓度对薯蓣皂甙元含量有极显著影响,在实验室条件下,可采用10 g样品加2.0mol·L-1硫酸200 mL水解4 h,水解物用石油醚回流提取5 h,能快速准确提取盾叶薯蓣中的薯蓣皂甙元.     

银杏叶黄酮提取方法比较

比较不同溶剂提取银杏（ＧｉｎｋｇｏｂｉｌｏｂａＬ．）叶黄酮类化合物的提取效率,从成本效益角度考虑,以７０％乙醇作为提取溶剂更为有利。在分级沉淀中,黄酮含量与蛋白质含量呈正相关,在乙醇提取液中黄酮和蛋白质含量最高;蛋白质的存在有助于提高黄酮的溶解度。乙醇提取液用饱和（ＮＨ４）２ＳＯ４浓缩两次,可使醇相中的黄酮沉淀析出。根据试验结果,提出了银杏叶黄酮的优化提取方法。     

蚜虫全蛋白提取方法的比较研究

为建立适于SDS-PAGE分析的蚜虫蛋白质样品制备平台,以便为蚜虫蛋白质的双向电泳分析奠定基础,本研究比较了TCA/丙酮沉淀、PEG提取、饱和酚抽提和直接裂解4种蛋白质提取方法.结果表明:不同样品制备方法的蛋白提取率有显著的差异,其中直接裂解法的提取率最高,为17.43 mg/g;其次是饱和酚抽提法,提取率为12.30 mg/g;而PEG制备法提取率最低,只有7.96 mg/g.利用SDS-PAGE电泳对不同的蛋白质样品进行了分析,发现在凝胶图谱上显现的条带也有明显的差异,其中饱和酚抽提法显现的条带数最多,为36条,且从14.4 kDa～116.0 kDa范围有广泛分布,条带清晰;PEG提取法条带数为30条,一些蛋白条带丢失或不明显;TCA/丙酮沉淀法的蛋白条带集中分布在25.0 kDa～67.0 kDa区域;直接裂解法条带数仅为24条,且小分子量的条带可辩率很低.通过以上结果可以得出,饱和酚抽提法最适用于蚜虫全蛋白样品的制备.     

塔玛亚历山大藻双向电泳蛋白的三种提取方法比较

【目的】比较3种蛋白质提取方法,找到适用于塔玛亚历山大藻蛋白的最佳的提取方法,为后续用双向电泳(2-DE)技术研究不同条件下塔玛亚历山大藻蛋白的差异表达奠定基础。【方法】以塔玛亚历山大藻为研究对象,运用Tris-HCl提取法、TCA沉淀法和lysis buffer提取法分别提取塔玛亚历山大藻蛋白,并通过双向电泳技术,对这3种方法进行了比较分析,筛选出最适于塔玛亚历山大藻的蛋白提取方法。并运用以上得出的方法,以不加杀藻物质的无菌塔玛亚历山大藻为对照,比较分析了塔玛亚历山大藻在加入杀藻物质后的蛋白差异表达状况。【结果】在这3种方法中,lysis buffer提取法得到的蛋白溶解性好,进行双向电泳时,可得到干净的背景、清晰的蛋白点,并且蛋白点的数目较多,酸性蛋白、碱性蛋白、大分子量和小分子量的蛋白均有提出来,蛋白点在胶面上分布均匀。用这种方法初步分析了加入杀藻物质后塔玛亚历山大藻蛋白的差异表达情况,并鉴定出14个与塔玛亚历山大藻生理活动密切相关的蛋白质。【结论】lysis buffer提取法获得了最多的蛋白点,双向电泳图谱清晰,适于用来提取塔玛亚历山大藻蛋白。     

纳米器件对叶蛋白提取的影响

从植物中提取蛋白质包括机械破碎和离心等一系列步骤,蛋白质在提取液中的溶解度直接影响蛋白提取率和产量.结果表明,用纳米器件处理过的提取液能显著提高叶蛋白的提取率,提升幅度为10％-30％.利用正交试验研究不同提取条件下纳米器件对蛋白可溶性的影响,结果显示浸泡时长(纳米器件浸泡在提取液中的时长)是最关键的已测因素.此外,植物样品的破碎程度也是影响叶蛋白提取率的关键因素之一.     

Economic comparison of multiple techniques for recovering leaf protein in biomass processing

Leaf protein concentrates (LPC) can be used as a valuable co-product to cellulosic biofuel production and can also mitigate the food versus fuel controversy. Two major approaches have been considered for LPC production: a well-characterized mechanical pressing method and a less studied method involving aqueous extraction with recovery using ultrafiltration. Experimental results with switchgrass extracts show low protein recovery after filtration, particularly if protein is recovered after cellulose hydrolysis. Economic modeling suggests that aqueous extraction costs less than mechanical pressing, but due to lower protein yields and lower quality, overall profit is higher for mechanical pressing versus aqueous extraction ($26/Mg feedstock vs. $14/Mg). If modest improvements can be made in extraction yields, filtration recovery, and protein quality, then the profitability of the aqueous extraction approach can be increased to $37/Mg feedstock. This study suggests that aqueous extraction is a viable alternative for LPC co-production in a biorefinery if key improvements can be made in the process.     

黄粉虫有效物质的综合提取及提取方法的比较

比较了用石油醚和用环己烷抽提黄粉虫TenebriomolitorL .虫粉中脂肪的提取率 ,以及用碱法和用酶法提取去脂虫渣中蛋白质的提取率。并从 2种去脂、去蛋白质虫渣中和虫蜕中提取壳聚糖。用环己烷和酶法提取为最佳组合 ,可综合提取出占幼虫干重 5 0 .5 %的有效物质。同时 ,讨论了碱法和酶法提取率出现差异的原因。     

深山含笑叶片总酚超声波提取工艺的优化

采用单因素实验方法研究了超声波辅助提取过程中乙醇浓度、料液比、提取温度和提取时间对深山含笑（Michelia maudiae Dunn）叶片总酚提取率的影响,并采用正交实验方法确定了最佳提取工艺条件。结果表明,深山含笑叶片总酚超声波辅助提取的最佳提取工艺为：按1：30（质量-体积比）的料液比加入体积分数70％的乙醇,于65℃条件下用超声波辅助提取30min。采用最佳的超声波提取工艺,深山含笑叶片的总酚提取率可达到11．41％。定性分析结果显示,深山含笑叶片的总酚提取物具有典型的酚类化合物特性,并显示出鞣质类成分、黄酮类成分和香豆素类成分的定性反应特征。     

不同工艺提取老鹰茶总黄酮含量的比较

比较不同工艺提取富集的老鹰茶中总黄酮含量,分别采用传统乙醇提取-正丁醇萃取法和水提取-大孔树脂法提取老鹰茶总黄酮,并用紫外分光光度法,以芦丁为标准品,270nm下测定总黄酮含量。水提取-大孔树脂法富集所得固体中黄酮含量较高,可达55.86%,回收率为105.6%,RSD=1.64%。水提取-大孔树脂富集的工艺操作简单,所得固体黄酮含量高,更适于老鹰茶总黄酮的提取,对进一步的工业化生产开发有较好的参考价值。     

Evaluation of protein extraction methods to improve meta-proteomics analysis of treated wastewater biofilms

Metaproteomics can be used to study functionally active biofilm-based bacterial populations in reclaimed water distribution systems, which in turn result in bacterial regrowth that impacts the water quality. However, existing protein extraction methods have differences in their protein recovery and have not been evaluated for their efficacies in reclaimed water biofilm samples. In this study, we first evaluated six different protein extraction methods with diverse chemical and physical properties on a mixture of bacterial cell culture. Based on a weighting scores-based evaluation, the extraction protocols in order of decreasing performance are listed as B-PER > RIPA > PreOmics > SDS > AllPrep > Urea. The highest four optimal methods on cell culture were further tested against treated wastewater non-chlorinated and chlorinated effluent biofilms. In terms of protein yield, our findings showed that RIPA performed the best; however, the highest number of proteins were extracted from SDS and PreOmics. Furthermore, SDS and PreOmics worked best to rupture gram-positive and gram-negative bacterial cell walls. Considering the five evaluation factors, PreOmics obtained highest weighted score, indicating its potential effectiveness in extracting proteins from biofilms. This study provides the first insight into evaluating protein extraction methods to facilitate metaproteomics for complex reclaimed water matrices.     

观察植物叶迹、叶隙立体结构的实验方法

通过实验研究提出了观察植物叶迹、叶隙立体结构的方法。对观察节处维管组织的变化, 有重要意义。     

Effect of leaf surface extraction on palatability of romaine lettuce to Diabrotica balteata

Abstract The leaf surface of a plant, especially its chemical components, constitutes the first line of resistance to herbivores and other pests. Our previous research indicated that ‘Valmaine’ (Val) romaine lettuce, Lactuca sativa L., was highly resistant to feeding by adult banded cucumber beetle, Diabrotica balteata LeConte, while ‘Tall Guzmaine’ (TG) was highly susceptible. We investigated the leaf surface chemistry of these two cultivars for its possible role in their resistance to D. balteata. Three solvents with different polarity (hexane, methylene chloride, and methanol) were tested to remove leaf surface chemicals, but only methylene chloride and methanol extracts were used in feeding bioassays. Adult D. balteata consumed much more of the leaf tissue of Val and TG when their surface chemicals were removed with methylene chloride, but not methanol, compared to nonextracted leaf tissue, leading us to hypothesize that methylene‐chloride extractable leaf surface chemicals may have a role in the expression of lettuce resistance. However, leaf surface chemicals extracted from Val with methylene chloride were not a deterrent to adult D. balteata when applied to palatable lima bean leaf surfaces at various concentrations in dual‐choice tests. Furthermore, the application of surface extracts from TG did not stimulate beetle feeding in similar choice tests. In a no‐choice feeding test, there was no significant difference in leaf area consumption on lima bean leaves sprayed with extracts of Val or TG. These results suggest that leaf surface chemicals in romaine lettuce do not explain the resistance of Val to adult D. balteata, and that factors inside the leaf may play a role in resistance. We discuss the possibility that the solvent may have increased the palatability of lettuce leaves to D. balteata by causing enzymatic browning and cellular damage, which is likely to have degraded internal feeding deterrents and impaired the plant's ability to emit latex.     

Varietal differences of leaf protein profiles in mulberry

Varietal differences in the leaf protein profiles of 17 diploid (2n = 28) varieties of mulberry (Morus spp.) were demonstrated by 2D-gel electrophoresis and silver staining; similarity was statistically evaluated by means of principal component analysis. A close correlation was found between inter-varietal similarity, as estimated by the electrophoretic banding patterns of certain enzymes, and the leaf protein profiles obtained by 2D-gel electrophoresis. Little apparent varietal difference in the amount of ribulose-1,5-bisphosphate carboxylase was detected immunoelectrophoretically in the leaves. Varietal differences in crude protein content were obtained with leaves and this was discussed in relation to the observed 2D-gel electrophoretic patterns.     

30种植物叶蛋白中氨基酸组成及含量的测定与营养价值评价

评价植物叶蛋白的营养价值,既要考虑“量”,看植物叶蛋白含量的高低,也要考虑“质”,看必需氨基酸的含量及配比[1]。作者取叶蛋白含量50%以上的30种植物进行氨基酸含量测定[2],并进行营养价值评价。1　方　　法应用日立83550型氨基酸自动分析仪进行氨基酸含量的测定,并与?..