提取啤酒废酵母中葡聚糖及回收蛋白质工艺探讨

以啤酒废酵母为原料，采用较为简单、环保的酶促-自溶-碱法提取葡聚糖并回收蛋白质，结果表明，采用此法获得的葡聚糖得率和纯度分别为45．8％和33．6％；利用回收的蛋白质制备蛋白质饲料，得率达到91．49％。经成本核算，以年产10t葡聚糖计，每年可实现利润282．55万元，具有很好的发展前景。     

外加酶提高酵母自溶抽提物得率的研究

研究了外加中性蛋白酶、木瓜蛋白酶和β-葡聚糖酶对酵母自溶的影响。结果表明：在最适条件下，啤酒酵母和面包酵母的自溶干物质得率、蛋白质得率、氨基酸态氮得率分别提高52．2％、55，1％、14.6％和26.0％、32．8％、11．6％。     

超声-酶-碱法提取啤酒废酵母β-1，3-葡聚糖的研究

采用超声-酶-碱法从啤酒废酵母中提取β-1,3-葡聚糖,在超声波预处理和酶解最佳条件的同时,利用响应曲面法研究分析NaOH浓度、温度、用量和时间对β-1,3-葡聚糖得率、纯度和蛋白质含量的影响.试验结果表明,超声波处理后破壁率为94.22%;酶解后蛋白质去除率为62.82%;当加入2.05%的NaOH 30.50 mL,74℃处理5.7 h,β-1,3-葡聚糖的得率为10-21%,纯度为88.14%,蛋白质含量为1.19%.超声-酶-碱法处理工艺具有β-1,3-葡聚糖得率、纯度高、蛋白质含量低及提取时间短的特点.     

酶-碱法提取酵母β-1,3-葡聚糖的研究

采用酶-碱法从酵母自溶残渣中提取β-1,3-D-葡聚糖,通过正交试验得出最佳碱处理工艺条件为酶解后的沉淀物用60mL2%氢氧化钠溶液75℃处理6h,经冷冻干燥后,成品葡聚糖的得率为21.38%,其中多糖含量为92.17%,蛋白质含量为1.32%,水分含量为5.53%,酶-碱法处理工艺具有葡聚糖得率高、蛋白质含量低的特点。     

二次回归法优化燕麦β-葡聚糖提取工艺的研究

本文首先通过单因素实验，以β-葡聚糖的得率以及提取液中杂质的含量为指标，对水、强碱、弱碱这三种提取溶剂进行了选择，最终选择了以水作为提取溶剂。采用二次通用旋转回归实验对水提工艺进行了研究。研究结果得出最佳工艺条件为：pH10．0、温度70℃、提取时间60min，在此条件下β-葡聚糖的得率可达到8．47％。同时对去除淀粉、蛋白质、色素等关键步骤也进行了研究。     

酶处理纯化啤酒酵母β-1，3-葡聚糖的研究

采用酶处理对酵母残渣中β-1,3-葡聚糖进行纯化,研究了酶处理纯化的最佳工艺.结果表明:酵母残渣中添加208U/g底物的木瓜蛋白酶,在50℃、pH6.0条件下酶解8h,蛋白质去除率可达到62.82%,β-1.3-葡聚糖最终纯度为90.50%,得率为11.00%,经紫外光谱、薄层层析和性质分析为高纯度的β-1,3-葡聚糖,且回收到0.348g/L多肽、氨基酸含量丰富的蛋白水解液.     

大麦、麦芽中总氮、β-葡聚糖与玻璃质之间的某些联系

本文对与大麦和麦芽质量有关的总氮、β-葡聚糖和玻璃质含量之间的关系进行了研究。结果表明,三个大麦质量指标之间存在某些重要的联系。研究发现,大麦中β-葡聚糖和玻璃质百分含量之间存在负相关关系,与此相反的是玻璃质百分含量与氮含量之间存在着正相关关系。这些关系表明,当大麦中氮含量与产生游离氨基氮和多肽的蛋白质之间有关联时,玻璃质仅与发芽大麦中那些只产生游离氨基氮的蛋白质有关。尽管与氮有关的蛋白质在大麦发芽中会参与酶的形成,但与玻璃质有关的蛋白质并未在发芽中参与酶的形成。众所周知,“酶都是蛋白质,蛋白质却不都是酶”。在确定麦芽浸出物得率时,大麦中玻璃质和口．葡聚糖含量比总氮含量的作用更重要。     

双向发酵提取燕麦β-葡聚糖及其理化性质研究

从12种药用真菌中筛选出可增加燕麦β-葡聚糖含量的3种菌种--黄伞、大杯伞和灰树花,以我国的燕麦麸皮作为培养基,通过双向发酵法提取燕麦麸皮中的β-葡聚糖,进行工艺优化、分离纯化及理化性质的研究。结果表明3种菌经双向发酵后提取燕麦β-葡聚糖,其得率较未经发酵的燕麦β-葡聚糖得率高,分子量更小。与未经发酵的燕麦水提液相比,发酵液的蛋白质及总糖含量增加。     

超声波辅助提取青稞β-葡聚糖工艺优化

采用超声波辅助方法从青稞麸皮中提取β-葡聚糖。研究了水料比、pH、超声波功率、提取时间和提取温度对其得率的影响,通过正交试验优化了提取工艺。结果表明,超声波辅助提取青稞β-葡聚糖的最优工艺为:水料比1:18,提取温度45℃,超声波功率500 W,p H9,提取时间25 min,此条件下β-葡聚糖得率可达2.36%。各因素对β-葡聚糖得率的影响大小依次为:水料比>提取温度>超声波功率>pH>提取时间。比较了TCA法、Sevage法、木瓜蛋白酶法的除蛋白效果,木瓜蛋白酶法最佳,蛋白质去除率可达87.84%,β-葡聚糖保留率可达90.58%。     

酵母葡聚糖的制备及其理化性质研究

采用酶-碱法从酵母自溶残渣中提取β-1,3-D-葡聚糖,研究了提取工艺,并通过正交试验得出理想的酶处理工艺条件为:酶添加量为1600 IU/g,酶解3 h,pH8,温度60℃。碱处理工艺条件为:用60 mL 2%NaOH溶液在75℃处理酶解后的沉淀物6 h。冷冻干燥后得到成品葡聚糖,成品得率21.38%,其中多糖含量为92.17%,蛋白质含量为1.32%,水分含量为5.53%,该工艺具有得率高、蛋白质含量低的特点。应用红外光谱对糖分进行分析,成品为高纯度的酵母葡聚糖。     

Effect of dextrans on cryopreservation of goat cauda epididymal spermatozoa using a chemically defined medium

Experiments were carried out to investigate the cryoprotecting potential of dextrans (ranging from 10 to 2000 kDa) using a synthetic model system developed recently in this laboratory. Goat spermatozoa from the cauda epididymidis were extracted in a chemically defined medium (modified Ringer's solution) and assayed for motility using a phase-contrast microscope. The sperm cells were subjected to a freezing protocol in a computer-controlled biofreezer (cooling at 0.25 degrees C min(-1) to 5 degrees C; 5 degrees C min(-1) to -20 degrees C; 20 degrees C min(-1) to -100 degrees C) and plunged into liquid nitrogen. The frozen cells were thawed rapidly at 37 degrees C in a thermostatic waterbath. In the absence of dextran, all the spermatozoa lost their motility. The cryoprotecting efficacy of each dextran was found to be biphasic in nature. Initially, as the concentration of dextran was increased, the recovery of sperm motility also increased and reached an optimum value; however, with further increases in dextran concentration, the recovery of motility decreased sharply. Of all the sugar polymers tested, 10 kDa dextran showed the highest cryoprotecting efficacy, whereas the 2000 kDa sugar polymer was the least active. Dextrans of 10, 40, 73, 173, 252, 500 and 2000 kDa offered maximum cryorecovery of forward motility to the extent of approximately 23%, 21%, 19%, 18%, 16%, 15% and 8%, respectively. Optimum concentrations of these dextrans for cryoprotection of sperm motility were 8.42, 2.50, 1.09, 0.37, 0.31, 0.10 and 0.04 mmol l(-1), respectively. It thus appears that each dextran has a characteristic cryoprotection profile. The data show that the cryoprotecting efficacy and optimum cryoprotecting concentrations of dextrans are inversely related to their molecular mass. Dextran also served as a significant cryoprotectant in the presence of glycerol (0.87 mol l(-1)) and dimethyl sulphoxide (0.76 mol l(-1)), which are well known cryoprotectants; the action of the combined cryoprotectants was almost additive. The presence of glycerol or glycerol plus dimethyl sulphoxide caused a significant reduction (from 8.42 to 6.27 mmol l(-1)) in the optimum concentration of dextran. In the presence of the three cryoprotectants, recovery of sperm motility was as high as 58% (forward motility) and 60% (total motility).     

右旋糖酐/蚕豆蛋白复合凝胶的流变特性

为研究乳酸菌右旋糖酐对蚕豆蛋白食品相关性质的影响，采用哈克流变仪和质构仪等测定了添加不同浓度右旋糖酐时GDL诱导的酸致蚕豆蛋白复合凝胶质构和流变特性等指标。结果表明：添加右旋糖酐能显著增加蚕豆蛋白凝胶保水性，空白组蚕豆蛋白凝胶保水性为60.38%，1%右旋糖酐与蚕豆蛋白形成的复合凝胶保水性为70.08%（p<0.05）；右旋糖酐浓度在0.25%~1%之间，对复合蛋白凝胶色泽有一定影响；0.5%右旋糖酐与蚕豆蛋白形成的复合凝胶强度为0.27 N，与空白组蚕豆蛋白凝胶（0.37 N）差异显著，可软化含高蚕豆蛋白食品质构特性；右旋糖酐/蚕豆蛋白复合凝胶流动曲线符合carreau模型（R2>0.99），具有假塑性流体的特性；应变扫描的弹性模量G''均高于黏性模量G''，说明右旋糖酐/蚕豆蛋白复合凝胶的弹性占主导；频率扫描结果显示添加右旋糖酐可软化蚕豆蛋白凝胶，使凝胶G''、G''降低，更易于加工。在蚕豆食品中添加右旋糖酐可改善蚕豆蛋白的质构和流变特性，为拓展其应用范围提供参考。     

Improvement of functional properties of peanut protein isolate by conjugation with dextran through Maillard reaction

Reaction mixtures containing peanut protein isolate (PPI) and dextran (1:1 weight ratio) were dry-heated at 60 °C and 79% relative humidity for 7 days. SDS-PAGE analysis indicated that PPI had become complexed with dextran to form conjugates of higher molecular weight. Arachin, accounting for 50% of peanut proteins, was hard to glycosylate with dextran, which might limit the extent of glycosylation of PPI. The thermal stability of PPI was remarkably improved by mixture/conjugation with dextran. Proteins in mixtures/conjugates might have a more compacted tertiary conformation than PPI. The protein solubility of conjugates at pH 4.5-6.0 was remarkably increased compared with the PPI/mixture. Mixture with dextran could significantly improve the emulsifying and foaming properties of PPI (p < 0.05). Conjugation with dextran could further enhance emulsifying and foaming properties of PPI.     

酒精Haze改良法测定蔗汁α-葡聚糖含量

为了快速准确测定糖厂蔗汁中α-葡聚糖含量,对传统的酒精Haze法进行改良。利用淀粉酶与α-葡聚糖酶降解蔗汁中的淀粉与α-葡聚糖,以无淀粉与蔗汁α-葡聚糖的蔗汁作为空白制作α-葡聚糖在蔗汁中的工作曲线,测定样品α-葡聚糖含量。试验结果表明:该方法RSD为2.95%,加标回收率在94.2%～96.7%之间。与传统酒精Haze法测定结果相比准确度有了显著提高。该方法克服了其他检测方法的缺点,简化了实验步骤,加快了实验速度,为糖厂实时检测糖汁中的α-葡聚糖含量提供了新的途径。     

右旋糖苷修饰玉米SOD研究

为提高玉米SOD的稳定性,利用右旋糖苷对玉米SOD进行修饰.在不同条件下测定玉米SOD的活力,比较修饰前后玉米SOD稳定性.结果表明:修饰后玉米SOD的活性保持率为83%,修饰后玉米SOD的温度稳定性、pH稳定性均有显著提高,半衰期延长,对胃蛋白酶、胰蛋白酶耐受性增强.说明右旋糖苷是一种较好的玉米SOD化学修饰剂.     

葡聚糖延缓草鱼肌原纤维蛋白冷冻变性的机理分析

以草鱼肌原纤维蛋白为研究对象,通过研究盐溶性蛋白质量浓度、Ca2＋-ATPase活性、表面疏水性、总巯基和活性巯基含量在冷冻贮藏条件下的变化,比较不同分子质量葡聚糖（添加量0.5%）和传统商业抗冻剂（4%蔗糖＋4%山梨糖醇）对冻藏过程中肌原纤维蛋白的冷冻保护效果。结果表明,在冻藏过程中,葡聚糖能有效延缓草鱼肌原纤维蛋白的冷冻变性,效果优于传统商业抗冻剂;分子质量小的葡聚糖（T7）对Ca2＋-ATPase活性和活性巯基的保护作用要整体优于分子质量大的葡聚糖（T20）,冻藏超过15 d后延缓表面疏水性的上升和总巯基的保护上的效果也显著优于后者;因此,在工业中选用分子质量小的葡聚糖作为淡水鱼及相关制品的冷冻保护剂效果更佳。     

Formation of acid‐precipitated soy protein–dextran conjugates by Maillard reaction in liquid systems

The conjugation reaction between soybean acid‐precipitated protein (SAPP) and dextran in liquid systems via the initial stage of the Maillard reaction was studied. Functional SAPP–dextran conjugates were prepared in 80% ethanol‐reacting system at 50 °C for 6 h, along with 95% ethanol‐reacting system at 60 °C for 24 h. The covalent attachment of dextran to SAPP was confirmed by sodium dodecyl sulphate–polyacrylamide gel electrophoresis and gel filtration chromatography.Compared to the classical dry‐heating, the reaction time of glycosylation in the two ethanol systems was largely shortened. Emulsifying activity of SAPP–dextran conjugates obtained by dry‐heating incubation and in ethanol was similar at pH 7.0 and10.0, significantly higher than that of SAPP–dextran mixture or SAPP alone. In addition, SAPP–dextran conjugates obtained in 80% ethanol‐reacting system for 6 h were completely soluble after heating at 90 °C for 20 min. The impact of various processing conditions on the formation of SAPP–dextran conjugates was investigated. This study provides important guidance to create protein–polysaccharide conjugates at mild temperatures in liquid systems.     

Improvement of functional properties of acid-precipitated soy protein by the attachment of dextran through Maillard reaction

The functional acid-precipitated soy protein (SAPP)–dextran conjugate was prepared by dry-heated storage at 60 °C under 79% relative humidity (RH) for 5 days through Maillard reaction between the ε-amino of lysine in soy proteins and the reducing-end carbonyl residue in the dextran. The covalent attachment of dextran to SAPP was confirmed by SDS-polyacrylamide gel electrophoresis and gel filtration chromatography. Functional properties of soy protein depend on the structural and aggregation characteristics of their major components (storage globulins 7S and 11S). The conjugate seemed to be predominantly formed by 7S, and the acidic subunits of 11S in soy protein. The emulsifying properties of the SAPP–dextran conjugate were about four times higher than those of SAPP. The solubility of the protein was not enhanced as a result of preheating, but rather it was not decreased when the conjugated protein was heated at 90 °C for 20 min due to the presence of the polysaccharide. The excellent emulsifying properties of SAPP–dextran conjugate were maintained even at pH 3.0 and were further improved at pH 10.0. The object of Maillard reaction is to guarantee the suitable reaction degree, and the resulting soluble conjugate can have excellent emulsifying properties.     

Impact of dextran conjugation on physicochemical and gelling properties of sweet potato protein through Maillard reaction

Covalently linked sweet potato protein (SPP)–dextran conjugates were produced through Maillard reaction, and the effects of this conjugation on physicochemical and gelling properties of SPP were studied. Formation of SPP–dextran-conjugated product was confirmed with increase in browning intensity and molecular weight, with concomitant reduction in free amino group content of the protein. Dry-heating period of 0, 1, 3, 5 and 7 days gave rise to 0%, 12.8%, 19.0%, 25.2% and 22.8% degree of conjugation (DC), respectively. High molecular weight protein polymer with polydispersed band above 50 kDa was formed. FTIR spectroscopic analysis showed that the C-O (amide I) and C-N (amide II) stretching bands were modified by the Maillard reaction. Conjugation of SPP with dextran reduced its denaturation temperature. SPP–dextran conjugate gels’ network structures were sustained by both physical interactions and covalent bonds. SPP–dextran conjugates’ gels had significantly improved hardness and resilience in comparison with the unmodified SPP (P < 0.05).     

葡聚糖接枝对大豆蛋白功能特性及结构的影响

采用干热糖基化对大豆分离蛋白进行改性,研究其功能特质及结构特性。以葡聚糖和大豆分离蛋白(soy protein isolate,SPI)为原料,考察底物质量比和反应时间两个因素。结果表明:蛋白质与糖质量比2∶1,反应温度60℃时,产物接枝比较高,褐变程度中等;与SPI相比,糖基化之后大豆蛋白的溶解度提高了72.72%,乳化活性(emulsifying activity,EAI)和乳化稳定性(emulsion stability,ESI)分别提高了117.53%和134.20%。十二烷基硫酸钠聚丙烯酰胺凝胶电泳(sodium dodecyl sulfate-polyacrylamide gel electrophoresis,SDS-PAGE)表明SPI与葡聚糖发生了糖基化反应;傅里叶红外光谱(Fourier transform infrared spectroscopy,FT-IR)和荧光光谱分析表明,糖链的引入导致了大豆蛋白空间结构的变化;模拟体外消化特性结果表明,葡聚糖糖基化修饰对SPI体外消化性的改善效果不明显。