桃仁多肽的酶法制备及其抗氧化活性分析

以桃仁蛋白为原料,对响应面法优化多肽制备条件和酶解多肽各组分的抗氧化活性进行了研究。结果表明:以底物浓度3.5%、pH 10.4、加酶量4 200 U/g、酶解时间4.9 h条件下进行碱性蛋白酶酶解,可获得桃仁蛋白的最高水解度(61.12±0.7)%和多肽得率(67.91±0.5)%。桃仁蛋白及其酶解物的SDS-PAGE凝胶电泳图表明,酶解后的多肽分子质量大部分小于12 000 u。酶解液(PPH)及其超滤分离所得4组分分别采用DPPH自由基清除能力、ABTS自由基清除能力、亚硝酸根离子清除能力和总抗氧化能力4个体外抗氧化活性测试,结果显示不同分子质量组分间的抗氧化活性存在显著性差异(P0.05),分子质量越小,其抗氧活性越强。RP-HPLC分析表明,抗氧化活性最强的P-IV组主要由9种具有一定疏水性的多肽组成。     

冷冻及加热法去除ε-聚赖氨酸发酵液中的蛋白

ε-聚赖氨酸（ε-PL）发酵液中蛋白的去除是ε-PL提取工作中重要的一环。以ε-PL和蛋白的相对质量含量为衡量指标,比较了冷冻和加热两种去除蛋白的方法,结果表明加热处理去蛋白的效果要优于冷冻处理。优化了加热处理条件：pH6.0、温度80℃、加热时间15min。在此条件下,蛋白去除率达83%且ε-PL基本没有损失。采用SDS-PAGE电泳对热处理去蛋白效果进行了鉴定,结果表明发酵液中蛋白分子量分布在6.64万以下,只有少量1.43万以下的小分子蛋白不能加热去除。研究建立的加热处理ε-PL发酵液的方法对蛋白的去除具有一定的指导意义。     

应用SDS-PAGE法评价乳基婴幼儿配方食品中有效蛋白质组分

利用SDS-PAGE法评价乳基婴幼儿配方食品中的有效蛋白组分,优化了样品前处理方法,采用Bio-Rad Quantity One软件对电泳谱图进行数据分析,实现牛乳蛋白组分的定性和半定量测定。采用该法对我国市售婴幼儿配方食品中的牛乳蛋白有效成分进行了质量评价分析,发现0～12月龄婴儿配方食品基本符合GB 10765-2010中对乳清蛋白不得低于60%总蛋白的规定;0～6月龄Ⅰ段婴儿配方食品中的乳清蛋白含量最高,乳清蛋白/酪蛋白为1.57～2.12(61∶39～67∶33);6～12月龄的Ⅱ段较大婴儿配方食品中乳清蛋白相对Ⅰ段少,二者比例为1.06～1.55(52∶48～61∶39);而在12月龄以上的Ⅲ段幼儿配方食品中二者比例为0.65～0.71(40∶60～42∶58)。虽然Ⅰ段婴儿配方食品乳清蛋白/酪蛋白符合标准,但是在母乳中占主要成分的α-LA(α-乳白蛋白)在Ⅰ段婴儿配方食品中含量偏低(19.1%～24.1%),母乳中不存在的α-CN(α-酪蛋白)在Ⅰ段婴儿配方食品中却占11.9%～17.8%,另外还发现Ig G(免疫球蛋白G)在乳基婴幼儿配方食品中的含量偏低。     

不同菌龄酿酒酵母细胞壁蛋白差异性分析

以酿酒酵母为研究对象,比较了完整细胞提取法、稀碱缓冲液提取法及溶菌酶和β-葡聚糖酶复合酶法等三种酵母菌细胞壁蛋白提取方法,分析了不同菌龄酵母细胞壁差异性蛋白。结果表明:溶菌酶和β-葡聚糖酶复合酶液水解纯化好细胞壁提取蛋白的方法具有所得胞壁蛋白条带较多,且纯度较高的优点,确定了此方法为提取酵母细胞壁蛋白的最佳提取方法。同时,通过SDS-PAGE电泳分析发现,不同菌龄酵母细胞壁蛋白存在着较大的差异性,并确定了分子质量在36 ku、17 ku和12 ku为不同酵母代数细胞壁的3个主要差异性蛋白,其中36 ku、17 ku处条带蛋白随着菌龄的增加酵母细胞壁蛋白表达量逐渐减少,而12 ku处条带蛋白随着菌龄的增加酵母细胞壁蛋白表达量逐渐增加。     

The effects of high oxygen modified atmosphere packaging on protein oxidation of bovine M. longissimus dorsi muscle during chilled storage

Modified atmosphere packaging (MAP) extends the shelf life of beef, especially in the context of visual colour. High O₂ MAP (70-80%) may cause quality deterioration through lipid and protein oxidation, the latter linked to a reduction in meat tenderness. The objective of this study was to investigate the effects of MAP in comparison to vacuum packaging (VP) on protein oxidation and subsequent tenderness of LD (M. longissimus dorsi) beef steaks during chilled storage (4 °C) of periods up to 14 days. Steaks were analysed for carbonyl content, free thiol groups, drip loss, Warner-Bratzler shear force (WBSF) and SDS-PAGE of extracted myofibrillar proteins. Free thiol groups were lower in high O₂ MAP (80% O₂/20% CO₂) steaks, after 14 days of storage, indicating protein oxidation compared to VP steaks. SDS-PAGE (non reducing conditions) showed the presence of high molecular weight cross linked myosin heavy chain aggregates in the high O₂ atmosphere packaged steaks, which were absent in VP steaks.     

Studies on the biosorption of Pb(II) ions using Pseudomonas putida

ABSTRACT

The complete removal of Pb(II) was achieved by intact Pseudomonas putida cells. The biosorption isotherm exhibited Langmuirian behaviour and followed pseudo-second-order rate kinetics. The standard Gibbs free energy change (?G°) for the biosorption of Pb(II) ions was found to be ?26.4 kJ mol^?1, attesting to a chemisorption process. Thermolysis of P. putida cells improved the Pb(II) binding capacity by around 27%. All the four components tested, namely DNA, protein, polysaccharide and lipid, were found to contribute to the uptake of Pb(II) ions. The possible mechanisms of Pb(II) binding by P. putida have been delineated.     

Gamma irradiation of cowpea (Vigna unguiculata L. Walp) flours and pastes: Effects on functional,thermal and molecular properties of isolated proteins

In a previous study irradiation of cowpea flours and pastes at medium (10 kGy) and high (50 kGy) doses resulted in significant changes in protein-related functional properties. To understand some of the effects of gamma irradiation on cowpea proteins in particular, we isolated proteins from cowpea flours (FPC) and pastes (PPC) treated with gamma irradiation at 2, 10, and 50 kGy and analyzed their functional, thermal and molecular properties. Nitrogen solubility index of both FPC and PPC decreased, whereas oil absorption and emulsion capacities increased significantly with increasing irradiation dose. Differential scanning calorimetry showed decreases in transition temperatures (T_d) and enthalpies (ΔH), indicating a progressive denaturation of cowpea proteins with increasing irradiation. Size exclusion HPLC revealed increases in protein molecular weights, probably due to protein–protein cross-linking with irradiation in a dose-dependent manner. Reducing SDS-PAGE of FPC and PPC samples seems to suggest that the contribution of disulphide bonds to irradiation-induced cowpea protein–protein cross-linking is small.     

Inhibition of biofilm formation of Pseudomonas aeruginosa by an acylated homoserine lactones-containing culture extract

The objective of this study was to examine the effect of an acylated homoserine lactones-containing culture extract (AHL-CCE) on bacterial growth, enzyme activity and biofilm formation of Pseudomonas aeruginosa. Chicken breast muscle broth in which P. aeruginosa had been grown for 24 h at 20 °C was extracted with acidified ethyl acetate, the solvent was evaporated and the residue was dissolved in water and heated at 100 °C for 15 min. Thin-layer chromatography combined with Agrobacterium tumefaciens KYC55-based bioassays indicated the presence of C₄-, C₆-, C₈- and C₁₂-homoserine lactones in the extract. The addition of AHL-CCE to a culture of P. aeruginosa caused the bacteria to reach their highest count after 36 h whereas cultures without the extract required 72 h to reach their peak counts, but the maximum cell count was higher for the extract-free culture. No biofilm was detected when P. aeruginosa was cultured in chicken broth at 10 °C when the extract was present. Addition of extract reduced proteolysis of actin, troponin-T and tropomyosin by P. aeruginosa, but had no effect on lipase activity. Therefore, AHL-CCE can be used to reduce the bacterial growth time and inhibit biofilm formation; thus, benefiting food safety and quality.     

Rheological Properties of Chitosan and its Interaction with Porcine Myofibrillar Proteins as Influenced by Chitosan's Degree of Deacetylation and Concentration

ABSTRACT Rheological and gelation characteristics of a specific degree of deacetylation (dd 50 to 99) and concentration (0.5% to 1.5%) of chitosan and chitosan/SSP (salt-soluble protein) mixtures were studied. Results suggested that SSP thermal gelation was interfered with by the positive charges of amino groups of the chitosan molecule. Regardless of degree of deacetylation, increasing chitosan concentration caused stronger interference on SSP gelation and lowered complex modulus [G*]. Loss tangent of chitosan/SSP gel decreased with increasing temperature. Higher level (1.5%) of dd 99 chitosan had the lowest G*, which resulted from ionic interaction with SSP. SDS-PAGE revealed protein aggregation, indicating possible covalent bond formation.     

不同工艺条件对大豆分离蛋白7S和11S组分影响的探讨

利用碱提酸沉的工艺，通过改变温度和离子强度得到不同参数条件下的大豆分离蛋白。经过凝胶电泳分析得到蛋白质的各个条带组分图，用图像捕捉成像技术以及相关软件，分析出分离蛋白各个组分的详细情况（包括组分数、分子量和各个组分所占的百分含量）。由实验可知，分离蛋白的7S和11S组分随温度和离子强度的改变而发生变化，分析其变化规律，从而确定合适的工艺参数。