植物乳杆菌SP-3对干酪挥发性风味物质的影响

采用固相微萃取(SPME)富集干酪模型中的挥发性风味物质,并以气相色谱—质谱联用仪(GC-MS)检测结果来评定植物乳杆菌SP-3对干酪挥发性风味的影响。结果表明,添加植物乳杆菌SP-3的干酪模型中2-丁酮、2-甲基-1-丙醇、3-甲基-1-丁醇、1-戊醇、4-甲基-2-戊醇、3-羟基-2-丁酮等具有坚果味、水果味和奶油味的物质明显高于对照组,而二甲基硫化物和3-甲硫基-1-丙醇的浓度则低于对照组。     

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。     

辅助发酵剂加速Provolone干酪成熟的研究

将瑞士乳杆菌6024作为辅助发酵剂添加到Provolone干酪中,加速Provolone干酪成熟,研究了其对干酪游离氨基酸、游离脂肪酸、质构特性、电镜、风味物质的影响。结果表明,瑞士乳杆菌6024作为辅助发酵剂对干酪中游离脂肪酸质量分数没有显著影响,但能显著增加干酪游离氨基酸质量分数,60 d时达到对照组90 d时的质量分数,此时干酪的质构特性、微观结构、风味物质与对照组90 d时差异不显著。由此可知,瑞士乳杆菌6024作为辅助发酵剂可加速Provolone干酪的成熟,缩短成熟时间。     

植物乳杆菌S72对高达干酪品质的影响

以实验室自主分离的一株具有优良功能特性的植物乳杆菌(Lactobacillus plantarum) S72为辅助发酵剂,制备高达干酪(Gouda),研究植物乳杆菌S72对干酪理化指标、质构、活菌数、游离氨基酸及游离脂肪酸等的影响。结果表明:与商品发酵剂组相比,添加L.plantarum S72作为辅助发酵剂的Gouda干酪,在干酪成熟末期,L. plantarum S72保持较高的活菌数,对干酪的理化指标及质构特性无不良影响。L. plantarum S72提高了Gouda干酪成熟过程中WSE/TN和70%Ethanol-SN的含量,显著增加了PTA-SN的含量。L.plantarum S72后有助于Gouda干酪成熟过程中游离氨基酸的生成。这些结果说明添加L. plantarum S72作为辅助发酵剂不会影响Gouda干酪的质地,并且对Gouda干酪成熟过程中的风味有促进作用。     

对添加附属发酵剂干酪的感官分析

对添加附属发酵剂SP-3的干酪进行感官评定,结果表明,筛选得到的植物乳杆菌SP-3作为附属发酵剂,对半硬质干酪的风味产生影响.同时,对照组干酪的主要风味特征为易于接受的坚果风味和奶香风味.     

凝乳酶添加量对牦牛乳硬质干酪蛋白质降解的影响

通过测定不同可溶性氮含量以及采用尿素凝胶电泳法,研究了不同小牛皱胃酶添加量对牦牛乳硬质干酪3个月成熟过程中蛋白质降解的影响,并对干酪苦味进行了感官评价。研究表明:凝乳酶添加量对干酪p H 4.6SN和12%TCASN影响显著(P0.05),在成熟1个月时,不同凝乳酶添加量干酪p H 4.6SN之间差距较大,且凝乳酶添加量与干酪p H 4.6SN间存在较强线性关系。凝乳酶添加量对5%PTASN和游离氨基酸影响不显著(P0.05)。尿素凝胶电泳显示:干酪中αs-酪蛋白降解依赖于凝乳酶添加量,且降解程度大于β-酪蛋白。凝乳酶添加量对干酪苦味影响显著(P0.05),且随着凝乳酶添加量的增多,其苦味程度逐渐加重,但是大部分干酪苦味属于轻微苦味和中等程度苦味之间。干酪苦味与12%TCASN、αs-酪蛋白和β-酪蛋白降解率之间具有较强相关性(Spearman相关系数0.7,P0.01)。     

乳酸菌自溶对切达干酪成熟中蛋白质分解的影响

选取自溶度不同的乳杆菌作为附属发酵剂混合商业发酵剂制作切达干酪,研究了乳酸菌自溶对干酪成熟的影响.测定成熟期间各干酪中乳酸菌活菌数、pH值、pH4.6可溶氮质量分数和12%TCA可溶氮质量分数.并结合SDS-PAGE分析干酪蛋白质的水解情况.结果表明,各实验组干酪的12%TCA可溶性氮含量随着成熟时间延长逐渐增加.在1个月后不同组别之间差异显著(P<0.05).与对照组干酪相比,加入高自溶度菌株的实验组干酪蛋白分解程度较高,非蛋白氮的质量分数为8.00%.同时,SDS-PAGE结果显示,各干酪的蛋白质都有-定程度的分解.产物略有不同.乳酸菌自溶可以加速蛋白质分解.     

协同发酵在植物乳杆菌发酵乳中的应用研究进展

植物乳杆菌由于其益生功能受到广泛关注。植物乳杆菌蛋白水解系统不完全,存在肽转运系统和胞内肽酶,但缺乏胞壁蛋白酶,这导致该菌无法直接利用乳中蛋白质,因此在乳中生长不佳。利用蛋白酶活性强的乳酸菌和植物乳杆菌的协同发酵是一种解决方法,前者能初步水解乳中蛋白质产生多肽和游离氨基酸,为植物乳杆菌提供氮源,促进其生长。本文综述了植物乳杆菌益生功能,在乳中生长不良的原因及相关解决办法(添加营养物质、添加蛋白酶、协同发酵)的研究进展。     

用于低脂干酪开发的附属发酵剂的筛选

目的筛选出生产低脂干酪所需添加较为适宜的附属发酵剂。方法选择7株乳酸菌作为研究对象,测定其总肽酶活力、菌株生产能力、产酸能力、产粘能力、菌体自溶度、产胞外多糖数量、蛋白质水解能力等7种指标。结果 7株乳酸菌中嗜酸乳杆菌(Lactobacillus acidophilus,LA)总肽酶活力最高,为(256.46±11.88)μg/L,植物乳杆菌(Lactobacillus plantarum,LP)最低;菌株生长能力LA最好;菌株副干酪乳杆菌(Lactobacillus paracasei,LPC)在发酵10 h后的产酸能力最强且趋于稳定,pH在3.8左右;LP菌株产粘能力最优,为(168.10±14.72)mPa·s;瑞士乳杆菌(Lactobacillus helveticus,LH)菌株自溶度最好,达到35%;产胞外多糖数量最优菌株是LP,含量为(792.69±35.94)mg/L;LH的蛋白质水解能力最高,水解产生的游离氨基酸含量为(94.78±2.82)mg/L。结论 LH作为低脂干酪的附属发酵剂较好。     

瑞士乳杆菌对Gouda干酪品质的影响

将瑞士乳杆菌作为辅助发酵剂添加到Gouda干酪中,以未添加瑞士乳杆菌的干酪作为空白对照,观察Gouda干酪在成熟期间的成分、成熟率、蛋白质降解、组织状态的变化情况。同时通过测定氨基酸含量及品尝试验对干酪进行感官评价。结果显示:在120 d的成熟期内两种干酪的蛋白质、脂肪、水分、盐分和灰分含量均未发现明显的差异,而添加瑞士乳杆菌的干酪成熟第30天的成熟率、SDS-PAGE电泳条带数量以及谷氨酸含量均明显高于对照样。添加0.1%的瑞士乳杆菌不仅加快Gouda干酪的成熟,而且提高了干酪的感官品质。     

海藻酸钠固定化乳酸菌促熟干酪效果的研究

对海藻酸钠固定化乳酸菌促熟干酪的效果进行了研究。结果表明，在干酪粒中添加10^6CFU／g固定化乳酸菌，水溶性氮（WSN）、三氯乙酸氮（TCASN）和磷钨酸氮（PTASN）含量较对照组明显增大；ADV值在添加固定化乳酸菌干酪中成熟初期变化不大，45d后明显增大；感官评定结果表明，干酪粒中添加10^5CFU／g固定化乳酸菌干酪成熟60d时风味和质地较好，可比对照组干酪成熟期缩短30d左右。     

Influence of ripening on proteolysis and lipolysis of Murcia al Vino cheese

The aim of this study was to evaluate the influence of five different manufacturers and two ripening periods on the proteolysis and lipolysis patterns of Murcia al Vino goat cheese. The manufacturers significantly affected the water activity (a_w), pH, dry matter and fat content, several nitrogen fractions: water soluble nitrogen (WSN), trichloroacetic acid (12% w/v) soluble nitrogen (TCASN) and phosphotungstic acid (5% w/v) soluble nitrogen (PTASN); also the free amino acid (FAA) and free fatty acid (FFA) contents, with the exception of C_4:0, C_16:0 and C_18:0. Different ripening periods significantly affected the dry matter content, WSN and PTASN and all FAA, except serine.     

Proteolysis in goat cheese made from raw,pasteurized or pressure-treated milk

Primary and secondary proteolysis of goat cheese made from raw (RA), pasteurized (PA; 72 °C, 15 s) and pressure-treated milk (PR; 500 MPa, 15 min, 20 °C) were examined by capillary electrophoresis, nitrogen fractionation and HPLC peptide profiles. PA milk cheese showed a more important hydrolysis (P<0.05) of α_s1-casein than RA milk cheese at the first stages of ripening (15 days), while PR milk cheese had a level between those seen in PA and RA milk cheeses. Degradation of β-casein was more important (P<0.05) in PA and PR than in RA milk cheeses at 15 days of ripening. However, from thereon β-casein in PR and RA milk cheeses was hydrolyzed at essentially similar rates, but at lower rates (P<0.05) than in PA milk cheeses. Pressure treatment could induce proteolysis of β-casein in a way, which is different from that produced by heat treatment. There was an increase in 4.6-soluble nitrogen (WSN) and in trichloroacetic acid (TCASN) throughout ripening in cheeses, but higher contents (P<0.05) in PA and PR milk cheeses at the end of ripening were observed. PR milk cheeses contained considerably higher content (P<0.05) of free amino acids than RA or PA milk cheeses. In general, heat and pressure treatments had no significant effect on the levels of hydrophobic and hydrophilic peptides.     

Effects of Commercial Food Grade Enzymes on Proteolysis and Textural Changes in Granular Cheddar Cheese

Effects of some commercial food grade enzymes on the proteolysis and textural changes in granular cheese were studied. A neutral protease and lipase/protease combination-treated cheese samples showed significantly higher proteolysis than control samples at two ripening temperatures. However, lipase treatment alone had little effect on proteolysis. Cheese with a culture/enzyme mixture showed faster production of total free amino acids, with gross proteolysis similar to that of controls. Various non-linear correlations were observed between total amino acid production and gross proteolysis. Statistical analysis indicated that gross proteolysis measurements may directly reflect texture development as well as levels of proteolysis, whereas the total free amino acids relates less to the texture development. Multiple regression equations were developed for the relationship.     

Amino acid and soluble nitrogen evolution throughout ripening of Serra da Estrela cheese

Four batches of Serra da Estrela cheese originating from as many dairy farms were sampled throughout the ripening period, and assayed for the evolution of free amino acid (FAA) content, total nitrogen content (TN), water-soluble nitrogen content (WSN), trichloroacetic acid-soluble nitrogen content (TCASN) and phosphotungstic acid-soluble nitrogen content (PTASN). The WSN content increased from 1% (on the day of manufacture) up to 43% of TN by 180 d of ripening, thus reflecting the intense proteolytic activity of the enzymes contributed by the plant coagulant utilized. The TCASN was also found to be high in this cheese by the end of ripening (16–20%), which suggests a high extent of FAA release throughout maturation. The major FAA by 180 d of ripening were Glu, Val, Leu and Lys, representing 56–70% of the total in all four dairies sampled. Cheeses produced from refrigerated milk possessed higher amounts of γ-amino-n-butyric acid (Gaba) and lower amounts of Glu when compared with those manufactured with non-refrigerated milk.     

Proteolysis and Lipolysis in Ripening Cheddar Cheese Made with Conventional Bulk Starter and with Frozen Concentrated Direct-to-the-Vat Starter Culture

Samples taken at the beginning and after 1, 2, and 3 wk of ripening were assayed for total dipeptidase activity and individual free amino acids. Protein dye-binding values and fat acidity titers were used to assess and monitor proteolysis and lipolysis at 4 days and 1, 2, 3, 6, 9, and 12 months. Results indicated higher dipeptidase activity in the bulk starter cheese for the fist 3 wk of ripening. Total free amino acids at the initial stages of cheese ripening were slightly lower in the cheeses made with the frozen concentrated culture. Studies using the protein-dye binding technique showed significantly higher-dye binding capacity by the 3-month-old cheese made with the bulk starter. The differences, however, became minimal in cheeses at 6, 9, and 12 months of age. Higher fat acidity levels were also obtained in the 3, 6, 9, and 12 month cheeses made with the bulk starter.     

Proteolysis of Mahon cheese as affected by acoustic-assisted brining

 Mahon cheeses were brined in the presence of an ultrasonic field and ripened during 75 days at 12  °C and 85% RH. Secondary proteolysis (water-soluble N, non-protein N, and free amino acids) was measured and compared to that obtained for cheeses conventionally brined. There were no differences in water-soluble and non-protein N attributable to the brining treatment. However, cheeses acoustically brined exhibited higher concentrations of free amino acids. The release of total free amino acids was more pronounced during the first 15 days of ripening for both types of brining treatments. The changes in proteolysis (free amino acids) during cheese ripening caused by acoustic-assisted brining are indicative of a higher extent of proteolysis and may also improve cheese flavor. Received: 13 March 2000     

Influence of native lactic acid bacteria on the microbiological,biochemical and sensory profiles of Serra da Estrela cheese

Cheesemaking from batches of raw ewe's milk was carried out via inoculation with wild strains of Lactococcus lactis subsp. lactis ESB110019 and Lactobacillus plantarum ESB5004 independently, or combined with each other. Those two strains had been isolated from the native microflora of typical Serra da Estrela cheese. One control batch was processed in parallel without addition of any starter. The evolution in viable counts of the main micro-organisms (viz. lactic acid bacteria, Enterobacteriaceae, staphylococci and yeasts), as well as in secondary proteolysis (WSN, 2% TCASN, 12% TCASN and 5% PTASN), was monitored throughout ripening time (over a 63-day period) in cheeses from each batch. The sensory features of the fully ripened cheeses were also assessed. Cheeses manufactured with starter showed significantly lower levels of viable Enterobacteriaceae than those manufactured without starter; viable counts of enterococci and staphylococci did significantly increase after addition of L. lactis or Lb. plantarum, respectively. Proteolysis in terms of WSN and 5% PTASN was not significantly affected by the lactic acid bacteria tested when compared to the control, but L. lactis played a significant role toward increasing the 2% TCASN content of cheeses; both strains led to a statistically significant increase of the 12% TCASN. The scores for flavor and texture of the control cheeses were somewhat above those for the experimental cheeses manufactured with starter.     

Chemometric analysis of proteolysis during ripening of Ragusano cheese

Chemometric modeling of peptide and free amino acid data was used to study proteolysis in Protected Denomination of Origin Ragusano cheese. Twelve cheeses ripened 3 to 7 mo were selected from local farmers and were analyzed in 4 layers: rind, external, middle, and internal. Proteolysis was significantly affected by cheese layer and age. Significant increases in nitrogen soluble in pH 4.6 acetate buffer and 12% trichloroacetic acid were found from rind to core and throughout ripening. Patterns of proteolysis by urea-PAGE showed that rind-to-core and age-related gradients of moisture and salt contents influenced coagulant and plasmin activities, as reflected in varying rates of hydrolysis of the caseins. Analysis of significant intercorrelations among chemical parameters revealed that moisture, more than salt content, had the largest single influence on rates of proteolysis. Lower levels of 70% ethanol-insoluble peptides coupled to higher levels of 70% ethanol-soluble peptides were found by reversed phase-HPLC in the innermost cheese layers and as the cheeses aged. Non-significant increases of individual free amino acids were found with cheese age and layer. Total free amino acids ranged from 14.3 mg/g (6.2% of total protein) at 3 mo to 22.0 mg/g (8.4% of total protein) after 7 mo. Glutamic acid had the largest concentration in all samples at each time and, jointly with lysine and leucine, accounted for 48% of total free amino acids. Principal components analysis and hierarchical cluster analysis of the data from reversed phase-HPLC chromatograms and free amino acids analysis showed that the peptide profiles were more useful in differentiating Ragusano cheese by age and farm origin than the amino acid data. Combining free amino acid and peptide data resulted in the best partial least squares regression model (R(2) = 0.976; Q(2) = 0.952) predicting cheese age, even though the peptide data alone led to a similarly precise prediction (R(2) = 0.961; Q(2) = 0.923). The most important predictors of age were soluble and insoluble peptides with medium hydrophobicity. The combined peptide data set also resulted in a 100% correct classification by partial least squares discriminant analysis of cheeses according to age and farm origin. Hydrophobic peptides were again discriminatory for distinguishing among sample classes in both cases.     

地衣芽孢杆菌凝乳酶对切达干酪成熟过程中蛋白水解的影响

利用地衣芽孢杆菌凝乳酶制作切达干酪和切达干酪类似物,分析干酪成熟过程中各蛋白水解指标的变化规律,以揭示地衣芽孢杆菌凝乳酶对切达干酪成熟过程中蛋白水解的影响。结果表明,CDF组（添加地衣芽孢杆菌D3.11凝乳酶所制切达干酪）、CD3组（添加地衣芽孢杆菌D3.11凝乳酶但未添加发酵剂制成的干酪类似物）和CCF组（添加商品凝乳酶所制切达干酪）干酪蛋白含量、pH 4.6-可溶性氮、12%三氯乙酸-可溶性氮、5%磷钨酸-可溶性氮、总游离氨基酸含量均随着成熟时间延长呈显著增加趋势,并且成熟期间CDF组干酪均显著高于CCF组干酪（P＜0.05）;十二烷基硫酸钠-聚丙烯酰氨凝胶电泳分析表明,CDF组干酪α-酪蛋白水解程度较大;pH 4.6-可溶性肽段分析表明,随着干酪的成熟,总肽含量呈先增加后下降趋势,但疏水性肽与亲水性肽的比值呈持续下降趋势,在成熟第6个月时,CDF组、CD3组和CCF组干酪疏水性肽与亲水性肽比值分别为2.668、2.822、3.788。主成分分析表明,3 组干酪的蛋白水解程度与成熟度呈正相关,与疏水性肽和亲水性肽的比值呈负相关。以上结果表明,利用地衣芽孢杆菌凝乳酶制作的干酪蛋白水解度更高,但其疏水性肽比例较小,研究结果可为地衣芽孢杆菌凝乳酶在干酪生产中的应用提供理论依据。