冷榨菜籽蛋白的碱性电解水提取工艺及其结构性能表征

为探寻一种油料蛋白提取方法,该研究以双低冷榨菜籽饼为原料,碱性电解水为溶剂,通过单因素与响应面试验确定了碱性电解水提取冷榨菜籽饼分离蛋白（Alkaline electrolyzed water extracted Rapeseed Protein,ARP）的优化条件,并与相同条件下碱溶酸沉法提取菜籽分离蛋白（Ultrapure water extracted Rapeseed Protein,URP）的结果进行对比。结果表明: 提取工艺各环节因素对菜籽蛋白提取率影响的主次顺序分别为：碱性电解水pH值、温度、料液比、时间;提取优化工艺条件为温度45 ℃、碱性电解水pH值11.5、料液比1:10 g/mL和提取时间60 min,蛋白质提取率为59.34%。与相同条件下碱溶酸沉法相比,碱性电解水提取的菜籽分离蛋白游离巯基和二硫键含量高、表面疏水性强、粒径小、ζ电位绝对值大,同时其提取率、溶解性、持油性、乳化性和起泡性均显著得到改善（P＜0.05）;此外,ARP内源荧光光谱强度更高,二级结构更有序。由此可知,该研究结果为碱性电解水提取冷榨菜籽饼蛋白提供参考,不仅提高了提取率,而且最大程度地减少了对菜籽蛋白结构和功能特性的破坏。     

高粱外种皮中原花青素的提取工艺及其组分鉴定

本研究旨在提取黑龙江产高粱外种皮中的原花青素并鉴定其组成.通过正交试验对提取工艺进行优化,采用柱层析、葡聚糖(Sephadex LH-20)凝胶色谱进行纯化和分离,纯化和分离的原花青素产物用ESI-MS鉴定其组成.在影响提取率的4个因素温度、料液比、时间和pH值中, 温度对提取率的影响最为显著,料液比也有一定影响.研究得到最佳提取工艺为:温度80℃,时间1.5 h,pH值为4,料液比为1∶20;通过ESI-MS鉴定出了原花青素的组成成分的分子量,黑龙江产高粱外种皮中的原花青素以聚合度小于5的低聚体为主.     

碱酶两步法制备大米蛋白的研究

该文采用碱法和酶法两步加工碎米，制备得到纯度较高的大米蛋白。通过研究米粉粒度、pH值、温度、水料比和时间对提取率的影响，确定碱法制备大米蛋白的较佳工艺条件为：米粉粒度80目，pH 11.0，温度50℃，水料比8，时间120 min。采用α-淀粉酶对大米蛋白进行纯化，酶解条件为：加酶量60 U/g，pH 6.0，温度50℃，时间30 min。在上述工艺条件下制备大米蛋白，提取率为73.22%，纯度为88.75%。     

pH和肥料对茶园土壤基础呼吸的影响

对茶园土壤添加不同用量石灰处理形成三种不同pH值的土壤,采用室内培养法,通过添加菜籽饼和硫酸铵,研究了pH及碳氮互作对茶园土壤基础呼吸速率的影响。结果表明:在培养的28天中,土壤基础呼吸速率表现出先高后低的趋势,不同pH值的茶园土壤以pH 4.05的土壤最高,显著高于其它2个土壤,pH 4.22土壤次之,pH 3.60土壤最低;添加菜籽饼可以显著增加土壤的呼吸速率,加氮对土壤基础呼吸速率的影响不明显;同时添加菜籽饼和氮肥在pH 4.22的土壤呼吸有协同增效作用,但在pH 3.60和pH 4.05的土壤不明显。试验还表明,土壤基础呼吸与土壤微生物量碳呈极显著正相关,但与土壤水溶性有机碳含量相关性不明显。     

水相酶解法提取菜籽油与菜籽蛋白工艺的优化

为提高菜籽蛋白的利用率，采用水相酶解法提取菜籽油与菜籽蛋白，以超滤法分离菜籽蛋白，重点对酶处理的工艺参数进行了研究。试验对各种酶的作用效果进行了比较，确定采用纤维素酶、果胶酶按活力比3∶1配方的复合酶处理效果较好；由优化试验得出酶处理的工艺参数为：固液比1∶5，酶用量30 U/g，酶处理时间100 min；在最佳工艺条件下获得高质量的菜籽油与菜籽蛋白，油与蛋白质得率分别为92.6%、82.3%。     

鲤鱼精蛋白的提取与抗菌稳定性研究

该文研究了采用超声波技术提取鲤鱼抗菌精蛋白的工艺条件和抗菌稳定性。实验结果表明：采用超声波技术提取鱼精蛋白硫酸用量少,提取时间短。用1.0A强度的超声波和5%的硫酸溶液提取鱼精蛋白,提取时间20 min,能得到较高活性的鲤鱼抗菌精蛋白。并进一步研究了pH值、温度、金属离子和蛋白酶对鲤鱼精蛋白抗菌活性的影响。     

酶法有限水解含油菜籽蛋白的机理及动力学

为探索油菜籽水相酶解法萃取菜籽油的机理,以Alcalase蛋白酶在温度50℃、pH8.0条件下,对以水剂法从菜籽中提取的含油菜籽蛋白进行酶解处理.试验研究结果表明:含油菜籽蛋白乳液的固形物颗粒度对酶解反应速率有明显影响,总的趋势是颗粒度越小,酶促反应速率越大;酶催化水解速率随水解进程呈指数下降,在反应过程中过高的底物浓度会抑制酶的失活.在此基础上由试验数据推导出描述催化水解含油菜籽蛋白的动力学方程,由此通过控制酶与底物浓度之比、反应时间,可以控制水解作用的程度,从而指导和优化菜籽水相酶解法提取菜籽油的工艺.     

菜籽饼堆肥对水稻土壤Cd有效性及Cd在水稻全生育期转运与累积的影响

为研究菜籽饼堆肥对土壤Cd有效性和在水稻体内迁移转运与累积的影响,在Cd污染(Cd=0.72 mg/kg)土壤中施用不同添加量(0.75%,1.5%,3.0%)的菜籽饼堆肥,以未添加菜籽饼堆肥为对照(CK),并进行水稻盆栽种植试验。结果表明:(1)菜籽饼堆肥进入稻田土壤后会显著降低土壤中TCLP提取态Cd含量,在熟化期施用0.75%～3.0%的菜籽饼堆肥,与对照相比土壤TCLP提取态Cd含量下降了45.1%～68.7%。但水稻的种植会影响菜籽饼堆肥对土壤中TCLP提取态Cd含量的降低效果,使其含量随着水稻生育期的延长逐渐上升,但仍低于同时期的对照土壤。(2)施用菜籽饼堆肥能显著提高水稻产量,但同时也增加水稻糙米中Cd含量。与对照相比,施用0.75%～3.0%的菜籽饼堆肥,水稻糙米中Cd含量为0.04～0.14 mg/kg,低于国家食品中污染物限量标准(GB 2762—2012, Cd<0.2 mg/kg)。同时,每株水稻产量分别增加3.6～4.3 g/株,约为1 620～1 935 kg/hm^2。(3)施用菜籽饼堆肥会提高Cd在水稻体内的转运能力,同时显著提高水稻成熟期各部位Cd累积量,特别是地上部分。总体来说,施用菜籽饼堆肥增加水稻糙米中Cd含量,但依然保持在较低水平,满足中轻度Cd污染地区水稻的安全生产。但在Cd污染程度更高或者土壤Cd活性更强的土壤中施用菜籽饼堆肥,种植水稻糙米Cd含量可能高于国家食品中污染物限量标准。因此,在保证稻米安全的前提下对Cd污染稻田应该谨慎施用菜籽饼堆肥。     

菜籽蛋白加工废液中多酚和多糖同步提取工艺优化

为开发利用菜籽蛋白加工废液中的生理活性物质,该研究在单因素试验基础上,采用Box-Behnken响应面试验设计法,对菜籽蛋白加工废液中多酚和多糖提取工艺条件进行优化,同时探究两种物质的体外抗氧化活性。结果表明,影响菜籽蛋白加工废液中多酚和多糖得率的因素大小顺序为:乙醇体积分数浸提温度浸提时间,最佳提取工艺为:浸提温度60℃、乙醇体积分数65%、浸提时间31 min,在此条件下多酚得率为2.19%,多糖得率为8.14%;多酚提取物对DPPH·具有较强清除能力,其半抑制质量浓度为0.20 mg/mL,多糖提取物对DPPH·和·OH均具有较强的清除能力,其半抑制质量浓度分别为1.45、2.38 mg/mL;高效液相色谱法初步检测表明,菜籽蛋白加工废液中含有香豆酸、丁香酸、对香豆酸、芥子酸和苯甲酸。研究结果为菜籽蛋白加工废液的再利用提供参考。     

响应面优化花生叶可溶性蛋白提取工艺及抗氧化活性分析

为确定花生叶可溶性蛋白提取最佳工艺,本试验以花生叶为材料,利用碱溶酸沉法提取花生叶可溶性蛋白,分析浸提pH值、料液比及提取时间对蛋白提取率的影响;在单因素试验的基础上,采用Box-Behnken响应面分析法(RSM)优化花生叶可溶性蛋白提取条件,并对所得蛋白进行抗氧化活性分析。结果表明,各影响因素主次顺序为浸提pH值>提取时间>料液比,最佳提取条件为料液比1∶20(w/v)、提取时间40 min、浸提pH值 9.0,以此条件建立的响应面试验回归模型拟合性好(R²=0.992 7),花生叶可溶性蛋白理论提取率为52.8%,实际提取率为54.2%,纯度为63.8%。花生叶可溶性蛋白具有较强的DPPH自由基、超氧阴离子自由基清除能力,以及较好的还原能力。本研究结果为花生叶的开发利用及花生产业链的延长提供了一定的数据支持。     

Comparison of protein chemical and physicochemical properties of rapeseed cruciferin with those of soybean glycinin

Rapeseeds contain cruciferin (11S globulin), napin (2S albumin), and oleosin (oil body protein) as major seed proteins. The effects of oil expression and drying conditions on the extraction of these proteins from rapeseed meal were examined. The conditions strongly affected the extraction of oleosin and only weakly affected the extraction of cruciferin and napin. The protein chemical and physicochemical properties of cruciferin, the major protein present, were compared with those of glycinin (soybean 11S globulin) under various conditions. In general, cruciferin exhibited higher surface hydrophobicity, lower thermal stability, and lower and higher solubility at mu= 0.5 and mu = 0.08, respectively, than did glycinin. At the pHs (6.0, 7.6, and 9.0) and ionic strengths (mu= 0.08 and 0.5) examined, the emulsifying ability of cruciferin was worse than that of glycinin, except at mu= 0.08 and pH 7.6. The emulsifying abilities of cruciferin and glycinin did not correlate with thermal stability and surface hydrophobicity. Higher protein concentration, higher heating temperature, higher pH, and lower ionic strength were observed to produce harder gels from cruciferin. Gel hardness partly correlated with the structural stability of cruciferin.     

超声辅助提取辣椒籽蛋白工艺优化

以辣椒籽为原料,采用超声辅助法提取辣椒籽蛋白并利用响应面优化法提取工艺,在单因素试验的基础上,选取p H值,提取时间,超声功率,料液比4个因素进行响应面试验,根据所得试验结果确定最佳提取条件为:p H值11,提取时间13.31 min,超声功率336.21 W,料液比1∶35.85,在此条件下蛋白的预计提取量为5.90 g/(100 g)。利用Design expert软件对影响辣椒籽蛋白提取量的主要因素及其相互作用进行探讨,结果由大到小依次为:p H值料液比提取时间超声功率。与传统提取方法相比,超声辅助法使得蛋白提取量增加了0.81 g/(100 g)(占传统方法提取量的15.46%),蛋白纯度提高了5.47%。     

蛋白原料及其混合粉料理化性质对颗粒饲料加工质量的影响

该研究旨在探究豆粕、棉粕、菜粕、酒糟蛋白(Distillers Dried Grains with Solubles,DDGS)、乙醇梭菌蛋白5种蛋白原料及其混合粉料的营养指标和理化性质的差异,确定影响颗粒饲料质量和制粒能耗的关键指标,对5种蛋白原料的制粒效果进行综合评价。以豆粕为对照组,仅改变蛋白原料,采用相同的加工参数制备颗粒饲料,比较不同蛋白原料的制粒效果,进行主成分分析及偏最小二乘回归分析(Partial Least Squares Regression,PLS)。结果表明:在原料营养指标和理化特性方面,乙醇梭菌蛋白具有高蛋白含量、高蛋白溶解度、低脂肪、低纤维的特点,棉粕具有高纤维的特点,菜粕具有高纤维和低蛋白溶解度的特点,DDGS具有低蛋白和高脂肪的特点。蛋白原料吸水性强弱排列顺序为乙醇梭菌蛋白、豆粕、棉粕、菜粕、DDGS,水溶性与之相反。乙醇梭菌蛋白组和棉粕组的制粒能耗较高,豆粕组的制粒能耗最低;棉粕组和乙醇梭菌蛋白组的修正耐久性(Modified Pellet Durability Index,MPDI)较高分别为92.72%和90.57%,菜粕组的MPDI最低为79.68%;乙醇梭菌蛋白组的硬度最高为130.95N,DDGS组的硬度最低为74.26N;乙醇梭菌蛋白组的糊化度最高为45.56%,DDGS组的糊化度最低为31.36%。通过偏最小二乘回归模型得到,蛋白含量、蛋白溶解度和吸水性的增加会提高颗粒饲料硬度、PDI和MPDI;粗纤维含量、蛋白溶解度和吸水性的增加会增加制粒能耗。综合分析5种蛋白原料制粒特性,由高到低排序为乙醇梭菌蛋白、棉粕、豆粕、菜粕、DDGS。研究结果为实际生产颗粒饲料时蛋白原料的选择提供参考依据。     

Impact of isolation method on the antioxidant activity of rapeseed meal phenolics

Rapeseed meal is the byproduct of the rapeseed deoiling process. Among oilseed plants, rapeseed contains the greatest amount of phenolic compounds. In this study, the rapeseed phenolics were isolated with aqueous methanol, aqueous ethanol, hot water, and enzymatically with ferulic acid esterase. These isolates were tested for radical scavenging and for liposome and low-density lipoprotein (LDL) model systems. The radical scavenging activities of all isolates were >60% at a concentration of 1.5 mg/mL. In the liposome model system the formation of hexanal was inhibited by all rapeseed meal isolates by >90% and the formation of conjugated diene hydroperoxides by >80% (8.4 microg/mL concentration). All rapeseed meal isolates also inhibited oxidation of LDL particles by >90% inhibition (4.2 microg/mL concentration). Isolation of rapeseed meal phenolics with either water or enzyme is a very suitable method devoid of organic solvents. Thus, rapeseed meal phenolics constitute an interesting source for food and cosmetic applications with antioxidant effect.     

白菜型冬油菜‘陇油7号’叶片响应低温胁迫的差异蛋白鉴定与分析

为了从蛋白质组学的角度探讨超强抗寒品种白菜型冬油菜‘陇油7号’的抗寒机理,本研究采用TCA(三氯乙酸)-丙酮沉淀法,提取低温胁迫(4℃,7 d)前后的叶片总蛋白,对蛋白提取方法、IPG(固定pH梯度)胶条种类等环节进行了优化,运用双向电泳和质谱分析技术,鉴定了低温胁迫下‘陇油7号’5叶期叶片总蛋白质组分的表达差异模式。结果表明:改进后的蛋白质提取液(含DTT,二硫苏糖醇)和PVPP(聚乙烯吡咯烷酮)得到的蛋白质平均浓度较改进前高3.42μg·μL-1,除盐时间较改进前短1.14 h;同时,含蛋白酶抑制剂苯甲基磺酰氟(PMSF)的蛋白提取液获得的蛋白质种类丰富,凝胶图谱中可检测到661个蛋白点,较改进前可测蛋白点数(587)提高11.2。采用17 cm p H 4~7的IPG胶条的电泳能更好地分离蛋白,得到重复性好、分辨率高的蛋白质组图谱。利用PDQuest 8.0软件分析比较了超强抗寒品种‘陇油7号’低温胁迫前后的蛋白组表达谱,发现低温胁迫前后共有15个质变的蛋白质点,推测这些差异蛋白点可能与低温胁迫的响应有关。进一步对质变的蛋白质点进行了质谱分析,鉴定出11个与低温胁迫相关的蛋白质点,这些蛋白包括光合作用相关的蛋白、糖代谢相关的蛋白、物质运输相关的蛋白和逆境响应相关的蛋白。而且,低温胁迫处理前后,‘陇油7号’叶片蛋白质的表达水平存在明显差异,这些差异蛋白可能在冬油菜抗寒响应中发挥重要作用。     

菜籽粕植酸提取和分离蛋白的制备

植酸和蛋白是菜籽粕中2种极具经济价值的成份。为提高菜籽粕的综合利用效果,该文以双低冷榨菜籽粕为原料,采用醋酸溶液提取植酸,在膜分离精制植酸粗提液过程中同时回收蛋白;再对植酸提取后的残余物进行蛋白分离,超滤纯化后获得高纯度的蛋白成品。响应面优化的植酸最适提取条件为：醋酸质量分数0.7%,提取温度48℃,液料比10 mL/g,提取时间1.6 h,该条件下植酸得率为1.865%。植酸粗提液中回收出的蛋白和损失植酸分别占菜籽粕的3.63%和0.395%。超滤精制的分离蛋白可达到70%～90%不同纯度的要求,蛋白中多酚含量显著减少,且植酸与硫苷未检出。     

Determination of multiresidues in rapeseed, rapeseed oil, and rapeseed meal by acetonitrile extraction, low-temperature cleanup, and detection by liquid chromatography with tandem mass spectrometry

A multiresidue method for determining pesticides in rapeseed, rapeseed oil, and rapeseed meal by use of liquid chromatography-tandem mass spectrometry is developed. Samples were extracted with acetonitrile or acidified acetonitrile and cleaned up by a 12 h freezing step. The recovery data were obtained by spiking blank samples at three concentration levels. The recoveries of 27 selected pesticides in rapeseed, rapeseed oil, and rapeseed meal were in the range of 70-118%, at the concentration level of 10 μg kg(-1), with intraday and interday precisions of lower than 22 and 27%, respectively. Linearity was studied between 2 and 500 μg L(-1) with determination coefficients (R(2)) of higher than 0.98 for all compounds in the three matrices. The limits of quantitation (LOQs) of pesticides in rapeseed, rapeseed oil, and rapeseed meal ranged from 0.3 to 18 μg kg(-1). The n-octanol-water partition coefficient showed more influence than water solubility in extracting pesticides by acetonitrile from matrices of high fat content. This method was successfully applied for routine analysis in commercial products.     

Determination of virginiamycin in feeds.

Virginiamycin was extracted from the feed by ethanol-pH 2.5 phosphate buffer (1 + 1). The pH during extraction was adjusted (when necessary) to between 4 and 5. Sample dilutions and the standard dose response line were prepared to contain ethanol pH 6 phosphate buffer (2 + 8), and the test organism was Sarcina lutea. Three feeds (a poultry ration, a swine finishing ration, and a swine starter ration) showed virginiamycin recovery of 88.8--108.9% when standard solutions were added at concentrations of 4.54--90.8 g/ton. The coefficient of variation (4--20%) was larger for low potency feeds (10 g/ton) compared to the higher feeds (100 g/ton). Similarly, excellent recovery was obtained when the swine starter feed was fortified by a commercial premix. Amprolium, roxarsone, and monensin can be present at 20 times the concentration of virginiamycin with little or no interference in the antibiotic determination. Lasalocid at 10 times the concentration of virginiamycin caused a slightly positive bias (recovery, 107.4%).     

Physicochemical and flavor characteristics of flavoring agent from mungbean protein hydrolyzed by bromelain

Enzymatic bromelain mungbean meal protein hydrolysate (eb-MPH) was produced from mungbean meal protein isolate (MPI). Enzymatic bromelain, with a known protease activity of 98,652 (unit/g), was used at concentrations of 0, 2, 6, 10, 14 and 18% (w/w) and with hydrolysis times of 0.5, 3, 6, 12, and 24 h. The pH and temperature were controlled at 6.0 and 50 °C, respectively. It was found that the best treatment combination for eb-MPH production by response surface methodology (RSM) was 18% bromelain and a hydrolysis time of 3 h, resulting in the greatest degree of hydrolysis (% DH) and percent yield, with values of 61.04 and 45.63%, respectively. Results also showed that the phenylalanine, tyrosine and leucine contents of the optimally produced eb-MPH were 20.88, 14.50 and 10.93%, respectively. Twelve volatile compounds were identified using gas chromatography mass spectrometry in eb-MPH; benzaldehyde, 2-pentylfuran and furfural were the predominant odorants.     

Novel method for the production of pure glycinin from soybeans

A novel method for the purification of glycinin from soy meal is presented. The method is based on the isoelectric precipitation of glycinin by using carbon dioxide as a volatile precipitant. Gaseous CO(2) was pressurized into the protein solution, thus lowering the pH and initiating glycinin precipitation. Pressurization and, consequently, acidification were done in a slow and controlled manner, with the end point of pH 6.4. The acidity of the protein solution was well controlled via the pressure of gaseous CO(2). In this way simultaneous precipitation of other soybean proteins was prevented and very pure glycinin was obtained. Approximately 40% of the glycinin present in the protein solution was recovered with purity as high as 98%. The purification process was successfully performed on both small and large scales, without affecting glycinin purity.