基于多元线性回归对Tilsit型干酪成熟期的研究

根据Tilsit干酪加工方法制作生干酪样品,在90 d成熟内测定其总氮、水溶性氮(WSN)、三氯乙酸水溶性氮(TCA-SN)、磷钨酸水溶性氮(PTA-SN)等含量,研究成熟过程中蛋白质水解程度。上述指标,除磷钨酸水溶性氮,其余均与干酪成熟期呈现一定的线性相关性(p<0.05),该多元线性回归模型可以用于鉴定和预测Tilsit型干酪的成熟期。     

Effects of somatic cell count and stage of lactation on the plasmin activity and cheese-making properties of ewe milk

The experiment was conducted from March to July 2002 using 5 intensively managed flocks of Southern Italy. In each flock, 2 groups of 50 ewes were created. The groups were designated LSCC (low somatic cell count [SCC]) when their milk SCC was lower than 500,000/mL and HSCC (high SCC) when their milk SCC was higher than 1,000,000/mL. Bulk milk and whey samples were analyzed for fat, total protein, lactose, casein, and whey protein contents. Renneting properties of milk were also determined. Moisture, NaCl, and nitrogen fractions were determined in fresh cheese curds. In addition, plasmin (PL) and plasminogen (PG) activities in milk and cheese were monitored. The proteolytic activity of plasmin by urea-polyacrylamide gel electrophoresis and the white blood cell (WBC) differentials were determined. The HSCC resulted in higher pH values in milk and in higher moisture and lower fat contents in fresh cheese curds. Moreover, a lower recovery of fat and whey proteins was obtained from the HSCC than from the LSCC raw milk. The crude protein and casein contents were higher in the HSCC than in the LSCC curds during early and midlactation; an opposite trend was observed in late lactation. Plasmin and PG activities underwent more marked fluctuations in the LSCC than in the HSCC curds through lactation. The results of this experiment demonstrate that the PL activity in ewe milk is markedly influenced by the SCC, although SCC is not the only parameter for predicting PL and PG evolution in ewe milk. The LSCC milk resulted in a higher proteolytic potential of Canestrato pugliese cheese curds.     

Comprehensive analysis of proteolysis during 8 months of ripening of high-cooked Old Saare cheese

We applied capillary electrophoresis, liquid chromatography coupled with tandem mass-spectrometry (MS/MS), and ultra-performance liquid chromatography to determine the composition of water-insoluble and water-soluble proteinaceous fractions of the cheese and to study in detail the degradation of caseins during 8 mo of ripening of Estonian high-temperature cooked hard cheese Old Saare. The application of high-resolution and high-accuracy MS/MS enabled identification of more than 3,000 small peptides, representing a fairly full casein peptidome containing peptides of 4 to 25 AA in length: 1,049 from β-casein (CN), 944 from α_S1-CN, 813 from α_S2-CN, and 234 from κ-CN. The majority of β-CN- and α_S1-CN-derived peptides originated from the N-terminal parts of the molecule, f6-93 and f1-124, respectively; peptides from α_S2-CN arose predominantly from the C-terminal end f100-162. At the beginning of ripening, we found a relatively high amount of peptides originating from the glycomacropeptide part of κ-CN, whereas peptides from para-κ-CN prevailed during the later stages of ripening of the cheese. The cleavage patterns of β-CN, α_S2-CN, as well as α_S1-CN, showed that primary proteolysis was started mainly by plasmin, although a low proteolytic activity of chymosin was also evident. Based on the analysis of cleavage sites, we observed a significant participation of proteolytic enzymes, including amino- and carboxypeptidases, of both mesophilic and thermophilic starter bacteria in further hydrolysis of oligopeptides during the ripening. Several new phosphopeptides were detected in the result of MS/MS data analysis. The profiles of the estimated concentrations of phosphopeptides revealed that those originating from β-CN and α_S1-CN accumulated during cheese maturation. In contrast, we did not notice any generation of phosphopeptides from the highly phosphorylated part of α_S2-CN, f25-80, presumably due to the inaccessibility of this region to the action of plasmin and chymosin. The analysis of cleavage sites and the combination of principal component and clustering analyses provided a characterization of the complex dynamics of formation and degradation of peptides during cheese maturation. We made an attempt to obtain a comprehensive picture of proteolysis during Old Saare cheese ripening on the basis of the detailed peptidomic data, including also the less abundant peptides determined by MS/MS, and complemented by the data on intact caseins and free AA and reported the results in the paper.     

提高成熟温度加快Mozzarella干酪成熟的研究

制作2批Mozzarella干酪A和B，分别在4℃(A)和7℃(B)下成熟，观察其在成熟期间的变化及测定可溶性N的含量等指标，可知在7℃下成熟的干酪在制作后30d的蛋白水解性、功能特性等和4℃下成熟50d的干酪无显著差异，而和7℃下成熟50d的干酪有显著差异。说明成熟温度显著影响干酪的蛋白水解性。在7℃下贮藏的Mozzarella干酪成熟30d可达到4℃下贮存50d的成熟度，即将成熟温度从4℃提高到7℃，可将成熟期缩短20d左右。     

凝乳酶对牦牛乳硬质干酪成熟期间蛋白降解的影响

以新鲜牦牛乳为原料,采用小牛皱胃酶、木瓜蛋白酶和微生物凝乳酶制作硬质干酪,探讨凝乳酶种类对牦牛乳硬质干酪成熟期间蛋白质降解的影响。结果表明:三种凝乳酶牦牛乳硬质干酪成熟过程中,不同凝乳酶牦牛乳硬质干酪在成熟期间蛋白质降解能力存在较大差异,总氮（TN）、p H4.6水溶性氮（p H4.6-SN/TN）、12%的三氯乙酸氮（12%TCA-N/TN）、5%磷钨酸氮（5%PTA-N/TN）含量、游离氨基酸均随成熟时间延长不同程度的增加,蛋白氮和酪蛋白氮逐渐降低,多肽氮呈先升高后下降趋势,且微生物凝乳酶降解牦牛乳硬质干酪蛋白能力显著（p<0.05）高于木瓜蛋白酶和小牛皱胃酶。      

快速成熟契达干酪成熟期间的理化特性变化

对天然契达(Cheddar)干酪快速成熟期间理化特性进行了研究,结果表明,在21 d内,pH值呈下降趋势(P<0.05),在21 d到35 d的成熟中,pH值缓慢增加(P>0.05);Cheddar干酪快速成熟过程中由于蛋白酶和脂肪酶的作用使蛋白发生水解,pH值为4.6醋酸溶液—水溶性氮质量分数、质量分数为12%的三氯乙酸—可溶性氮和游离氨基酸(FAA)的质量分数都呈明显增加趋势(P<0.05);成熟过程中TPA各项指标均有不同程度的变化,硬度和凝聚性呈先下降后增加的趋势,弹性和咀嚼性呈先上升后下降的趋势。     

成熟温度对Mozzarella干酪蛋白水解和质构的影响

研究了温度为4,7,10℃时对干酪成熟过程中蛋白水解和质地的影响。结果表明,随着干酪成熟温度的升高,成熟期间干酪中可溶性氮与总氮的比值增加较快,干酪的硬度下降速度也较快。说明在较高的成熟温度下,干酪在较短的时间内能够达到成熟的状态。     

Volatile composition and proteolysis in traditionally produced mature Kashar cheese

Twelve samples of raw milk mature Kashar cheese at different stages of ripening were collected from retail outlets. The average pH, moisture, fat-in-dry matter, protein, salt-in-dry matter and titratable acidity contents of the samples were 5.33, 39.39%, 45.20%, 27.33%, 6.62% and 0.65% (as lactic acid), respectively. Indices of proteolysis varied from 10.72% to 23.75% and 7.09% to 12.26% for pH 4.6-soluble and 12% trichloroacetic acid-soluble nitrogen fractions, respectively, and total free amino acid concentrations ranged from 6.36 to 36.03 mg Leu g⁻¹ of cheese. The cheeses were analysed for volatile compounds by Solid Phase Microextraction and Gas Chromatography-Mass Spectrometry (GC-MS). A total of 113 compounds were detected and identified belonging to the following chemical groups: acids (eleven), esters (sixteen), ketones (sixteen), aldehydes (six), alcohols (twenty-seven), sulphur compounds (seven), terpenes (seven) and miscellaneous compounds (twenty-three). The potential effect of each compound on the flavour profile of Kashar cheese is discussed. Acids, esters, ketones and alcohols were found at considerable levels in the samples. Kashar cheeses obtained from different retail outlets displayed some differences in terms of chemical composition, proteolysis and patterns of aroma compounds; and may be attributed to their production technologies and age-related variations.     

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.     

Chymosin-mediated proteolysis, calcium solubilization, and texture development during the ripening of cheddar cheese

Full fat, milled-curd Cheddar cheeses (2 kg) were manufactured with 0.0 (control), 0.1, 1.0, or 10.0 μmol of pepstatin (a potent competitive inhibitor of chymosin) added per liter of curds/whey mixture at the start of cooking to obtain residual chymosin levels that were 100, 89, 55, and 16% of the activity in the control cheese, respectively. The cheeses were ripened at 8°C for 180 d. There were no significant differences in the pH values of the cheeses; however, the moisture content of the cheeses decreased with increasing level of pepstatin addition. The levels of pH 4.6-soluble nitrogen in the 3 cheeses with added pepstatin were significantly lower than that of the control cheese at 1 d and throughout ripening. Densitometric analysis of urea-PAGE electro-phoretograms of the pH 4.6-insoluble fractions of the cheese made with 10.0 μmol/L of pepstatin showed complete inhibition of hydrolysis of α_S1-casein (CN) at Phe₂₃-Phe₂₄ at all stages of ripening. The level of insoluble calcium in each of 4 cheeses decreased significantly during the first 21 d of ripening, irrespective of the level of pepstatin addition. Concurrently, there was a significant reduction in hardness in each of the 4 cheeses during the first 21 d of ripening. The softening of texture was more highly correlated with the level of insoluble calcium than with the level of intact α_S1-CN in each of the 4 cheeses early in ripening. It is concluded that hydrolysis of α_S1-CN at Phe₂₃-Phe₂₄ is not a prerequisite for softening of Cheddar cheese during the early stages of ripening. We propose that this softening of texture is principally due to the partial solubilization of colloidal calcium phosphate associated with the para-CN matrix of the curd.     

不同包装方式对半硬质干酪成熟期间蛋白质降解的影响

对真空包装和涂蜡包装的半硬质干酪成熟过程中蛋白降解进行了研究。结果表明:2种包装的干酪在成熟过程中pH 4.6SN含量和12%TCA-SN含量都随着时间的延长逐渐增大,且2组数据之间差异显著(P<0.05);2种包装的干酪中游离氨基酸总量随成熟时间的延长而逐渐增加,各种氨基酸含量变化的显著性不同;SDS-PAGE电泳图谱显示2种干酪在成熟期内蛋白质都发生了明显的降解,且涂蜡包装的干酪蛋白降解程度较真空包装的深,在成熟45 d后较为明显。     

Influence of calcium and phosphorus, lactose, and salt-to-moisture ratio on Cheddar cheese quality: proteolysis during ripening

Proteolysis in cheese is influenced by the state of proteins (protein-calcium-phosphate interactions), level of indigenous milk enzymes (plasmin), externally added milk-clotting enzymes (chymosin), and endogenous and exogenous enzymes from starter and non-starter lactic acid bacteria (NSLAB). The objective of this study was to determine how different levels of calcium (Ca) and phosphorus (P), residual lactose, and salt-to-moisture ratio (S/M) in cheese influence proteolysis during ripening. Eight cheeses with 2 levels of Ca and P (0.67 and 0.47% vs. 0.53 and 0.39%, respectively), 2 levels of lactose at pressing (2.4 vs. 0.78%), and 2 levels of S/M (6.4 vs. 4.8%) were manufactured. The cheeses were analyzed for changes in pH 4.6-soluble N, and starter and NSLAB counts during 48 wk of ripening. Cheeses at d 1 were also analyzed for residual chymosin, plasmin, and plasminogen activity. A significant increase in soluble N was observed during ripening for all the treatments. Cheeses with low Ca and P, low lactose, and low S/M treatments exhibited higher levels of proteolysis as compared to their corresponding high treatments. Differences in the rate of proteolysis for cheeses with different levels of Ca and P might be due to changes in protein conformation and differences in residual chymosin in the cheeses. Cheeses with low Ca and P were manufactured by lowering the pH at set and drain, which led to higher chymosin retention in cheeses with low Ca and P compared with high Ca and P. Differences in proteolysis between treatments with different levels of lactose were also partly attributed to residual chymosin activity. In all treatments, a major fraction of plasmin existed as plasminogen, indicating minimal contribution of plasmin to proteolysis in Cheddar cheeses. The number of starter bacteria, in all treatments, decreased significantly during ripening. However, the decrease was larger in the case of high S/M treatments compared with low S/M treatments. In contrast, the number of NSLAB increased during ripening, and low S/M cheeses had higher counts compared with high S/M cheeses. The differences in proteolysis due to S/M were partially attributed to changes in protein conformation or bacterial proteolytic activity.     

非发酵剂乳酸菌A-3和D-3对半硬质山羊奶干酪成熟的影响

目的获得加速半硬质山羊奶干酪成熟的非发酵剂乳酸菌菌株(non-starterlacticacidbacteria,NSLAB)。方法以前期分离自地中海地区山羊奶干酪中的2株优良NSLAB菌株为研究对象,测定其对干酪成熟过程中组成成分、微生物菌群、蛋白质水解和质构的影响。结果添加NSLAB菌株对干酪组成成分没有显著影响, NSLAB菌株没有影响乳球菌生长,在干酪成熟期间pH 4.6-SN和12%TCA-SN逐渐增加,且添加NSLAB的干酪在成熟30 d后显著增加了pH 4.6-SN和12%TCA-SN含量, 5%PTASN/TN的增加主要是由于乳酸菌中肽酶作用的结果, SDS-PAGE电泳结果说明添加NSLAB菌株的干酪中小分子多肽含量明显比对照干酪多,RP-HPLC分析得出干酪水溶性中肽的数量随着成熟时间增加。添加NSLAB菌株A-3没有改变干酪的硬度,使干酪的弹性增加。结论添加菌株A-3作为NSLAB的干酪样品中微生物自溶率高,蛋白水解程度强,质构性能良好,具有加速干酪成熟的潜力,是山羊奶干酪工业化生产的优良NSLAB。     

Proteolytic and lipolytic changes during the ripening of a Spanish craft goat cheese (Armada variety)

The proteolytic and lipolytic changes during the ripening process were investigated in four batches of Armada goat's milk cheese (an artisanal variety produced in the North of Spain), by determining the classical nitrogen fractions, caseins and their degradation products, free amino acids, as well as the acidity of the fat, thiobarbituric acid (TBA) number and free fatty acids. Values obtained for the nitrogen fractions and for caseins and their degradation products show that this cheese undergoes very little protein degradation. A low free amino acids content was observed throughout the ripening process with a predominance of Pro followed by Leu+Ile, Glu acid, Phe, His+Lys and Val. The lipid degrada-tion was very intense from the second month of ripening, only comparable to that reported for cheeses ripened by moulds. The average free fatty acids content increased 20-fold during ripening, reaching final values of 44·5 g kg⁻¹. All the free fatty acids increased considerably during ripening, resulting in a predominance of saturated and unsaturated long-chain acids, followed by medium-chain acids, C₁₀ principally. Short-chain fatty acid content by the end of ripening was higher than that presented in other cheese varieties with a similar high degree of lipolysis. © 1997 SCI.     

Effect of proteolysis and calcium equilibrium on functional properties of natural cheddar cheese during ripening and the resultant processed cheese

The changes in proteolysis, calcium (Ca) equilibrium, and functional properties of natural Cheddar cheeses during ripening and the resultant processed cheeses were investigated. For natural Cheddar cheeses, the majority of the changes in pH 4.6 soluble nitrogen as a percentage of total nitrogen (pH 4.6 SN/TN) and the soluble Ca content occurred in the first 90 d of ripening, and subsequently, the changes were slight. During ripening, functional properties of natural Cheddar cheeses changed, that is, hardness decreased, meltability was improved, storage modulus at 70 °C (G'T=70) decreased, and the maximum tan delta (TDmax) increased. Both pH 4.6 SN/TN and the soluble Ca were correlated with changes in functional properties of natural Cheddar cheeses during ripening. Kendall's partial correlation analysis indicated that pH 4.6 SN/TN was more significantly correlated with changes in hardness and TDmax. For processed cheeses manufactured from natural Cheddar cheeses with different ripening times, the soluble Ca content did not show significant difference, and the trends of changes in hardness, meltability, G'T=70, and TDmax were similar to those of natural Cheddar cheeses. Kendall's partial correlation analysis suggested that only pH 4.6 SN/TN was significantly correlated with the changes in functional properties of processed cheeses.     

Effect of parity and stage of lactation on feed sorting behavior of lactating dairy cows

The objectives of this study were to determine if feed sorting differs between primiparous (PP) and multiparous (MP) cows, if sorting changes from the period of peak lactation to peak dry matter intake (DMI), and whether feed sorting affects efficiency of production. Data on DMI, milk production, feed sorting (particle size of offered and refused feed), and energy status (plasma nonesterified fatty acid, β-hydroxybutyrate, insulin and glucose concentration) were collected on 30 PP and 30 MP lactating Holstein dairy cows, individually housed and fed in tie-stalls, during 3 wk (wk 2, 6, and 10) over 10 wk of a lactation study. Cows averaged 53, 81, and 109 ± 10 d in milk (DIM) at the beginning of each of the 3 recording weeks. To determine sorting, feed samples were separated with a particle separator that had 3 screens (19, 8, and 1.18 mm) and a bottom pan, resulting in 4 fractions (long, medium, short, fine). Over the study period, MP cows consumed more DM and produced more milk than did the PP cows, but had similar efficiency of production (kg of milk/kg of DMI), and similar levels of plasma indicators of energy status. Across the study period, DMI increased, whereas milk yield decreased, resulting in decreased efficiency of milk production as cows moved further into lactation. All cows had higher nonesterified fatty acid and lower insulin concentrations in plasma at 53 DIM compared with at 81 and 109 DIM, suggesting they were mobilizing more body fat at that earlier stage of lactation. Across periods, all cows sorted against the longest ration particles, did not sort the medium ration particles, and sorted for fine ration particles; as a result, all cows consumed less neutral detergent fiber (NDF) and physically effective NDF (proportion of NDF retained on the 19- and 8-mm screens of the particle separator) than predicted. Greater selection against the longest ration particles was associated with greater efficiency of milk production; however, this sorting pattern also tended to be associated with lower milk fat percentage. The extent of the observed sorting against the longest, physically effective ration particles and for the finest ration particles was greater for PP cows than for MP cows across all 3 observation periods. Feed sorting remained consistent in cows across the DIM evaluated in this study and that this sorting behavior can affect the efficiency of milk production as well as milk fat percentage. Further, PP cows engage in more sorting of their ration than MP cows.     

干酪成熟过程中发酵剂的作用

介绍干酪成熟过程中乳酸菌发酵剂及二级发酵剂的作用,包括在干酪风味、质地、加速干酪成熟、产生抗菌素及营养和颜色等多方面作用,同时分析成熟期间发酵剂控制不当会引起的负面作用,提出应用具有选择性的发酵剂或经遗传修饰的发酵剂生产干酪具有较强的应用前景。     

Composition,proteolysis, and volatile profile of Strachitunt cheese

Strachitunt, a blue-veined Italian cheese, received the Protected Designation of Origin (PDO) label in 2014. Its unique technological feature is represented by the dual-curd method of production. Strachitunt is produced from raw bovine milk with or without the inoculation of natural starter cultures of lactic acid bacteria, and the addition of secondary cultures of mold spores is not permitted by the product specification. Physico-chemical properties, proteolysis, and volatile profile of Strachitunt were investigated in 10 cheese samples (ripened for 75 d) made throughout spring 2015 and provided by the main cheese maker. Overall, composition parameters showed a large variability among samples. Cheese was characterized by an acid paste (pH 5.46) and a lower extent of proteolysis compared with other blue-veined varieties. The main chemical groups of volatile organic compounds were alcohols and esters, whereas ketones represented only a minor component. The erratic adventitious contamination by mold spores of the cheese milk, the unique dual-curd method of cheese-making, and the large time variability between the piercing time and the end of ripening could be highlighted as the main causes of both the distinctive analytical fingerprint and the scarce standardization of this blue-veined cheese.