绿豆和乳清蛋白水解物对双歧杆菌促生长条件的研究

对婴儿双歧(Bifidobacterium infantis)和两歧双歧杆菌(Bifidobacterium bifidum)的生长发育特性进行研究,在掌握双歧杆菌基本特性的基础上,以乳糖、乳清水解物和绿豆水解物为促生长因子,研究其对双歧杆菌的增殖影响.经过单因素和正交试验发现,乳清蛋白水解物和绿豆蛋白水解物对两种双歧杆菌都有显著的促生长作用.最佳培养基优化方案为:对婴儿双歧杆菌,在基础PTYG培养基基础上,添加1.5g/100mL的乳糖,40%的乳清蛋白水解物和40%的绿豆蛋白水解物;对两歧双歧杆菌,在基础PTYG培养基基础上添加0.5g/100mL的乳糖,40%的乳清蛋白水解物和40%的绿豆蛋白水解物.     

海藻酸钠和乳清蛋白作为益生菌包埋壁材的比较

利用海藻酸钠和乳清蛋白分别制备包埋有两歧双歧杆菌的微胶囊,测定了不同微胶囊的粒径、包埋效率、缓冲能力和外观形态,同时还考察了不同微胶囊对两歧双歧杆菌保护效果的影响。结果表明：乳清蛋白微胶囊的粒径和包埋效率均要高于海藻酸钠微胶囊,分别为202.5 μm,87.8%和118.3 μm,48.1%;虽然在高胆盐环境中两种微胶囊对两歧双歧杆菌的保护效果没有显著差别,但在低酸环境、模拟胃液和常温贮藏期中,相比于海藻酸钠微胶囊,乳清蛋白微胶囊将两歧双歧杆菌的存活量分别提高了大约5、2、0.5（lg（CFU/mL））。乳清蛋白微胶囊在pH值偏中性的环境中具有较高的缓冲能力;在外观形态上,由高浓度乳清蛋白溶液制备而来的微胶囊具有较好的呈球性和致密度,这些可能是乳清蛋白微胶囊具有较高保护效果的原因。     

两歧双歧杆菌BB-G90生产工艺的优化

对两歧双歧杆菌的4种发酵培养基进行了筛选,确定了适合两歧双歧杆菌BB-G90生长的发酵培养基;研究了5种冻干保护剂对两歧双歧杆菌BB-G90活菌的影响。利用筛选的发酵培养基培养BB-G90,在调控pH=5.0±0.5条件下进行300 L罐中试发酵试验,确定了发酵参数及冻干保护剂配方。试验结果表明:两歧双歧杆菌BB-G90在优化的发酵培养基、适宜的冻干保护剂及调控pH=5.0±0.5条件下发酵终止时间为22 h,此时,发酵液OD600值为5.62,发酵液活菌数为1.80×109CFU/mL,发酵液经离心、乳化及冻干后,菌粉活菌数为4.10×1011 CFU/g。     

中心组合设计优化两歧双歧杆菌BB01增殖培养基

为了提高两歧双歧杆菌培养液中的活菌数,采用中心组合设计对其增殖培养基进行了优化。在前期单因素试验的基础上,首先通过Plackett-Burman试验筛选出了影响两歧双歧杆菌生长的4个主因子,再通过中心组合设计试验及响应面分析确定了4个主因子的最佳浓度为:三氯化铁、水苏糖、蒺藜提取液和pH分别为0.012 g/L、8.0 g/L、2.5%和6.90,用优化后的增殖培养基培养两歧双歧杆菌,24 h后其培养液OD600为1.398±0.007,比优化前提高了24.28%。     

水苏糖、低聚木糖及半乳糖对双歧杆菌生长的影响

以MRS培养基为对照,研究了不同浓度的水苏糖、低聚木糖及低聚半乳糖对两歧双歧杆菌BB01生长的影响。结果表明:在MRS培养基中添加0.2%~1.0%的水苏糖对两歧双歧杆菌BB01的生长有一定的促进作用,添加0.2%~1.0%的低聚木糖对两歧双歧杆菌BB01的生长没有影响,添加0.2%~1.0%的低聚半乳糖对两歧双歧杆菌BB01生长有明显的促进作用,这可缩短培养时间。     

魔芋水解液对双歧杆菌生长促进作用

就魔芋水解液对双歧杆菌生长促进作用进行了体外观察实验,结果表明,在培养基中加入１％魔芋水解液,两歧双歧杆菌经３７℃培养１８ｈ后,活菌数量增殖了１７８．１０倍,而对照培养基中只增殖了２２．１４倍,活菌增殖量较对照提高了７．８５倍。     

双歧杆菌在发酵乳中活性的研究

分别使用两歧双歧杆菌和长双岐杆菌制作酸乳并冷藏21d,应用AMC,RMS培养基检测样品的双歧杆菌活菌数。发酵乳中的双歧杆菌的浓度达到10^8-10^9CFU/mL,在21d的冷藏中二种双歧杆菌的活菌数都超过了最小保健剂量10^6CFU/mL。RMS培养基同样有效地分离两歧双歧双歧杆菌和长双歧杆菌,可以采用涂布法和倾注法进行操作,RMS是对酸乳中双歧杆菌计数的最适宜的选择性培养基,使用AMC培养基由于涂布法造成较低的活菌计数。     

鲟鱼肝酶解条件优化及酶解液对双歧杆菌增殖效果的研究

为提高鲟鱼肝脏的加工附加值，使用碱性蛋白酶，以水解度为评价指标，在单因素试验的基础上，通过Plackett-Burman试验、响应面分析法优化鲟鱼肝蛋白酶解条件。在此条件下，进一步探究了酶解液对双歧杆菌的促生长效果。结果表明，最优酶解条件为碱性蛋白酶添加量10 540 U/g鱼肝蛋白、体系初始pH值10.0、酶解时间8.6 h、酶解温度45 ℃、料液比1∶10（g∶mL），此条件下水解度（81.7%）是优化前（40.4%）的1.02倍。鲟鱼肝蛋白酶解液对青春双歧杆菌、两歧双歧杆菌、动物双歧杆菌、长双歧杆菌、短双歧杆菌和婴儿双歧杆菌6种均具有显著的促生长效果。研究结果为鲟鱼加工副产物利用提供了新途径，同时也为双歧杆菌促生长因子的研发提供新思路。     

乳清蛋白在转谷氨酰胺酶作用下包埋益生菌的研究

为了提高双歧杆菌在人体胃肠道中的存活率,以乳清蛋白为壁材,转谷氨酰胺酶为交联剂,通过乳化凝胶的方法制备包埋有两歧双歧杆菌的蛋白质微球。实验表明:以此工艺制备的微球成球性较好,粒径为(308.2±16.2)μm,益生菌包埋率为87.8%±10.0%,与未包埋的两歧双歧杆菌比较,经过包埋后的两歧双歧杆菌在模拟胃液和高胆盐溶液中的存活率分别提高了5个和2个对数值。     

响应面法优化长双歧杆菌增殖培养基

为了提高长双歧杆菌发酵液中的活菌数,对其增殖培养基进行响应面优化。通过单因素试验筛选出长双歧杆菌的最佳碳源为乳糖,并发现低聚木糖、菊糖、低聚异麦芽糖、低聚果糖、苯丙氨酸、蛋氨酸、脯氨酸、谷氨酸及赖氨酸均能显著促进长双歧杆菌的生长。利用Design Expert 8.06软件设计Plackett-Burman 试验筛选出影响长双歧杆菌生长的3个最重要因子,通过Box-Behnken试验及响应面分析确定3个因子的最佳添加量为：低聚木糖1.7g/L、菊糖3.6g/L、脯氨酸0.4g/L,用优化后的增殖培养基培养长双歧杆菌,18h后其活菌数达(1.75±0.02)×109CFU/mL,比优化前提高了95.64%。     

Production of an exopolysaccharide-containing whey protein concentrate by fermentation of whey

Using whey as a fermentation medium presents the opportunity to create value-added products. Conditions were developed to partially hydrolyze whey proteins and then ferment partially hydrolyzed whey with Lactobacillus delbrueckii ssp. bulgaricus RR (RR; an EPS-producing bacterium). In preliminary experiments, pasteurized Cheddar cheese whey was treated with Flavourzyme to partially hydrolyze the protein (2 to 13% hydrolyzed). Fermentation (2 L, 38 degrees C, pH 5.0) with RR resulted in EPS levels ranging from 95 to 110 mg of EPS per liter of hydrolyzed whey. There were no significant differences in the amount of EPS produced during fermentations of whey hydrolyzed to varying degrees. Since a high level of hydrolysis was not necessary for increased EPS production, a low level of hydrolysis (2 to 4%) was selected for future work. In scale up experiments, whey was separated and pasteurized, then treated with Flavourzyme to hydrolyze 2 to 4% of the protein. Following protease inactivation, 60 L of partially hydrolyzed whey was fermented at 38 degrees C and pH 5.0. After fermentation, the broth was pasteurized, and bacterial cells were removed using a Sharples continuous centrifuge. The whey was then ultrafiltered and diafiltered to remove lactose and salts, freeze-dried, and milled to a powder. Unfermented hydrolyzed and unhydrolyzed whey controls were processed in the same manner. The EPS-WPC ingredients contained approximately 72% protein and 6% EPS, but they exhibited low protein solubility (65%, pH 7.0; 58%, pH 3.0).     

利用冷榨花生饼制备花生多肽饮料

以冷榨花生饼为原料,采用碱法和酶水解法制备花生蛋白,以蛋白质提取率为指标,确定蛋白提取条件,并利用所提取蛋白或蛋白水解物经乳酸菌发酵制备花生多肽饮料。结果表明NaOH溶液提取花生蛋白的最佳条件为：pH9.0、温度55℃、料液比1:8(g/mL)、浸提2h,蛋白提取率80.68%;胰蛋白酶水解蛋白的最佳条件为：酶与底物比1:50(m/m)、底物质量浓度5g/100mL、pH9.0、水解温度50℃,蛋白提取率96.26%。以花生水解蛋白和脱盐乳清粉为原料,采用直投式乳酸菌为发酵剂,发酵条件为：花生水解蛋白质量浓度2g/100mL、乳清粉加入量1g/100mL、发酵剂与发酵液比1:25(g/kg)、42℃发酵5h、4℃后熟15h、蔗糖质量分数9%时的口感最佳。     

Production of polysaccharide and surfactin by Bacillus subtilis ATCC 6633 using rehydrated whey powder as the fermentation medium

The aim of this research was to assess the amounts of polysaccharide and surfactin produced by Bacillus subtilis ATCC 6633 in rehydrated whey powder (RWP) as the growth medium. One-day-old cultures of B. subtilis (～4.6 log cfu/mL) were inoculated into 100mL of 10, 15, or 20% (wt/vol) RWP and incubated at 30°C for 72 h. To analyze the effects of lactose and protein on polysaccharide and surfactin production, 6 RWP solutions containing different levels of lactose and protein were also used as media. The number of vegetative cells and spores, pH, viscosity, and the concentration of lactose were determined at 0, 24, 48, or 72 h of fermentation. The levels of polysaccharide and surfactin produced after 72 h of fermentation were measured using HPLC and the phenol-sulfuric acid method, respectively. During 72 h of fermentation, B. subtilis populations increased from 4.6 to 10.54, 9.82, and 9.67 log(10) cfu/mL in 10, 15, and 20% RWP, respectively. The number of B. subtilis spores in 10% RWP increased from 3.91 to 4.72 log(10) cfu/mL after 48 and 72 h of fermentation, respectively. The increased level of lactose or protein in RWP did not significantly change the vegetative growth. After 72h of fermentation, the pH of RWP decreased from 5.70 to 4.99 with a slight increase in viscosity. Polysaccharide levels in 10, 15, and 20% RWP after fermentation were 513.6, 613.5, and 768.3mg/L, respectively, with B. subtilis producing 0.18 to 0.29 g/L of surfactin after 72 h of fermentation. The polysaccharide or surfactin production was not changed significantly by addition of protein or lactose to RWP. These results indicate that RWP is a good fermentation substrate for surfactin and polysaccharide production.     

Hydration of Whey Powders as Determined by Different Methods

Four methods for evaluating water hydration of 15 whey derivative powders were compared, and results are discussed with respect to the chemical composition of the powders. Hydration capacities between 0.21 and 4.64 mL water/g of powder were obtained, depending on the method used. The filtration/centrifugation method gave the highest hydration capacity, whereas the paste-water retention method gave the lowest. The Baumann test and the paste-water retention method were well correlated with protein and lactose content of the powders, enabling differentiation between hydration capacities of whey protein concentrates (35% proteins) and electrodialyzed whey powders (12% proteins). Reliable characterization of hydration required a combination of methods.     

Conversion of glucose in lactase-hydrolyzed whey permeate to fructose with immobilized glucose isomerase

The maximum conversion of glucose to fructose in lactase-hydrolyzed whey permeate by glucose isomerase was approximately 52% at .1 g enzyme/ml substrate after 7 h incubation at 60 degrees C. Removal of minerals from the substrate was essential for enzyme activity. The dependence of the enzyme on Mg++ and Co++ for activity in the presence of high ash concentration was demonstrated. Optimum Mg++ and Co++ additions were 250 and 100 ppm, respectively. The isomerization reaction was enhanced more when both 100 ppm Mg++ and 50 ppm Co++ were added. Hydrolyzed isomerized lactose whey syrup with sweetness equivalent to sucrose was successfully produced through enzymatic isomerization of glucose in lactase-hydrolyzed whey permeate after supplementation with pure glucose. Fructose in hydrolyzed isomerized lactose whey syrup was effectively separated from other sugars by Dowex 1X8-200 anion exchange resin in the bisulfite form.     

一种新型乳清碳酸饮料的制作工艺

介绍了一种将脱盐乳清粉配制成10%的水溶液,用β-半乳糖苷酶酶解降低其乳糖含量,经两组串联超滤装置处理得到澄清的饮料原液,最后调配加工成澄清型低乳糖碳酸饮料的制作工艺。结果表明,用以上处理方法可制得具有奶香风味、口感清爽并可供长期保存的乳清碳酸饮料。     

克鲁维酵母发酵乳清蛋白水解液生产乳清营养酒的研究

首先通过枯草芽胞杆菌水解乳清中的蛋白质,然后使用能够发酵乳糖的马克斯克鲁维酵母菌的驯化菌株发酵乳清蛋白水解液,经过离心除杂、浓缩、回流酒精、膜分离精致等工艺,制备富含可溶性蛋白肽的营养型低度乳清酒。     

酶法水解乳糖与热处理偶联对牛乳Maillard反应的影响

旨在探讨乳糖水解程度及热处理方法与Maillard反应的关系,鲜牛乳用中性乳糖酶处理获得不同水解程度的低乳糖牛乳,然后对牛乳进行不同的热处理,处理后的样本进行Maillard反应程度评价。采用葡萄糖氧化酶法测定不同水解时间的牛乳中葡萄糖质量浓度和乳糖水解率,用高效液相色谱法和紫外分光光度法分别测定水解后牛乳经不同热处理后的糠氨酸和5-羟甲基糠醛(5-hydroxymethylfurfural,5-HMF)含量及牛乳褐变程度的OD值。结果表明,随着乳糖水解时间的延长,牛乳中的葡萄糖含量呈增加的趋势,葡萄糖质量浓度从0.00 mg/100 m L增加到1 721.33 mg/100 m L,但增加趋势逐渐变缓;乳糖水解率从0%增加到70.33%,水解时间2.0 h后的牛乳水解率达到了50%以上。糠氨酸含量呈上升的趋势(P0.05),水解时间在3.0 h以上并经75℃、30 min热处理的牛乳,糠氨酸含量超过了190 mg/100 g pro;水解时间为0.5 h及以上并经75℃、15 s热处理的牛乳,糠氨酸含量超过了12 mg/100 g pro。生鲜牛乳和水解后经75℃、30 min热处理的牛乳,均未检测到5-HMF,水解后经75℃、15 s热处理的牛乳,随乳糖水解时间的延长,牛乳中5-HMF含量增加显著(P0.05)。牛乳的褐变程度随乳糖水解时间显著增加(P0.05),且乳糖酶水解后75℃、30 min热处理的牛乳的褐变程度明显高于75℃、15 s热处理的牛乳。本研究结果说明,乳糖经过酶水解后的牛乳,长时间热处理会加重乳Maillard反应,影响乳的蛋白质品质。     

Optimization of Thermal Pretreatment Conditions for the Separation of Native α-Lactalbumin from Whey Protein Concentrates by Means of Selective Denaturation of β-Lactoglobulin

ABSTRACT: In this study a method to obtain native α-lactalbumin with a high degree of purity of 98% (m/m) and recovery of 75% (m/m) by selective denaturation of β-lactoglobulin was developed. To achieve this goal, the thermal pretreatment of whey protein concentrate was optimized varying the composition of the liquid whey protein concentrate in terms of total protein, lactose and calcium content, and pH value. The kinetics of the thermal denaturation of α-la and β-lg were then investigated at predetermined optimal composition (protein content 5 to 20 g/L, lactose content 0.5 g/L, calcium content 0.55 g/L, and pH 7.5). Using the activation energies and reaction rate constants obtained, lines of equal effects for targeted denaturation degrees of α-la and β-lg were calculated. Depending on total protein content, an area of optimal heating temperature/time conditions was identified for each protein concentration level.