瑞士乳杆菌对契达干酪成熟期间所产ACE抑制肽的影响及其消化稳定性

为明确瑞士乳杆菌对契达干酪中血管紧张素转换酶（angiotensin-converting enzyme,ACE）抑制肽活性的影响,以蛋白质水解度和ACE抑制率为指标,与干酪乳杆菌组、鼠李糖乳杆菌组和空白组干酪进行对照,研究瑞士乳杆菌对干酪成熟期间蛋白质水解及ACE抑制活性的影响,并对ACE抑制活性最高时期的干酪进行消化稳定性研究。结果表明：成熟期间,3 组益生菌干酪的活菌数无明显差异（P＞0.05）,但均高于空白组;益生菌干酪的蛋白质水解程度和ACE抑制活性显著高于空白组（P＜0.05）,其中瑞士乳杆菌干酪的蛋白质水解程度最强,活性最高（79.71%）。模拟消化后,瑞士乳杆菌干酪活菌数降低14.30%,ACE抑制活性显著增加（P＜0.05）,达到86.06%,多肽质量浓度增加至2.81 mg/mL;研究不同分子质量超滤组分消化后的ACE抑制活性发现,其中大于10 kDa的多肽活性升高,小于10 kDa的活性下降。此外,添加瑞士乳杆菌不影响干酪的整体可接受性。因此,瑞士乳杆菌能促进干酪ACE抑制肽的产生并提高其活性,消化后活性的升高主要与大分子肽的降解有关。     

产ACE抑制肽菌株的筛选及其在模拟消化道环境中的活性分析

为了筛选出对血管紧张素转换酶（angiotensin-converting enzyme,ACE）抑制活性较高并且具有较强的耐酸耐胆盐和耐盐的乳酸菌,对实验室保存的27株乳酸菌进行了产酸能力、蛋白水解活力、ACE抑制活性、模拟胃肠道消化及酸、胆盐和盐耐受性的测定。结果显示,菌株M3的ACE抑制率可达71.94%±1.39%,具有较好的蛋白水解活力和产酸能力,蛋白水解活力为（86.66±3.51）μg/mL亮氨酸,滴定酸度为（71.67±2.86）°T。菌株M3经人工胃肠液分别处理3 h后,ACE抑制率为74.96%±1.73%;在pH3的环境中3 h,耐受性为14.34%±1.21%,活菌数能达到7 lg CFU mL?1以上;在含有0.3%胆盐的培养基中3 h,耐受性为37.50%±2.47%;在含有4% NaCl的培养基中24 h,耐受性为37.32%±1.84%。该菌经16S rDNA鉴定为Lactobacillus (Lb.) paracasei subsp. paracasei M3。因此,菌株Lb. paracasei subsp. paracasei M3可用作发酵牛乳富产ACE抑制肽且能耐受消化道环境具有益生菌潜力的菌株。     

钾盐替代对涂抹再制干酪的影响

以切达干酪为原料制成涂抹再制干酪产品,研究钾盐(KCl)替代对产品理化性质、质构性质、感官性质的影响。结果表明, KCl替代NaCl对干酪硬度和可涂抹性无显著影响(p0.05),感官测定中钾盐替代并没有呈现钾盐的苦味。KCl完全替代对感官性质无显著性影响(p0.05)且钠含量显著降低(p0.05),降低20.33%。因此,钾盐替代在涂抹再制干酪制作中具有一定可行性。     

乳酸菌胞外多糖发酵条件优化及抗肿瘤活性的研究

该研究以胞外多糖产量为评价指标,采用单因素试验及响应面试验对副干酪乳杆菌副干酪亚种（Lactobacillus paracasei subsp. paracasei）M5L产胞外多糖的发酵条件进行优化,并采用噻唑蓝（MTT）法及实时荧光定量聚合酶链式反应（RT-FQPCR）研究胞外多糖的抗肿瘤活性。结果表明,乳酸菌M5L产胞外多糖的最优发酵条件为发酵温度37 ℃、发酵时间12.8 h、初始pH值7.7、菌体浓度108 CFU/mL,在此最优发酵条件下,胞外多糖产量达到最大,为167.2 mg/mL,是优化前的1.3倍。当质量浓度为500 μg/mL的胞外多糖对Caco-2细胞处理72 h时,抑制作用最佳,抑制率达19.5%,显著增加胞内Caspase-3、Bax基因且降低Bcl-2基因的表达量,说明该胞外多糖可通过影响凋亡相关蛋白的表达来促进Caco-2细胞凋亡。     

副干酪乳杆菌TK1501发酵大豆粉对小鼠Ⅱ型糖尿病的干预作用

该实验旨在研究副干酪乳杆菌TK1501（Lactobacillus paracasei TK1501）发酵大豆粉对由链脲佐菌素（streptozotocin,STZ）诱导的Ⅱ型糖尿病模型小鼠的改善作用。取4周龄～5周龄SPF级昆明雄鼠,随机将其分为空白组、模型组、L.paracasei TK1501发酵大豆粉组、未发酵大豆粉组,每组各10只。连续腹腔注射STZ建立模型,连续给药4周。通过检测体重、血糖、糖耐量、肝糖原、胆固醇、甘油三酯和脏器组织病理学研究等,探究L.paracasei TK1501发酵大豆粉对STZ诱导小鼠Ⅱ型糖尿病的干预作用。结果显示,经L.paracasei TK1501发酵大豆粉干预的小鼠对葡萄糖耐受能力显著增强（p＜0.05）,小鼠的总胆固醇（p＜0.01）、低密度脂蛋白胆固醇（p＜0.01）、高密度脂蛋白胆固醇（p＜0.01）和糖原（p＜0.05）等指标得到显著改善。同时,患病小鼠的肝脏、肾脏和胰腺受损得到明显改善。     

副干酪乳杆菌对酒精致肝损伤小鼠生长性能和肝脏的影响

该实验旨在研究副干酪乳杆菌（Lactobacillus paracasei）M5L对酒精致肝损伤小鼠生长性能及肝功能的影响。试验选取40只生长性能良好、体质量在25～28 g之间的昆明小鼠，随机将小鼠分为对照组、模型组、高、低副干酪乳杆菌剂量组，每组各10只，饲养28 d后进行生长性能和肝功能指标的测定。结果表明，与模型组的小鼠相比，副干酪乳杆菌M5L处理后能显著提高小鼠的体质量（P＜0.05），显著增大胸腺指数和脾脏指数（P＜0.05）；减轻小鼠肝脏水肿、充血，肝细胞坏死的情况，并且显著降低小鼠血清中谷丙转氨酶（ALT）、天门冬氨酸氨基转移酶（AST）活性（P＜0.05），显著降低小鼠肝脏中丙二醛（MDA）的含量（P＜0.05），显著增加小鼠肝脏中超氧化歧化酶（SOD）活性和谷胱甘肽（GSH）的含量（P＜0.05）。综上，副干酪乳杆菌M5L能够提高酒精致肝损伤小鼠的生长性能，并增强小鼠肝脏的抗氧化能力。     

益生菌对契达干酪成熟过程中抗氧化性变化的影响

为确定益生菌对契达干酪抗氧化性变化的影响,在菌株具备良好耐酸、耐盐性,适用于干酪生产前提下,以水解性和抗氧化性为指标,分别筛选出水解能力和抗氧化能力较强的菌株,并将其添加到契达干酪中,不添加益生菌的干酪为空白组,对干酪成熟过程中活菌数和抗氧化性进行分析。结果表明,9?株益生菌中,瑞士乳杆菌（Lactobacillus helveticus）1.0612和鼠李糖乳杆菌（Lactobacillus rhamnosus）1.0911分别具有较强的水解能力和抗氧化能力。在成熟过程中,添加L. helveticus 1.0612和L. rhamnosus 1.0911的两组干酪活菌数无显著差异,但均显著高于空白组。3?组干酪抗氧化能力均先升高再降低、最后趋于平缓,1,1-二苯基-2-三硝基苯肼（1,1-diphenyl-2-picrylhydrazyl,DPPH）自由基和羟自由基清除能力均在第4个月达到最大,还原能力在第5个月达到最大,且添加水解能力强的L. helveticus 1.0612干酪各项抗氧化能力的最大值（DPPH自由基、羟自由基清除能力和还原能力分别为51.05%、49.97%、0.66）均显著高于添加L. rhamnosus 1.0911的干酪（47.30%、46.19%、0.56）（P＜0.05）。因此,在契达干酪中添加水解能力较强的菌株,相比于添加本身具有良好抗氧化活性的菌株,可能会加剧干酪的蛋白水解,生成具有抗氧化能力的短肽和氨基酸,从而提高干酪的抗氧化活性。     

瑞士乳杆菌H11发酵乳饮料的ACE抑制活性及代谢组学研究

目的:研究瑞士乳杆菌H11与副干酪乳杆菌Lc-01两种发酵乳饮料在贮藏期间的代谢差异变化。方法:使用气相色谱-质谱（SPME-GC-MS）联用、高效液相色谱（HPLC）和超高效液相色谱串联四级杆飞行时间质谱（UPLC/Q-TOF MS）技术对4 ℃、贮藏28 d期间发酵乳饮料中的挥发性风味物质、代谢物以及ACE抑制活性之间的差异进行分析。结果:在4 ℃贮藏28 d后,瑞士乳杆菌H11发酵乳饮料的体外ACE抑制活性比副干酪乳杆菌Lc-01高60%以上,ACE抑制肽VPP和IPP含量也显著高于Lc-01（P<0.05）。采用SPME-GC-MS发现瑞士乳杆菌H11发酵乳饮料中香气成分丰富,特征风味物质2-庚酮和2-壬酮相对含量较高,分别为43.84%和12.39%。基于UPLC/Q-TOF MS的结果表明,贮藏期间两种发酵乳饮料的主要代谢差异物为肽、氨基酸和有机酸。结论:瑞士乳杆菌H11在制备发酵乳饮料方面存在巨大潜力。     

副干酪乳杆菌诱导Caco-2细胞凋亡及其发酵乳贮藏品质的研究

以新疆马奶酒中自行分离得到的副干酪乳杆菌（Lactobacillus paracasei）M5L为对象,研究该菌株对结肠癌Caco-2细胞的抑制作用并分析发酵乳贮藏品质变化。结果表明,副干酪乳杆菌M5L可以在一定的盐浓度、胆盐浓度及酸性环境中较好的生长。当菌体以100∶1的感染系数作用于Caco-2细胞72 h后抑制率最佳;显微镜下观察活菌处理后的细胞发生明显的凋亡现象;流式细胞仪检测到副干酪乳杆菌M5L可以诱导Caco-2细胞内活性氧显著增加,并将细胞周期阻滞在合成期,且通过增大Caspase-3和Bax基因表达量,降低Bcl-2基因表达量来促进肿瘤细胞的凋亡。其发酵酸乳贮藏5 d时品质较佳,长时间的贮存会导致酸乳品质的劣变。     

大豆多肽的降压活性及其相对分子质量分布研究

采用以FAPGG为底物的酶活力检测法对由不同蛋白酶（碱性蛋白酶、胰蛋白酶、风味蛋白酶、木瓜蛋白酶和菠萝蛋白酶）水解制备的大豆多肽的降压活性进行了测定。结果表明,碱性蛋白酶、胰蛋白酶和风味蛋白酶三酶联合水解制备的大豆多肽具有较好的降压活性,其ACE抑制率达62.78%。经超滤发现,5kDa膜滤过液中多肽含量最多,占总量的73.44%,降压活性最强,其ACE抑制率达84.44%。最后,采用SephadexG-25凝胶色谱进一步分离纯化,得到5个峰,其相对分子质量在1 200以下的组分约占75%,其中相对分子质量在250—600左右的组分占47.6%,降压活性最强,ACE抑制率达92.15%。     

Microbiological study of Manura, a hard cheese made from raw ovine milk in the Greek island Sifnos

Changes in the microbial flora of Manura, a raw ovine milk cheese, were studied during ripening. In general, the various microbial groups developed better on the cheese surface than in the interior, but red wine treatment had an inhibitory effect on their growth and microbial counts decreased ( P  < 0.05) more rapidly on the cheese surface than in the interior. NaCl and moisture of the cheese affected microbial levels significantly. Thus, Enterobacteriaceae and coliforms were reduced sharply ( P  < 0.05) during ripening on a straw bed (∼3 months) and they were not detected in mature cheese. Lactic acid bacteria predominated over the other microbial groups throughout ripening. Leuconostoc mesenteroides ssp. cremoris , Pediococcus pentosaceus and Lactobacillus paracasei ssp. paracasei , frequently found in maturing cheese, could be used as starters to make this cheese. Moreover, the lactic acid bacteria predominating in mature cheese, such as Weissella paramesenteroides , Lactobacillus bifermentans and Lactobacillus brevis , may contribute to cheese ripening through their biochemical activities.     

Proteolysis of low-moisture Mozzarella cheese as affected by substitution of NaCl with KCl

The proteolytic and ACE inhibitory activities of low-moisture Mozzarella cheese (LMMC) as affected by partial substitution of NaCl with KCl were investigated. Experimental LMMC were made and salted with 4 salt mixtures: NaCl only (control), 3NaCl:1KCl, 1NaCl:1KCl, and 1NaCl:3KCl, and then proteolytic activity and angiotensin-converting enzyme inhibitory activity were determined. Salt treatment significantly affected angiotensin-converting enzyme inhibitory activity and phosphotungstic acid-soluble N of LMMC during storage. Water-soluble N, trichloroacetic acid-soluble N, lactic acid bacteria population, and total free amino acids were unaffected during storage. Nonetheless, water-soluble N and trichloroacetic acid-soluble N increased significantly during storage within a salt treatment. Peptide profiles and urea-PAGE gels did not differ between experimental cheeses at the same storage time.     

Effect of lactobacillus adjunct cultures on the microbiological and physicochemical characteristics of Roncal-type ewes'-milk cheese

The effect of two different experimental adjunct cultures composed of native facultatively heterofermentative lactobacilli (FHL) on the development of various groups of micro-organisms in Roncal-type ewes' milk cheese was studied. Four cheese batches were manufactured from raw milk (C), pasteurized milk (P), pasteurized milk and an adjunct culture of Lactobacillus paracasei (PP); and pasteurized milk and adjunct culture of Lactobacillus paracasei plus Lactobacillus plantarum (PPP). Retention of the two adjunct cultures in the cheeses was good, and population levels remained constant at around 10(7) cfu g(-1) of cheese throughout ripening. Levels of Enterobacteriaceae and enterococci fell off more abruptly in the batches made with the Lactobacillus adjunct cultures, suggesting competition between the added lactobacilli and those groups of micro-organisms. The inhibitory effect was greater for the adjunct culture composed of L. paracasei plus L. plantarum. Lactococcal levels were higher in the batches made with added FHL, which may be indicative of a synergistic effect between these two groups.     

Salt Reduction in a Model High‐Salt Akawi Cheese: Effects on Bacterial Activity,pH, Moisture,Potential Bioactive Peptides,Amino Acids,and Growth of Human Colon Cells

This study evaluated the effects of sodium chloride reduction and its substitution with potassium chloride on Akawi cheese during storage for 30 d at 4 °C. Survival of probiotic bacteria (Lactobacillus acidophilus, Lactobacillus casei, and Bifidobacterium longum) and starter bacteria (Streptococcus thermophilus and Lactobacillus delbrueckii ssp. bulgaricus), angiotensin‐converting enzyme‐inhibitory and antioxidant activities, and concentrations of standard amino acids as affected by storage in different brine solutions (10% NaCl, 7.5% NaCl, 7.5% NaCl+KCl [1:1], 5% NaCl, and 5% NaCl+KCl [1:1]) were investigated. Furthermore, viability of human colon cells and human colon cancer cells as affected by the extract showing improved peptide profiles, highest release of amino acids and antioxidant activity (that is, from cheese brined in 7.5% NaCl+KCl) was evaluated. Significant increase was observed in survival of probiotic bacteria in cheeses with low salt after 30 d. Calcium content decreased slightly during storage in all cheeses brined in various solutions. Further, no significant changes were observed in ACE‐inhibitory activity and antioxidant activity of cheeses during storage. Interestingly, concentrations of 4 essential amino acids (phenylalanine, tryptophan, valine, and leucine) increased significantly during storage in brine solutions containing 7.5% total salt. Low concentration of cheese extract (100 μg/mL) significantly improved the growth of normal human colon cells, and reduced the growth of human colon cancer cells. Overall, the study revealed that cheese extracts from reduced‐NaCl brine improved the growth of human colon cells, and the release of essential amino acids, but did not affect the activities of potential bioactive peptides.     

CONSUMER EVALUATION OF SODIUM REDUCED, RESTRUCTURED LAMB ROASTS

The longissimus dorsi (LD) and psoas major (PM) muscles were removed from the racks and loins of nine lamb carcasses (18 sides). Each LD was cut in half to yield 36 pieces of each muscle. Three defatted LD pieces and one PM were randomly grouped together (4 replications) and assigned to the following treatments at 1.5% of the fresh muscle weight: (1) 100% NaCl (control); (2) 75% NaCl + 25% KCl; (3) 50% NaCl + 50% KCl. Water (5%) and sodium tripolyphosphate (.3%) were also added. Muscle pieces were tumbled for 2 h, tied into roasts, cooked and smoked. Proximate analysis results showed no significant differences (P > 0.05) among treatment groups for percentage moisture and fat. The 100% NaCl control had significantly higher (P < 0.05) cooking yields than the 50% KCl replacement treatment. A consumer sensory panel found no consistent differences among treatments for juiciness, flavor desirability, off-flavor, or overall acceptability among treatments. Consumer panel responses indicate that the low-fat (5.3–6.8%), low-sodium (455–744 mg/100g), restructured lamb roasts evaluated in this experiment would be an acceptable new processed lamb product.     

The effect of sodium reduction with and without potassium chloride on the survival of Listeria monocytogenes in Cheddar cheese

Sodium chloride (NaCl) in cheese contributes to flavor and texture directly and by its effect on microbial and enzymatic activity. The salt-to-moisture ratio (S/M) is used to gauge if conditions for producing good-quality cheese have been met. Reductions in salt that deviate from the ideal S/M range could result in changing culture acidification profiles during cheese making. Lactococcus lactis ssp. lactis or Lc. lactis ssp. cremoris are both used as cultures in Cheddar cheese manufacture, but Lc. lactis ssp. lactis has a higher salt and pH tolerance than Lc. lactis ssp. cremoris. Both salt and pH are used to control growth and survival of Listeria monocytogenes and salts such as KCl are commonly used to replace the effects of NaCl in food when NaCl is reduced. The objectives of this project were to determine the effects of sodium reduction, KCl use, and the subspecies of Lc. lactis used on L. monocytogenes survival in stirred-curd Cheddar cheese. Cheese was manufactured with either Lc. lactis ssp. lactis or Lc. lactis ssp. cremoris. At the salting step, curd was divided and salted with a concentration targeted to produce a final cheese with 600 mg of sodium/100 g (control), 25% reduced sodium (450 mg of sodium/100 g; both with and without KCl), and low sodium (53% sodium reduction or 280 mg of sodium/100 g; both with and without KCl). Potassium chloride was added on a molar equivalent to the NaCl it replaced to maintain an equivalent S/M. Cheese was inoculated with a 5-strain cocktail of L. monocytogenes at different times during aging to simulate postprocessing contamination, and counts were monitored over 27 or 50 d, depending on incubation temperature (12 or 5°C, respectively). In cheese inoculated with 4 log₁₀ cfu of L. monocytogenes/g 2 wk after manufacture, viable counts declined by more than 3 log₁₀ cfu/g in all treatments over 60 d. When inoculated with 5 log₁₀ cfu/g at 3 mo of cheese age, L. monocytogenes counts in Cheddar cheese were also reduced during storage, but by less than 1.5 log₁₀ cfu/g after 50 d. However, cheese with a 50% reduction in sodium without KCl had higher counts than full-sodium cheese at the end of 50 d of incubation at 4°C when inoculated at 3 mo. When inoculated at 8 mo postmanufacture, this trend was only observed in 50% reduced sodium with KCl, for cheese manufactured with both cultures. This enhanced survival for 50% reduced-sodium cheese was not seen when a higher incubation temperature (12°C) was used when cheese was inoculated at 3 mo of age and monitored for 27 d (no difference in treatments was observed at this incubation temperature). In the event of postprocessing contamination during later stages of ripening, L. monocytogenes was capable of survival in Cheddar cheese regardless of which culture was used, whether or not sodium had been reduced by as much as 50% from standard concentrations, or if KCl had been added to maintain the effective S/M of full-sodium Cheddar cheese.     

Biopreservation by Lactobacillus paracasei in coculture with Streptococcus thermophilus in potentially probiotic and synbiotic fresh cream cheeses

The viability of Lactobacillus paracasei and its effect on growth of the microbiota in potentially probiotic and synbiotic fresh cheeses during storage at 4 +/- 1 degree C was investigated. Three cheese-making trials (T1, T2, and T3) were prepared in quadruplicate, all supplemented with a Streptococcus thermophilus culture. L. paracasei subsp. paracasei was added to cheeses in T1 and T2, and inulin was added to cheeses in T2. Counts of L. paracasei, S. thermophilus, coliforms, Escherichia coli, Staphylococcus spp., DNase-positive Staphylococcus, and yeasts and molds were monitored during storage for up to 21 days. Viable counts of L. paracasei in probiotic (T1) and synbiotic (T2) cheeses remained above 7 log CFU/g during the entire storage period, whereas counts of S. thermophilus remained above 9.5 log CFU/g for cheeses from TI, T2, and T3. Populations of coliforms, Staphylococcus spp., and DNase-positive Staphylococcus were higher in T3 cheese and differed significantly from those in cheeses from T1 and T2 (P < 0.05). Inhibition of contaminants prevailed when both L. paracasei and S. thermophilus were present in fresh cream cheese and probably was due to acid production by both strains; bacteriocin production was not found. Addition of inulin in T2 did not impact microbial viability (P > 0.05). L. paracasei subsp. paracasei in coculture with S. thermophilus was inhibitory against microbial contaminants in fresh cream cheese with or without the addition of inulin, indicating the potential use of this combination in a probiotic and synbiotic product.     

基于人体必需金属盐替代的低钠盐肌原纤维蛋白凝胶特性和作用机理

为降低肌原纤维蛋白凝胶中钠盐的使用量,研究不同氯盐(CaCl₂、MgCl₂、KCl)部分替代NaCl对肌原纤维蛋白凝胶性质包括强度、微观结构、持水力和流变学性质的影响,并通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳、内源性荧光光谱、表面疏水性、巯基含量等技术手段阐明其作用机理。结果表明,3种氯盐替代效果排序为KCl>CaCl₂>MgCl₂。0.5%～1.5%质量分数的盐替代范围内,KCl替代组的凝胶强度优于对照组(3%NaCl),持水力与对照组相比无显著差异(P>0.05);Ca Cl₂替代组的凝胶强度在1.5%替代质量分数时显著下降(P<0.05),持水力呈先下降后上升趋势;MgCl₂替代组凝胶强度显著降低(P<0.05),持水力上升。在流变学性质方面,不同质量分数KCl替代组的储能模量明显高于对照组。CaCl₂和MgCl₂的替代使肌原纤维蛋白表面疏水性增大、巯基含量减少,3种氯盐的替代...