天然低共熔溶剂提取米糠酯酶工艺优化及其纯化与酶学性质研究

目的：采用天然低共熔溶剂（Natural deep eutectic solvents,NADES）提取米糠酯酶并优化提取工艺参数,纯化米糠酯酶获得高纯度米糠酯酶,并探究其酶学特性。方法：合成了10种不同的NADES、选用了水浴搅拌和超声两种处理方式进行筛选,然后对提取效果最佳的NADES和方法进行单因素和响应面优化,参数包括料液比、含水量、提取温度和提取时间;将最优提取工艺得到的粗酶液通过阴离子交换树脂DEAE纯化,然后进行酶学性质的探究。结果：最佳提取米糠酯酶的工艺为在含有5%水分的脯氨酸-甘油（摩尔比1:2）溶剂体系中,水浴搅拌3.0 h,温度74.0℃,米糠和溶剂料液比为9:30,此时米糠酯酶活力为2.96 U。通过阴离子交换树脂DEAE纯化,获得了纯度较高的米糠酯酶,纯化倍数为1.74倍,回收率为69.40%,分子量约为35 kDa,对硝基苯酚乙酯为最适底物,最适温度和pH分别是40.0℃、pH8.0,在30.0～40.0℃和pH7.0～9.0时米糠酯酶均保持良好的稳定性。氯化胆碱-甘油、脯氨酸-甘油两种NADES对米糠酯酶的活力有较好的维持作用。结论：天然低共熔溶剂提取方式...     

超声波辅助酸性天然低共熔溶剂提取黑果腺肋花楸花青素及其稳定性和抗氧化活性分析

建立一种绿色、高效的超声波辅助酸性天然低共熔溶剂提取黑果腺肋花楸花青素的新方法,利用人工神经网络和遗传算法优化提取条件,并研究花青素提取物的稳定性和抗氧化活性。以甜菜碱和有机酸为氢键受体和氢键供体,制备了一系列酸性天然低共熔溶剂,并对其密度、粘度、pH理化性质进行了测定,通过红外光谱研究了天然低共熔溶剂的结构和形成机理,利用人工神经网络结合遗传算法优化了最佳提取条件,并评价了花青素提取物的光稳定性、热稳定性和抗氧化活性。结果表明,甜菜碱和乳酸通过氢键相互作用形成的天然低共熔溶剂具有密度低(1.19)、粘度小(24.75 mPa·s)、pH低(2.89)的特点,其最佳提取条件为：以甜菜碱和乳酸制备天然低共熔溶剂,摩尔比1:3,含水量为32%,超声功率124 W,超声时间24 min,初始超声温度32℃。在此最佳条件下,花青素的提取率达到23.62 mg/g。与传统溶剂和其它方法相比,本方法绿色高效,操作简单。稳定性和抗氧化实验结果显示,光照会加速提取物中花青素的降解,当温度大于50℃时,花青素热降解加速,一级动力学降解常数k>0.0234。当质量浓度为200μg/mL时,花青素提取...     

天然低共熔溶剂萃取酚类物质研究进展

文章综述了天然低共熔溶剂(NADES)对酚类物质提取分离研究现状,总结了影响提取效率的主要因素,并对未来发展方向作出了展望。     

紫马铃薯花色苷提取工艺优化及稳定性、抗氧化活性分析

以甜菜碱为氢键受体,有机酸、糖基和醇基分别为氢键供体制备天然绿色的低共熔溶剂,基于微波辅助提取法,通过单因素实验并结合响应面分析对紫马铃薯花色苷提取工艺进行优化。主要考察了微波时间、微波功率、溶剂含水量、溶剂摩尔比对紫马铃薯花色苷含量的影响。同时比较低共熔溶剂与常规溶剂提取对紫马铃薯花色苷在不同温度、光照条件下的稳定性,以及DPPH、ABTS+、OH自由基清除率评价体外抗氧化能力。结果表明,以甜菜碱和柠檬酸制备酸性低共熔溶剂,摩尔比1:2.1,含水量为28.6%,在微波功率800 W,微波时间28 s条件下,紫马铃薯花色苷含量可达到228.658±1.241 mg/100 g,较常规提取工艺含量提高了56.92%。此外通过低共熔溶剂提取所得花色苷在不同光照、温度条件下稳定性均显著提高。其中太阳光对紫马铃薯花色苷影响最大,避光情况下低共熔溶剂提取花色苷保存率可达90%以上,常规溶剂提取花色苷保存率为82.78%,此外,花色苷含量也随着温度的增加不断降低,二者保存率均明显下降。抗氧化能力结果表明,低共熔溶剂提取所得花色苷抗氧化能力更强,其清除自由基能力IC₅₀值均小于...     

天然低共熔溶剂提取柑橘籽类柠檬苦素的工艺优化

为建立柑橘籽中柠檬苦素类化合物的绿色环保提取方法,该研究制备了13种天然低共熔溶剂,并对其理化性质进行了测定。采用超声辅助提取法从柑橘籽中提取柠檬苦素类化合物,并应用超高效液相色谱检测3种主要的柠檬苦素类化合物(柠檬苦素、诺米林和黄柏酮)。与传统乙醇溶剂(体积分数80%)相比,由氯化胆碱和甲酸(物质的量之比1∶4)组成的天然低共熔溶剂具有较好的提取效率。在此基础上,采用响应面法对含水量、液固比、提取温度和提取时间进行优化。结果显示,含水量12%(体积分数)、液固比10∶1(mL∶g)、提取温度60℃、提取时间34 min为最佳提取工艺,类柠檬苦素得率为13.87 mg/g。该研究表明,制备的天然低共熔溶剂可以作为从柑橘籽中提取类柠檬苦素的替代溶剂,为进一步应用天然低共熔溶剂从天然产物中提取生物活性物质提供参考。     

天然低共熔溶剂提取油茶蒲中植物多酚的研究

以油茶蒲为原料,对照传统的水和60%乙醇溶液,选择几种天然低共熔溶剂提取油茶蒲中的植物多酚,考察了天然低共熔溶剂对多酚得率的影响。通过单因素试验和响应面试验对天然低共熔溶剂提取油茶蒲中多酚的工艺条件进行了研究和优化。结果发现:天然低共熔溶剂具有比传统溶剂更好的提取效果;油茶蒲中多酚提取的最优条件为提取溶剂柠檬酸-氯化胆碱(摩尔比1∶3)、提取时间37 min、提取温度81℃、液料比42∶1,在最优条件下多酚得率为0. 932 1%。     

低共熔溶剂提取的黄精多糖性质分析

为研究低共熔溶剂提取条件对黄精多糖性质及体外抗氧化活性的影响,以鸡头黄精为原料,对不同条件下低共熔溶剂提取得到的黄精多糖进行相对分子量、单糖组成等基本性质和体外抗氧化活性的测定。结果显示,相比于传统水提醇沉法,采用低共熔溶剂法提取黄精多糖,70 ℃时得率为18%,提高了36%,所得多糖的相对分子量变小且半乳糖含量升高;100 ℃提取的多糖相对分子量更小,且主要成分为葡萄糖。羟基自由基与DPPH自由基清除率、总抗氧化能力测定结果均表明,采用低共熔熔剂在70 ℃条件下提取的黄精多糖体外抗氧化能力明显高于传统水提醇沉法和低共熔溶剂法在100 ℃条件下提取的黄精多糖。当浓度为3.0 mg/mL时,采用低共熔熔剂在70 ℃提取的黄精多糖总抗氧化能力为4.5 U/mL,是水提多糖的4.3倍,是100 ℃条件提取黄精多糖的7.4倍。低共熔溶剂对黄精多糖具有降低分子量、提高抗氧化性等效果,研究结果可以为低共熔溶剂在多糖提取方面的应用提供参考。     

低共熔溶剂提取桂花黄酮的工艺优化

为了探索一种高效、环保的桂花黄酮提取方法,本文设计并制备了6种低共熔溶剂,通过比较醇提、冻融、超声波及微波四种技术,确定了超声波辅助-低共熔溶剂的提取工艺。通过单因素试验考察液料比、摩尔比、含水量、超声功率以及超声时间对桂花黄酮提取量的影响。在单因素的基础上,采用响应面法对提取工艺进一步优化。结果表明：桂花黄酮最优提取工艺为超声波辅助-三元低共熔溶剂提取（氯化胆碱/山梨醇/1,2-丙二醇）;最佳工艺条件为：液料比60:1 (mL/g),氯化胆碱:山梨醇:1,2-丙二醇=2:1:4,含水量62 mol,超声波功率150 W,超声时间16 min,在此条件下提取量达到了10.06 mg/g,比传统醇提法提高了8.93 mg/g,证实超声波辅助-绿色低共熔溶剂技术提取桂花黄酮的高效性。实验结果与响应面模型预测值接近,证实模型的有效性。本研究为低共熔溶剂在天然产物绿色提取方面的应用提供参考。     

微波辅助低共熔溶剂提取桑葚果渣花青素的工艺研究

以新型绿色低共熔溶剂为提取剂,采用微波辅助法从桑葚果渣中提取花青素。在单因素试验结果基础上,应用响应面分析法对影响花青素提取量的提取条件进行优化。结果表明,桑葚果渣花青素最佳提取工艺条件为:以含水量为40%的氯化胆碱/1,2-丙二醇低共熔溶剂为最佳提取剂,料液比为1∶50,微波功率为600W,微波温度为40℃,微波时间为40s。在此条件下,花青素提取量为35.97mg/g。低共熔溶剂是一种绿色高效的提取剂,可用于生物活性成分的高效提取。     

低共熔溶剂提取桂花黄酮的工艺优化

为了探索一种高效、环保的桂花黄酮提取方法,本文设计并制备了6种低共熔溶剂,通过比较醇提、冻融、超声波及微波四种技术,确定了超声波辅助-低共熔溶剂的提取工艺。通过单因素试验考察液料比、摩尔比、含水量、超声功率以及超声时间对桂花黄酮提取量的影响。在单因素的基础上,采用响应面法对提取工艺进一步优化。结果表明:桂花黄酮最优提取工艺为超声波辅助-三元低共熔溶剂提取(氯化胆碱/山梨醇/1,2-丙二醇);最佳工艺条件为:液料比60:1 (mL/g),氯化胆碱:山梨醇:1,2-丙二醇=2:1:4,含水量62 mol,超声波功率150 W,超声时间16 min,在此条件下提取量达到了10.06 mg/g,比传统醇提法提高了8.93 mg/g,证实超声波辅助-绿色低共熔溶剂技术提取桂花黄酮的高效性。实验结果与响应面模型预测值接近,证实模型的有效性。本研究为低共熔溶剂在天然产物绿色提取方面的应用提供参考。     

大豆蛋白溶解性研究

该文概述大豆蛋白溶解特性及其与一般物质溶解差异,介绍提高大豆蛋白溶解性改性方法及研究现状,对比不同改性增溶方法优、缺点,并提出今后大豆蛋白改性研究方向。     

Short communication: Muscle protein synthetic response to microparticulated whey protein in middle-aged men

Whey protein concentrate (WPC) is a high-quality dairy ingredient that is often included in formulated food products designed to stimulate muscle anabolism. Whey protein concentrate can be affected by UHT processing, and its sensory properties are not compatible with some formulated food products. Microparticulated WPC (mWPC) is a novel ingredient that is resistant to heat treatment and has enhanced sensory properties. When 16 healthy middle-aged men consumed 20 g of either WPC or mWPC, both proteins increased plasma essential AA and leucine concentrations with no detectable difference in curve kinetics. Myofibrillar protein synthesis was increased in both groups for 90 min after ingestion with no difference between groups. Ingestion of mWPC resulted in a muscle anabolic response that was equivalent to that of WPC over the full 210-min measurement period. Formulated products incorporating mWPC or standard WPC would provoke equivalent anabolic responses.     

Co-precipitation of whey and pea protein – indication of interactions

The aim was to optimise the yield of co-precipitation of whey protein isolate (WPI) and pea protein isolate (PPI) and compare co-precipitates and protein blends with respect to solubility. The yield of co-precipitates was tested with different protein ratios of WPI and PPI in combination with different temperatures and acid precipitation (pH 4.6). The highest precipitation yield was obtained at protein ratios WPI < PPI, high temperature and alkaline protein solvation. The solubility was measured by an instability index and absorption spectroscopy of re-suspended precipitated proteins at pH 3, 7 and 11.5. Co-precipitates had significantly lower solubility than protein blends. Protein ratios WPI > PPI, low precipitation temperature and high pH showed the highest solubility. Differences in protein composition between co-precipitates and protein blends were observed with SDS-PAGE and matrix-assisted laser desorption ionisation time-of-flight, and indicated different protein–protein interaction in samples, which needs further investigations.     

细菌中蛋白质样品制备方法研究进展

细菌在生长繁殖时,细菌蛋白的表达受到环境影响而存在较大差异,使得细菌蛋白表达具有复杂性。在食品生产加工过程中可能会受到致病菌污染,细菌产生的内毒素和外毒素均会对人体健康构成威胁,因此需要高灵敏度和高特异性的检测方法来定量分析和鉴定食品中的细菌毒素。蛋白组学方法可以揭示细菌蛋白组成及其潜在的生物学功能,感染过程中菌体蛋白表达变化和致病机制理。其中,细菌蛋白样品的前处理作为细菌蛋白组学研究的关键步骤,直接影响鉴定的细菌蛋白质量。本文主要阐述目前细菌蛋白组学研究中菌体蛋白和胞外蛋白样品制备方法及其对蛋白鉴定和定量的影响,将为更深入的开展细菌蛋白组学研究提供参考。     

Physicochemical and Functional Properties of Soy Protein Substrates Modified by Low Levels of Protease Hydrolysis

ABSTRACT: Endo-protease treatments achieving low degrees of hydrolysis (DH 2% and 4%) were used to improve functional properties of hexane-extracted soy flour (HESF), extruded-expelled partially defatted soy flour (EESF), ethanol-washed soy protein concentrate (SPC), and soy protein isolate (SPI). These substrates had protein dispersibility indices ranging from 11% to 89%. Functional properties, including solubility profile (pH 3 to 7), emul-sification capacity and stability, foaming capacity and stability, and apparent viscosity were determined and related to surface hydrophobicity and peptide profiles of the hydrolysates. Protein solubilities of all substrates increased as DH increased. Emulsification capacity and hydrophobicity values of the enzyme-modified HESF and EESF decreased after hydrolysis, whereas these values increased for SPC and SPI. Emulsion stability was improved for all 4% DH hydrolysates. Hydrolyzed SPC had lower foaming capacity and stability. For substrates other than SPC, foaming properties were different depending on DH. Hydrolysis significantly decreased the apparent viscosities regardless of substrate. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) indicated differences in the molecular weight profiles of the hydrolysates. HESF and EESF, which had high proportions of native-state proteins, showed minor changes in the peptide profile due to hydrolysis compared with SPC and SPI.     

几种新型花生蛋白产品的生产

本文从花生蛋白质利用的角度介绍了几种新型花生蛋白产品的加工工艺，针对我国人民膳食结构中蛋白质的摄入水平较低的实际情况，论述了花生蛋白开发利用的必要性，展望了花生蛋白开发利用的广阔前景。     

大豆分离蛋白改性对其功能性质影响

大豆分离蛋白是大豆蛋白最为精制形式,广泛应用于食品工业,并在不同产品中表现出不同功能。该文综述近年来大豆分离蛋白物理、化学、酶法及基因工程改性对其功能性质影响,经不同方式改性可产生合适功能性质,从而拓宽大豆分离蛋白在食品工业中应用。     

Formulation,process conditions,and biological evaluation of dairy mixed gels containing fava bean and milk proteins: Effect on protein retention in growing young rats

Food formulation and process conditions can indirectly influence AA digestibility and bioavailability. Here we investigated the effects of formulation and process conditions used in the manufacture of novel blended dairy gels (called “mixed gels” here) containing fava bean (Vicia faba) globular proteins on both protein composition and metabolism when given to young rats. Three mixed dairy gels containing casein micelles and fava bean proteins were produced either by chemical acidification (A) with glucono-δ-lactone (GDL) or by lactic acid fermentation. Fermented gels containing casein and fava bean proteins were produced without (F) or with (FW) whey proteins. The AA composition of mixed gels was evaluated. The electrophoretic patterns of mixed protein gels analyzed by densitometry evidenced heat denaturation and aggregation via disulfide bonds of fava bean 11S legumin that could aggregate upon heating of the mixtures before gelation. Moreover, fermented gels showed no particular protein proteolysis compared with gel obtained by GDL-induced acidification. Kinetics of acidification were also evaluated. The pH decreased rapidly during gelation of GDL-induced acid gel compared with fermented gel. Freeze-dried F, A, and FW mixed gels were then fed to 30 young (1 mo old) male Wistar rats for 21 d (n = 10/diet). Fermented mixed gels significantly increased protein efficiency ratio (+58%) and lean mass (+26%), particularly muscle mass (+9%), and muscle protein content (+15%) compared with GDL-induced acid gel. Furthermore, F and FW formulas led to significantly higher apparent digestibility and true digestibility (+7%) than A formula. Blending fava bean, casein, and whey proteins in the fermented gel FW resulted in 10% higher leucine content and significantly higher protein retention in young rats (+7% and +28%) than the F and A mixed gels, respectively. Based on protein gain in young rats, the fermented fava bean, casein, and whey mixed proteins gel was the most promising candidate for further development of mixed protein gels with enhanced nutritional benefits.