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。     

Effect of partial substitution of NaCl with KCl on proteolysis of halloumi cheese

The effect of substitution of NaCl with Potassium chloride (KCl) in brine solution on proteolysis of halloumi cheese was investigated. Halloumi cheeses were made and kept in 4 different brine solutions (18% w/w), including only NaCl (HA; control); 3NaCl:1KCl (w/w) (HB); 1NaCl:1KCl (w/w) (HC); 1NaCl:3KCl (w/w) (HD); and stored for 56 d at 4 °C. Proteolysis was assessed using water-soluble nitrogen (WSN), trichloroacetic acid-soluble nitrogen (TCA-SN), phosphotungstic-soluble nitrogen (PTA-SN), urea polyacrylamide gel electrophoresis (urea-PAGE), and peptide patterns. WSN and TCA-SN contents were similar in all experimental cheeses. Peptide patterns of the pH 4.6 N fraction and urea-PAGE showed no significant difference between halloumi cheeses kept in various NaCl/KCl mixtures (HB, HC, HD) and control (HA). Sodium and potassium contents showed positive correlations with WSN and PTA-SN. There was an inverse correlation between calcium (Ca) contents and WSN and PTA-SN. Correlations between Ca and Na or K were negative at the same salt treatment.     

Effect of freeze–dried kefir culture on proteolysis in feta-type and whey-cheeses

The effect of freeze–dried kefir culture on the proteolysis of feta-type and whey-cheese was investigated. All nitrogen fractions increased during ripening. Although no significant differences were observed in total nitrogen (TN), the levels of water-soluble nitrogen (WSN), pH 4.4-soluble nitrogen (SN), 12% trichloroacetic acid-soluble nitrogen (TCA-SN) and phosphotungstic acid-soluble nitrogen (PTA-SN) were significantly higher in cheeses produced by freeze–dried kefir culture during the later stages of ripening. Content of total free amino acids (FAA) was significantly affected by freeze–dried kefir starter culture and it was continuously increased in kefir-cheese while, in rennet-cheese it was increased up to 30 days of ripening and then slightly decreased. On the other hand, FAA content continuously decreased in kefir-whey-cheese whereas it increased in whey-cheese. The cheese samples produced by freeze–dried kefir as starter culture were characterised as high-quality products during the preliminary sensory evaluation and they were accepted by the panel. Overall, the use of freeze–dried kefir suggested acceleration of cheese ripening and resulted in improved sensory characteristics.     

干酪介电特性与成熟期及成熟度相关性分析

以半硬质契达干酪为研究对象,对同一加工样品在9℃贮存成熟,分别对成熟期0、15、30、45、60、90、120、150、180 d的干酪样品进行了介电特性和成熟度指标的测定,包括干酪样品总氮(total nitrogen,TN)含量、p H 4.6可溶性氮(soluble nitrogen,SN)含量及三氯乙酸(trichloroacetic acid,TCA)溶液中SN含量的测定。测试频率选定为500、915、1 500、2 000 MHz,对测试结果进行了统计分析,建立了干酪介电特性与成熟度指标之间的相关性。结果表明:随着成熟期的延长,干酪水分含量略有减少、水分活度明显降低、p H值先降低后升高、成熟度指数p H 4.6-SN/TN和12%TCA-SN/TN均随成熟期延长而增大;介电常数ε’在所选的测试频率下与成熟期和成熟度指数的变化均呈线性相关关系;介电损耗因数ε’’在所选测试频率范围内,随成熟期的延长和成熟度的增大呈现总体下降趋势,与成熟期和成熟度在500 MHz和915 MHz频率条件下线性相关性极显著(P0.01),而在高频1 500 MHz和2 000 MHz条件下线性相关性不显著;损耗角正切值变化不明显,表明在测试频率范围介电常数ε’和介电损耗因数ε’’的变化方向一致,同时变化幅度相近。     

高压处理的霉菌干酪的蛋白水解

对自制的白霉干酪采用高压处理工艺,分析其蛋白水解的情况,并与未经高压处理干酪进行比较。通过测定2种干酪表面和内部的pH4.6-SN(%TN)、12%TCA-SN(%TN)和FAA(mg/g干酪)评价蛋白水解程度,结果表明:高压处理使pH4.6-SN、12%TCA-SN增加,对FAA无影响;高压处理使pH值升高,影响部分微生物酶的活力;高压处理能降低疏水作用,使酶更容易作用于暴露的敏感键。     

一种毛霉表面成熟干酪的蛋白质水解作用评价

从毛豆腐中分离出一株毛霉,并应用于表面成熟干酪,以研究干酪成熟过程中所发生的蛋白质水解作用。在90d 的成熟过程中,干酪的pH 值增加;蛋白质水解作用的评价指标,如干酪外层的水溶性氮- 总氮比、pH4.6水溶性氮- 总氮比、12g/100mL 三氯乙酸可溶性氮- 总氮比,在成熟90d 后分别增加至(23.68 ± 1.07)%、(19.38 ± 1.32)%和(8.61 ± 0.85)%,并且高于干酪的内部相应指标。SDS-PAGE 和毛细管电泳分析干酪的pH4.6 不溶性组分,结果表明酪蛋白在干酪成熟过程中被降解。对干酪成熟过程中分离出的水溶性组分进行RP-HPLC 分析,结果显示成熟过程中蛋白质被水解以及形成一些新肽分子。     

Ripening of traditional Örgü cheese manufactured with raw or pasteurized milk: Composition and biochemical properties

The changes in composition and some biochemical properties of Örgü cheeses made from raw (RMC) and pasteurized (PMC) cow milk were investigated during a 90-day ripening period. The average contents of total solids (TS), protein, water soluble nitrogen (WSN), trichloro-acetic acid soluble nitrogen (TCA-SN) and acid degree value (ADV) were lower, while salt and salt in TS were found to be statistically higher in PMC than RMC (P < 0.05). In addition, in both RMC and PMC, the TS and protein contents were decreased as compared to an increase in salt, salt in TS, WSN and TCA-SN contents, and ADV, during ripening (P < 0.05). The evaluation of WSN, TCA-SN and ADV shows that these two experimental Örgü cheese types undergo little proteolysis and lipolysis. On the other hand, acidity development was observed to be high in both before curdling and in cheese made from raw milk during ripening.     

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。     

溶菌酶处理发酵剂加速干酪成熟的研究

以pH、蛋白质降解、脂肪酸含量变化及干酪的感官特征为指标,研究了溶菌酶处理发酵剂对干酪成熟的影响。结果表明:实验组干酪的pH在整个成熟期处于对照组1(添加2%发酵剂)干酪和对照组2(添加2.5%发酵剂)的pH之间,表明通过溶菌酶处理发酵剂可明显抑制干酪成熟期间的产酸量;添加溶菌酶处理发酵剂可加速干酪中蛋白质分解和乳脂肪的降解;感官评价结果表明,添加溶菌酶处理发酵剂可提高干酪风味,在相同的成熟时间实验组干酪的质地也较对照组好。     

Proteolysis in miniature Cheddar-type cheeses made using blends of chymosin and Cynara cardunculus proteinases as coagulant

Miniature (20 g) Cheddar-type cheeses were manufactured using blends of Cynara cardunculus proteinases and chymosin as coagulant (100 : 0, 50 : 50, 25 : 75 and 0 : 100 C. cardunculus proteinases : chymosin). There were no substantial differences between the compositions of cheeses made using any of the four coagulant blends. Cheeses manufactured with coagulant blends containing C. cardunculus proteinases exhibited higher levels of pH 4.6-soluble nitrogen than cheese made using chymosin as coagulant. The extent of breakdown of α _s1 -casein, as measured by urea-polyacrylamide gel electrophoresis (urea-PAGE), was greater in cheeses made using coagulant preparations containing C. cardunculus proteinases as a constituent than in cheese made using 100% chymosin as coagulant. Different reverse-phase high-performance liquid chromatography (RP-HPLC) peptide profiles of the ethanol-soluble and -insoluble fractions were obtained for cheeses made using either C. cardunculus proteinases or chymosin as coagulant. Principal component analysis and hierarchical cluster analysis of RP-HPLC data confirmed that the inclusion of even small proportions (25%) of C. cardunculus proteinases with chymosin in the coagulant blend greatly altered the pattern and extent of proteolysis in miniature Cheddar-type cheeses.     

开菲尔直投式发酵剂在益生菌干酪生产中的应用研究

该研究主要将开菲尔粒制备成直投式发酵剂应用于干酪的生产。 以FD-DVS R-704商业干酪发酵剂制作的干酪为对照,对其 理化指标、挥发性香气成分、质构特性和感官指标进行测定与评价。 结果表明,与对照干酪相比,开菲尔干酪的水分含量、pH4.6-可溶 性氮（SN）及12%三氯乙酸（TCA）-SN的含量极显著增高（P＜0.01）,pH值极显著低（P＜0.01）,而粗脂肪比例和总游离氨基酸（TFAA） 含量无显著差异（P＞0.05）,不饱和脂肪酸（UFA）含量显著增高（P＜0.05）;醇类和酯类物质的种类增多;硬度和咀嚼性极显著降低（P＜0.01）,胶黏性极显著增大（P＜0.01）,内聚性和弹性无明显差异（P＞0.05）;开菲尔粒益生菌干酪呈乳白色,表面有光泽,不仅具 有干酪特有的滋味和气味,且具有开菲尔发酵乳的特有风味,带有适宜的酒香味。     

A Survey on the Some Chemical and Biochemical Properties of Civil Cheese,a Traditional Turkish Cheese

In this article, 15 randomly selected samples of Civil cheese, were purchased from different retail markets in the Erzurum province, Turkey and were investigated for some chemical and biochemical analyses. All cheese samples were analyzed for dry matter, fat, salt, ash, titrable acidity, total nitrogen, soluble nitrogen, ripening index, α_s-and β-casein degradation, γ-casein, and peptides. Dry matter, fat, fat in dry matter, salt, salt in dry matter, ash, and acidity values in samples analyzed were found to be as found between 31.33 and 40.12 g/100 g cheese; 1.00 and 7.00 g/100 g cheese; 2.49 and 18.98 g/100 g cheese; 0.11 and 0.34 g/100 g cheese; 0.27 and 1.04 g/100 g cheese; 1.42 and 5.14 g/100 g cheese and, 0.63 and 2.16%, respectively. TN, WSN/TN, TCA-SN/TN, and PTA-SN/ TN values, expressed as TN%, were found between 3.01 and 5.57 g/100 g cheese, 4.25 and 8.80 g/100 g cheese, 3.23 and 6.12 g/100 g cheese, 1.03, and 5.53 g/100 g cheese in Civil cheese samples analyzed, respectively. SDS-PAGE showed that both α_s-CN and β-CN ratios were not high compared with similar cheeses, and are not completely hydrolyzed in all Civil cheese samples. A broad range of values from chemical and biochemical analysis indicated that Civil cheeses collected from retail markets lacked standardization. Consequently, it was decided that Civil cheese samples do not undergo an excessive proteolysis.     

A Survey on Selected Chemical,Biochemical and Sensory Properties of Kes Cheese,A Traditional Turkish Cheese

Kes, a traditional Turkish dairy product, is commonly produced in the northern Anatolian region of Turkey. Kes samples were obtained from local markets in Ordu vicinity and were investigated for some chemical, biochemical, and sensorial analyses. The mean values of dry matter (DM), fat, fat in DM, salt, salt in DM, ash, and pH were found to be 56.17 ± 6.07, 8.79 ± 2.84, 15.78 ± 5.39, 3.22 ± 1.35, 5.68 ± 2.19, 4.31 ± 1.29 g/100 g cheese, and 4.75 ± 0.59, respectively. The WSN/TN, TCA-SN/TN and PTA-SN/TN values were between 1.79–28.53, 1.67–23.24, and 0.44–17.56%, respectively. SDS-PAGE showed that both α_s–CN and β–CN fractions were not completely hydrolyzed in all Kes cheese samples. α_s–CN, β–CN, and γ-casein and other peptides fractions determined as 34.57 ± 5.21, 33.74 ± 4.21, and 31.37 ± 6.82%, respectively. Panelists gave high scores (above 7 out of 10) to all samples for all the attributes considered.     

热烫拉伸温度对Mozzarella干酪蛋白降解及质构特性的影响

不同的热烫拉伸温度对Mozzarella干酪的组织结构、凝块中微生物以及酶的活性有很重要的影响,进而会影响其成熟过程中蛋白质的降解以及最终品质的形成,该研究对不同热烫拉伸温度的Mozzarella干酪成熟过程中的蛋白降解及质构特性指标进行测定。结果表明,同一成熟期内,随着热烫温度（对照、70 ℃、90 ℃）的提高,干酪的pH 4.6可溶性氮（SN）显著下降（P＜0.05）,12%三氯乙酸-可溶性氮（TCA-SN）有所下降（P＞0.05）,而干酪的硬度、咀嚼性均显著增大（P＜0.05）,弹性增大而黏附性有所下降（P＞0.05）,融化性、油脂析出性均显著下降（P＜0.05）。因而采取70 ℃的热烫拉伸温度更有利于干酪成熟后形成良好的质构特性及充分的蛋白降解。     

Production of hard cheese from caprine milk by the use of two types of probiotic cultures as adjuncts

Hard cheeses (Kefalotyri-like) were manufactured from caprine milk with yoghurt as a starter (A), and with its partial replacement with the probiotic adjuncts Lactobacillus rhamnosus LC 705 (B) and/or Lactobacillus paracasei ssp. paracasei DC 412 (C). Both adjuncts retarded the growth of enterococci, and the environment in cheese B did not favour the recovery of lactic acid bacteria (LAB) on Rogosa agar. However, better recovery of the LAB population on M17 agar from cheeses B and C made with adjuncts was recorded early in ripening, and this was accompanied by a greater decrease in pH. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) of whole-cell protein demonstrated that cheese C, made with Lb. paracasei ssp. paracasei as adjunct, is a better vehicle for delivery of live probiotic cells (10 ⁷   cfu/g) to the gastrointestinal tract than cheese B, made with Lb. rhamnosus ; the latter did not belong to the predominant microflora of one out of the two B cheeses. Urea-PAGE electrophoresis results indicated that adjunct lactobacilli enhanced the degradation of both α _S -casein (α _S -CN) and β-casein (β-CN). In the fresh cheese, hydrolysis of α _S -CN was more rapid than β-CN, and the free amino acid content of B and C was higher than in A. Lipolysis products were also higher in B and C than in A as ripening progressed, and the organoleptic characteristics of these cheeses resulted in higher scores, in the order C > B > A. Thus, making Kefalotyri-like cheese from caprine milk with probiotic lactobacilli, particularly Lb. paracasei ssp. paracasei, as adjunct can be considered an effective way of producing a cheese with a large number of probiotic cells.     

Minimization of aflatoxin M1 content in Iranian white brine cheese

A chemometric approach was used to minimize the aflatoxin M1 (AFM1) content of Iranian white brine cheese. The effects of processing factors such as renneting temperature (30–40°C), cutting size (0.5–1 cm), stirring time (10–20 min), press time (1–2 h), curd size (64–256 cm³) and saturated brine pH (4.6–6) on the AFM1 content of the cheese curds were explored. Renneting temperature, press time and saturated brine pH were, respectively, the most significant factors. The aflatoxin content of the cheese samples decreased with increasing renneting temperature and press time. Lowering of the saturated brine pH reduced AFM1 in the cheese curds. Taking account of all of the factors studied, optimum processing conditions for minimization of AFM1 in the cheese curds were: renneting temperature = 39.91°C, cut size = 0.51 cm, stirring time = 17.71 min, press time = 19.48 min, curd size = 73.27 cm³ and saturated brine pH = 4.79.     

Ripening of Camembert-type cheese made from caprine milk using calf rennet or kid rennet as coagulant

Camembert-type cheese was made from caprine milk using either calf rennet or kid 'Grandine' rennet as coagulant. The pH of all cheeses increased throughout ripening and levels of pH 4.6-soluble nitrogen increased from 8.1 to 18.2% of total nitrogen (TN) and from 6.9 to 20% TN for the cheeses made using calf rennet and kid rennet, respectively. Degradation of β-casein, measured by urea–polyacrylamide gel electrophoresis, and total and free amino acids were greater in the cheese made using kid rennet. Production of peptides, analysed by high performance liquid chromatography (HPLC), was slightly more extensive in the Camembert-type cheese made using calf rennet as coagulant. In general, a higher degree of proteolysis was found in Camembert-type cheese made from caprine milk using kid rennet than in cheese made using calf rennet as coagulant.     

Rennin-like milk coagulant enzyme produced by a local isolate of Mucor

Among 20 isolates of Mucor isolated from various environments in Jordan and found to produce a rennin-like acid protease, known as Mucor rennin-like enzyme (MRE), Mucor J20 was found to produce the highest level of MRE. The optimum incubation conditions for enzyme production in a fortified wheat bran mixture using solid-state fermentation were 3–4 days at 30°C. The highest MRE activity (185–200 rennin units or RU) was produced in a medium containing wheat bran and lentil straw (1 : 1 w/w) moistened with whey, and incubated in clay pots at 30°C for 4 days. A slightly lower activity value (178 RU) was found when using a mineral salt solution or distilled water instead of whey, or when using wheat bran alone with whey. At pH 4, the MRE retained its complete activity (100%) for 6 weeks at 5°C and 10°C, and for 3 and 2 weeks at 20°C and 30°C, respectively. After heating at 60°C for 10 min, the enzyme lost its activity at all pH levels used (pH 2–8). The crude extract of MRE was successfully applied in the manufacture of a cheese curd.     

Effect of starter cultures on the physico‐chemical and biochemical features in ewe cheese made with extracts from flowers of Cynara cardunculus L

The effect of starter cultures on the physico‐chemical and biochemical features of Los Pedroches cheese made by using Cynara Cardunculus L extracts as vegetable coagulant was assessed. Specimens of cheese containing no starter (vats A and C) and others to which a lactic acid starter was added (vats B and D) were stored either under typical farmhouse ripening conditions (vats A and B) or in a controlled chamber at 14°C and 80% relative humidity (vats C and D) for 90 days. The addition of starter cultures and the ripening conditions where found to have no appreciable effect on the moisture, fat, protein, ash, NaCl, Ca and P contents of the cheese, nor on its water activity; however, it resulted in increased lactic acid contents and lower pH values relative to control specimens throughout ripening. The protein breakdown of the cheeses was assessed in terms of soluble nitrogen, non‐protein nitrogen, aminoacid nitrogen, ammonia nitrogen, soluble tyrosine and tryptophan, and urea‐PAGE. Proteolysis was generally more marked and rapid in cheese containing the lactic starter, the NPN, NH₃ ‐N and soluble Tyr and Trp contents of which were significantly higher than those of the cheese obtained with no starter. The PAGE technique revealed no appreciable differences between the four cheese batches studied; however, it showed the cheese to be more resistant to β‐CN hydrolysis than to α_s‐CN hydrolysis. In general, the flavour and aroma scores were higher in batches obtained with starter cultures and ripened under controlled conditions. © 1999 Society of Chemical Industry     

Proteolysis in reduced sodium Kefalograviera cheese made by partial replacement of NaCl with KCl

Kefalograviera cheeses (five trials) of different sodium content were made from split lots of curd by varying the salting processes, i.e. brine — and dry — salting, with NaCl (control) or a mixture of NaCl/KCl (3:1 or 1:1, w/w basis). The extent and characteristics of proteolysis in the cheeses were monitored during aging by Kjeldahl determination of soluble nitrogen fractions (water-soluble nitrogen [WSN], trichloroacetic acid [TCA]-SN, phosphotungstic acid [PTA]-SN), the cadmium-ninhydrin method for the determination of total free amino acids (FAA), urea-polyacrylamide gel electrophoresis of cheese proteins, followed by densitometric analysis of the α_s1- and β-casein fractions, reverse-phase high-performance liquid chromatography (HPLC) analysis of the water-soluble extracts of cheeses, and ion-exchange HPLC analysis of FAA. The results showed that proteolysis was similar in control and experimental cheeses at all sampling ages, indicating that the partial substitution of NaCl by KCl in the manufacture of Kefalograviera cheese did not significantly influence the extent and characteristics of proteolysis during cheese aging.