大米蛋白酶水解物的抗氧化活性研究

利用Alcalase蛋白酶水解大米蛋白0.5-6 h.用pH-stat法测定其水解度,并在卵磷脂脂质氧化体系中.测定硫代巴比妥酸反应物质(TBARS),得出5 h的碱性蛋白水解产物具有较高的抗氧化活性.测定不同浓度下的大米蛋白水解产物(5 h)的还原能力和清除自由基能力,以及对大米蛋白水解物进行氨基酸分析.结果表明,随着酶解物浓度的增加,还原能力和清除DPPH能力也逐渐增大;而且证实这与蛋白水解后的结构以及水解产物中的短肽、氨基酸有关.     

球磨处理对鸡蛋清蛋白结构、性质及起泡性的影响

蛋清因具有优良的起泡性而在焙烤等食品加工领域中被广泛应用,因此,提升蛋清蛋白的起泡性有重要意义。本实验对鸡蛋清蛋白进行球磨处理,通过测定球磨处理后蛋清蛋白的游离巯基含量、表面疏水性、水解度、Zeta-电位和粒径,并分析其热力学性质、二级结构、起泡特性和微观结构,以研究球磨处理对蛋清蛋白结构性质及其起泡性的影响。结果表明：球磨处理后的蛋清蛋白粉随处理时间的延长起泡性无明显变化,泡沫稳定性呈现先升高后降低的趋势,且在球磨处理40 min时达到最大值（33.5 s）,比对照组提高近3 倍;此外,球磨处理可显著降低蛋清蛋白表面疏水性和热变性焓（ΔH）（P＜0.05）,显著提高水解度和降低Zeta-电位绝对值（P＜0.05）,并增大粒径,但对游离巯基含量和变性温度无显著影响（P＞0.05）;随着球磨时间的延长,蛋清蛋白β-折叠结构相对含量显著升高（P＜0.05）,α-螺旋结构相对含量先降低后升高,而β-转角结构相对含量先升高后降低;扫描电子显微镜结果显示球磨处理使蛋清蛋白呈现出零散碎片化和絮状。综上,适度球磨处理可以改变蛋清蛋白分子结构,进而提升其泡沫稳定性。     

大米蛋白的酶水解机制研究——Ⅱ酶水解过程中蛋白质的组分变化

凝胶色谱和高效液相色谱分析表明,大米蛋白在碱性蛋白酶Alcalase水解过程中,其可溶性蛋白组分的相对分子质量(Mr.)分布范围相对稳定,但Mr.1350u以下小分子组分的相对含量不断增加;十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)分析显示,随着酶水解反应的进行,Mr.为57、39、26、22ku的亚基逐渐消失,而29ku和13ku两个亚基的含量相应增加,且这两个亚基表现出可抵抗酶水解的特性。氨基酸分析表明,酶解后残余物蛋白中胱氨酸、蛋氨酸等含硫氨基酸的含量显著高于可溶性蛋白中的含量。圆二色光谱(CD)分析显示,酶水解前后大米蛋白的二级结构发生显著变化,在残余物蛋白质组分中自由回转的比例高达71.3%,α-螺旋结构完全消失。     

胶束化和碱溶酸沉提取羽扇豆蛋白的物化特性

利用胶束化和碱溶酸沉2 种提取方法从羽扇豆中提取羽扇豆蛋白,并对2 种方法提取羽扇豆蛋白的物化特性进行研究。结果表明：羽扇豆蛋白分子质量主要集中在18～90 kDa之间,是由多种分子质量的蛋白组成。碱溶酸沉提取羽扇豆蛋白的氨基酸总含量（66.2%）显著（P＜0.05）高于胶束化提取羽扇豆蛋白的氨基酸总含量（27.8%）。2 种提取方法提取羽扇豆蛋白的等电点pI接近5.0。胶束化提取羽扇豆蛋白的持水性显著（P＜0.05）高于碱溶液酸沉法提取的羽扇豆蛋白,而持油性无显著差异（P＞0.05）。碱溶酸沉法提取羽扇豆蛋白的起泡性在pH 2.0～6.0之间显著减小,在pH 6.0时起泡性最低,其泡沫稳定性的变化趋势与其起泡性类似。而胶束化提取羽扇豆蛋白的起泡性受pH值的影响不显著（P＞0.05）。胶束化提取的羽扇豆蛋白在pH 4.0时乳化性最小,然后随pH值的升高蛋白乳化活性也随之增加,2 种提取方法所提羽扇豆蛋白在pH 10.0时乳化活性最高。碱溶酸沉法提取的羽扇豆蛋白在pH 4.0时溶解度最小,而随着pH 4.0～10.0的增加,蛋白溶解度迅速增加。而胶束化提取羽扇豆蛋白的溶解度随pH 2.0～10.0的升高也逐渐增大。该研究为羽扇豆蛋白在食品工业中的开发与应用提供一定理论支持。     

大米浓缩蛋白限制性水解及其性质研究

以水解度为依据,比较了4种商品蛋白酶水解大米浓缩蛋白的效率.碱性蛋白酶具有最高的水解效率,其最佳水解条件为温度50℃～60℃,pH8.5,加酶量1%(E/S),底物浓度6%.考察了频率40 kHz,功率40 W～100 W的超声波对碱性蛋白酶水解的辅助作用.超声波在蛋白水解初期有一定加速度作用,不同的超声功率对水解速率影响并不显著.在蛋白水解水解度2%～10%范围内,用碱性蛋白酶制备了不同水解度的大米蛋白产品,并比较了他们溶解性、发泡性和持泡性以及口感.蛋白质的水解度与溶解性呈正相关,在水解度≥8%之后蛋白水解产物的开始出现苦味.蛋白质的持泡性随水解度的增加有一定改善,起泡性未见改善.     

骨蛋白水解物功能性质的研究

将骨蛋白利用碱性蛋白酶进行水解来达到改善骨蛋白的功能性质,以便于扩大骨蛋白在食品工业中的应用。本试验将制备的骨蛋白用碱性蛋白酶水解不同时间,用pH-Stat法测定其水解度,并测定不同时间的水解产物的持水性和持油性,并测定在不同的pH条件下的乳化性及乳化稳定性、起泡性以及起泡稳定性。试验结果表明,骨蛋白的水解物具有较好的持油性、乳化性及起泡性,尤其是3h的水解产物,但是乳化稳定性和起泡稳定性差。     

限制性酶解对大米蛋白结构、功能特性及体外抗氧化活性的影响

利用胰蛋白酶对大米蛋白进行限制性酶解,得到2%,4%和8%水解度的大米蛋白水解产物,探究不同水解程度对大米蛋白的结构、理化性质及体外活性的影响。结果表明,限制性酶解对大米蛋白的溶解性及乳化性有很大提升,当水解度低于4%时,酶解产物具有良好的热稳定性,然而,随着水解度的增大,其热稳定性呈下降趋势。因酶解作用,大米蛋白中β-转角含量升高,故具有更加舒展的二级结构。其表面呈疏松多孔的微观结构。相比于天然大米蛋白,酶解产物体外抗氧化活性显著提高,而且在高浓度时DPPH和ABTS自由基清除率与同浓度的BHT相当。限制性酶解作为一种温和、安全且高效的改性方法,可有效改善大米蛋白的功能特性,提高其体外抗氧化活性,以丰富大米蛋白资源的应用。     

酸法脱酰胺处理对大米蛋白功能特性的影响

采用HCl对大米蛋白进行脱酰胺处理,比较了脱酰胺处理前后大米蛋白溶解性、乳化性、起泡性及表面疏水性的变化。结果表明:脱酰胺处理过程中大米蛋白脱酰胺度和水解度均随反应时间增加而逐步增加,其功能性质均得到显著提高;中性条件下,脱酰胺处理4 h的大米蛋白溶解性提高了约7.86倍,乳化活性和乳化稳定性分别提高了4.67倍和3.39倍;起泡能力和泡沫稳定性在脱酰胺2 h后达到较佳,分别提高了5.92倍和1.92倍。此外,脱酰胺大米蛋白的表面疏水性发生明显下降。     

乌饭树树叶蓝黑色素对大米蛋白理化性质的影响

目的:研究乌饭树树叶蓝黑色素对大米蛋白组分结构性质和乳化性质的改善作用。方法:以乌饭树树叶为原料制备蓝黑色素,以乌米饭中大米蛋白为研究对象,模拟乌米饭染色过程并分析染色前后大米蛋白结构性质和乳化性质的变化。结果:乌饭树树叶蓝黑色素可与大米蛋白形成二元复合物,导致色度L^*值和b^*值降低,复合物颜色向蓝黑色转变。色素—蛋白复合物溶解度增加,而持油性显著降低。利用圆二色光谱分析大米蛋白二级结构,发现其α-螺旋与β-折叠的相对含量下降,β-转角和无规卷曲的相对含量上升。当色素质量分数低于1.5%时,起泡稳定性升高,而起泡性、乳化性和乳化稳定性降低。当色素质量分数高于1.5%后,起泡稳定性下降,起泡性、乳化性和乳化稳定性升高。结论:乌饭树树叶蓝黑色素通过改变大米蛋白二级结构可改善其乳化性质。     

燕麦蛋白耦合异源共架技术对大米蛋白水溶性的影响

为了通过燕麦蛋白耦合异源共架技术提升大米蛋白在水中的溶解特性,本研究以不同比例的大米蛋白和燕麦蛋白为原料,将两种蛋白在pH12的环境下混合,再恢复至中性进行共架。测定蛋白的溶解性并通过分析十二烷基磺酸钠-聚丙烯酰胺凝胶电泳、粒径、微观形态、乳化特性、起泡特性、傅里叶红外光谱等探究其作用机理。结果显示,大米-燕麦蛋白的溶解度显著高于大米蛋白（8.49%±1.53%）(P<0.05),两者比例为1:0.6时达到最大值93.07%±2.15%。经过异源共架处理的大米-燕麦蛋白复合物粒径为0.1μm左右,同时结构更加松散,大米蛋白和燕麦蛋白之间的疏水相互作用和氢键也发生了变化,使得大米蛋白的溶解度增加,进一步提升乳化特性和起泡特性。综上,燕麦蛋白耦合异源共架技术是一种可以解决大米蛋白水溶性差的可靠手段。     

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.     

Antioxidant activity and functional properties of porcine plasma protein hydrolysate as influenced by the degree of hydrolysis

Antioxidant activity and functional properties of porcine blood plasma protein hydrolysates (PPH) prepared with Alcalase at 6.2%, 12.7% and 17.6% of degree of hydrolysis (DH) were investigated. The PPH showed stronger radical-scavenging ability and possessed stronger Cu²⁺-chelation ability and a reducing power compared to non-hydrolysed plasma protein (P < 0.05). The antioxidant activity of PPH, indicated by thiobarbituric acid-reactive substance (TBARS) values in a liposome-oxidising system, increased with increasing DH (P < 0.05). The Alcalase hydrolysis increased protein solubility from its original 68.46–81.79% (non-hydrolysed) to 82.95–94.94% (hydrolysed) over a broad pH range (3.0–8.0). However, hydrolysis decreased surface hydrophobicity and suppressed emulsifying and foaming capacity of the plasma protein. To identify antioxidant peptide, PPH was subjected to ultrafiltration, ion-exchange chromatography and reverse-phase high performance liquid chromatography (RP-HPLC), and the amino acid sequences of isolated peptides were determined by liquid chromatography/tendem mass spectrometry (LC–MS/MS). The peptide with the strongest antioxidant activity had the amino acid sequence of His-Asn-Gly-Asn. The results indicated that PPH could be used as a novel antioxidant but may be of limited utility as an emulsifying or foaming agent.     

Antioxidant Activities of Rapeseed Protein Hydrolysates

Rapeseed protein hydrolysates (RPH) were obtained by enzymatic hydrolysis of rapeseed protein using Alcalase 2.4 L FG. The degree of hydrolysis (DH) of RPH was about 25% using pH-stat method. The antioxidant activities of RPH were investigated by employing several in vitro assay, including the 1,1-diphenyl-2-picrylhydrazyl (DPPH)/superoxide/hydroxyl radical scavenging assays, and reducing power assay. RPH showed scavenging activity against free radicals such as DPPH, superoxide, and hydroxyl radicals. The radical scavenging effect was in a dose-dependent manner, and the EC₅₀ values for DPPH, superoxide, and hydroxyl radicals were found to be 0.71, 1.05, and 4.92 mg/mL, respectively. In addition, the RPH also exhibited notable reducing power, which was 0.51 at 2.00 mg/mL. The data obtained by in vitro systems obviously established the antioxidant potency of RPH. Combined with the results of the amino acid profiles, RPH were believed to have high nutritive value in addition to antioxidant activities.     

Improvement of functional properties and antioxidant activities of cuttlefish (Sepia officinalis) muscle proteins hydrolyzed by Bacillus mojavensis A21 proteases

Functional properties and antioxidant activities of cuttlefish (Sepia officinalis) muscle protein hydrolysates, with different degrees of hydrolysis (DH from 7.3% to 18.8%), obtained by treatment with Bacillus mojavensis A21 alkaline proteases were investigated. Protein contents for all freeze-dried cuttlefish muscle protein hydrolysates (CMPHs) ranged from 80% to 86%. For the functional properties, hydrolysis by A21 proteases increased (p < 0.05) protein solubility to above 78% over a wide pH range (2.0–11.0). However, the interfacial activities (emulsion activity index, emulsion stability index, foaming capacity and foaming stability) decreased with the increase of the DH. All CMPHs exhibited significant metal chelating activity and DPPH free radical-scavenging activity, and inhibited linoleic acid peroxidation. Antioxidant properties of protein hydrolysates increased with protein hydrolysis and the highest activities were obtained at DH of 16%. The IC₅₀ values for DPPH radical-scavenging and metal chelating activities were found to be 0.52 ± 0.01 mg/ml and 0.67 ± 0.13 mg/ml. The obtained results suggested that functional properties and antioxidant activities of cuttlefish muscle protein hydrolysates were influenced by the degree of hydrolysis.The composition of amino acids of undigested and hydrolyzed proteins was determined. CMPHs have a high percentage of essential amino acids such as arginine, lysine, histidine and leucine. They have a high nutritional value and could be used as supplement to poorly balanced dietary proteins.     

碱性蛋白酶限制性酶解对蓝圆鲹分离蛋白功能特性的影响

以蓝圆鲹(Decapterus maruadsi)分离蛋白为原料,采用碱性蛋白酶对其进行限制性酶解,研究水解度(degree of hydrolysis,DH)对分离蛋白酶解产物溶解性、持油力、乳化性与起泡性等功能特性的影响.结果表明,碱性蛋白酶酶解产物的相对分子质量显著下降.酶解可有效提高分离蛋白的溶解性,其溶解度随...     

鲤鱼鱼肉蛋白酶水解物抗氧化性及功能特性

采用碱性蛋白酶对鲤鱼鱼肉蛋白进行酶解,制备不同水解度的水解物。测定水解物的抗氧化活性以及不同pH值条件下水解物的功能特性。结果表明:随着水解度的逐渐升高,水解物的抑制脂质氧化能力、D PP H自由基清除能力、还原能力以及金属离子(Cu2+和Fe2+)螯合能力逐渐增加(P<0.05);同时,水解物的溶解性、乳化性和起泡性都在pH值为4.0(等电点)时达到最低,而后溶解性和乳化性随着pH值升高而增大(P<0.05),而起泡性随着pH值的升高先上升后又下降。因此,鲤鱼鱼肉蛋白碱性蛋白酶水解物可以提高蛋白质的抗氧化活性和溶解性,但是较高的水解度会在一定程度上降低其乳化性和起泡性。     

乙醇对谷朊粉酶解产物的苦味影响研究

为了探寻降低谷朊粉酶解产物苦味的方法,通过添加不同浓度的乙醇(5%、10%和15%),研究其对碱性蛋白酶酶解产物苦味值、游离氨基酸和相对分子质量分布的影响,以及在乙醇存在下碱性蛋白酶、中性蛋白酶以及风味蛋白酶3种复配酶解体系所制得的酶解产物的苦味值和相对分子质量分布变化。结果表明:加入乙醇后,在相同的水解度下(DH),碱性蛋白酶的酶解产物苦味值降低,并与加入乙醇的浓度呈负相关。添加乙醇后,相对分子质量小于1 000的组分含量显著(P0.05)降低,游离疏水性氨基酸(Pro,Ile,Phe和Met)显著(P0.05)增加。添加乙醇后,3种复配酶解体系酶解产物苦味降低,相对分子质量小于1 000的组分含量显著(P0.05)降低。与添加碱性蛋白酶单一酶解相比,在相同的水解度下3种酶复配酶解产物的苦味值进一步降低。添加低浓度乙醇对3种酶的活性影响较小。     

大米蛋白酶解物—大豆苷元复合纳米粒子的构建

以大米蛋白为原料,经胰蛋白酶酶解后制备纳米载体,以大豆苷元为输送对象,构建了大米蛋白酶解物—大豆苷元的纳米输送体系,通过试验考察了大米蛋白水解度、大米蛋白酶解物质量浓度与大豆苷元质量浓度对制备的复合纳米粒子性质的影响。在大米蛋白水解度为12%,酶解物质量浓度为7.5 mg/mL,大豆苷元质量浓度为0.75 mg/mL的条件下制备的大米蛋白酶解物—大豆苷元复合纳米粒子,其平均粒径449.71 nm、多分散系数0.23,Zeta-电位-22.30 mV,稳定性较好;大豆苷元的包封率为60.07%,荷载量为6.01 mg/g,证明大米蛋白酶解物具有构建纳米载体荷载大豆苷元的可行性。     

Controlled enzymatic hydrolysis on characteristic and antioxidant properties of soybean protein isolate-maltodextrin conjugates

This study aimed to investigate the effect of limited hydrolysis on conformational and antioxidant properties of soy protein isolate-maltodextrin (SPI-Md) conjugates. Extrinsic fluorescence analysis showed unfolding of the protein molecule and exposure of hydrophobic groups in SPI-Md conjugate hydrolysates. Free amino acid analysis showed that, the contents of hydrophobic amino acids in SPI-Md conjugates increased after hydrolysis. The contents of leucine, isoleucine, phenylalanine increased from 0.32, 0.30 and 0.54 to 1.36, 1.86 and 2.60, respectively, when the hydrolysis degree (DH) gradually increased from 0% to 5.7%. The FT-IR spectrum showed that C = O absorption of the amide group formed by glycosylation continued unabated after limited hydrolysis (DH 2.9%). The glycated SPI products showed good reducing power and superior resistance to lipid oxidation (34%, 12 mg mL^?1), whereas the limit hydrolysates (DH 2.9%) of SPI-Md conjugates showed more ef?cient radical-scavenging capacity (89.5%, at 12 mg mL^?1) and iron-chelation activity (91.3%, at 12 mg mL^?1). Results of this study indicated that, slight enzymatic hydrolysis (DH 0–2.9%) could help partially unfolding the globular structure of SPI-Md conjugates without deteriorating amide bonds and had a positive effect on their antioxidant properties.     

菜籽分离蛋白分子质量分布及酶解条件的研究

以脱脂"双低"油菜籽为原料,利用碱溶解酸沉淀法提取菜籽分离蛋白;用SDS-PAGE凝胶电泳研究菜籽蛋白的分子质量的组成;以水解度和氮回收率为考察指标,用响应面分析法拟合了Alcalase 2.4L酶解菜籽蛋白成菜籽肽的二次多项数学模型,优化了酶解菜籽分离蛋白的工艺参数。SDS-PAGE凝胶电泳研究表明利用碱溶解酸沉淀提取的菜籽分离蛋白主要是2S清蛋白。响应面分析法研究的试验结果表明,酶的使用量、pH、酶解温度、酶的使用量与pH的交互作用对Alcalase 2.4L酶解菜籽蛋白的水解度和氮回收率的影响均显著(P0.05)。通过求解菜籽肽的二次多项数学模型的逆矩阵方程,可得Alcalase 2.4L水解菜籽分离蛋白的最佳条件为:酶的使用量0.05 Au/g,pH 9.0,酶解温度54℃;在此酶解条件下,菜籽分离蛋白浓度为5%时,水解5 h,所得的水解度和氮回收率分别为35.12%及52.96%。