不同方法促熟干酪成熟期间微生物及氨基酸研究

以0、15、30、45、60、75、90 d促熟干酪中主要微生物（乳杆菌、乳球菌、肠球菌、微球菌）和游离氨
基酸含量为指标,研究添加复合处理发酵剂和中性蛋白酶促熟干酪成熟过程对游离氨基酸和微生物菌落数的影响。
结果表明,添加处理发酵剂和中性蛋白酶可增加干酪中游离氨基酸含量,添加处理发酵剂可不同程度降低干酪中微
生物菌落数,添加中性蛋白酶对干酪成熟过程中微生物影响不明显（P＞0.05）。     

发酵剂对牦牛乳硬质干酪成熟过程中生物胺的影响

乳酸菌产生物胺的能力具有菌株特异性,因此,为了探究不同种类发酵剂对牦牛乳硬质干酪中生物胺形成的影响,该试验利用高效液相色谱对3种不同发酵剂制作的硬质干酪成熟过程中生物胺进行了测定和分析。结果表明,嗜热和嗜温发酵剂牦牛乳硬质干酪中检测出2-苯乙胺、腐胺、尸胺、组胺和酪胺,混合发酵剂干酪中检测出腐胺、2-苯乙胺、尸胺和酪胺。各生物胺之间呈现正相关性。3种不同发酵剂干酪在1～6个月成熟过程中,其各生物胺整体呈现增加趋势,嗜热、嗜温和混合发酵剂干酪中总生物胺最高含量分别为(448.3±9.6)、(456.8±58.4)、(293±24.5)mg/kg。组胺和酪胺是2种毒性相对高的生物胺,嗜热发酵剂干酪中组胺和嗜温发酵剂干酪中酪胺最高,其最高含量分别为(20.8±7.9)、(92.9±6.7)mg/kg,混合发酵干酪中未检测出组胺,酪胺含量次之,3种不同发酵剂干酪中组胺、酪胺含量均低于推荐安全剂量50 mg/kg和100 mg/kg。这为合理选择发酵剂和控制干酪中生物胺形成提供了依据。     

发酵剂的添加量对半硬质干酪品质影响研究

研究发酵剂的添加量对半硬质干酪品质的影响,以确定其最佳添加量。采用单因素试验设计,发酵剂的添加量(体积比)分别为1%、2%、3%、4%、5%。在其他工艺条件相同的情况下,分别加工一批干酪,然后测定成熟干酪的性能指标。结果表明,随着发酵剂添加量的增加,干酪的感官评定值先升高后降低;干酪的硬度和pH 值逐渐降低;总酸度、pH4.6 可溶性氮含量、游离氨基酸总量和水分含量逐渐增加。以感官评定值为主要指标,结合其他性能指标,确定发酵剂的最佳添加量为2%～3%。     

不同NaCl添加量对牦牛乳硬质干酪成熟过程中生物胺产生的影响

以不同盐添加量（1.0%、1.3%、1.8%和2.3%）牦牛乳硬质干酪为研究对象,分析牦牛乳硬质干酪0～6 个月成熟过程中生物胺的动态变化,并对干酪中产胺微生物进行筛选。结果显示：不同盐添加量牦牛乳硬质干酪中生物胺主要为色胺、β-苯乙胺、尸胺、酪胺和腐胺,未检测到组胺,生物胺积累阶段主要集中在成熟后期。不同盐添加量干酪中色胺含量最低,且在成熟后期含量差异较小。当盐添加量从2.3%减少到1.0%时,干酪中β-苯乙胺含量减少。盐添加量分别为1.0%和1.3%时,干酪中尸胺含量较低,且未检测出腐胺。当盐添加量在1.8%～2.3%时,随着盐添加量的增加,干酪中尸胺和腐胺含量整体呈现增加趋势。不同盐添加量干酪成熟过程中,其酪胺含量范围为3.13～49.81 mg/kg,且盐添加量为1.3%干酪中酪胺含量较高。不同盐添加量干酪中生物胺总量最高为304.18 mg/kg。采用显色培养基筛选出一株产胺微生物,经分子生物学鉴定为Enterococcus durans。     

微生物发酵剂对兔肉脯游离氨基酸含量的影响

将植物乳杆菌（Lactobacillus plantarum）、木糖葡萄球菌（Staphylococcus xylosus）、汉逊德巴利酵母菌（Dabaryomyces hansenii）接种于兔肉糜中,发酵及烘烤制成兔肉脯,研究不同菌种对产品中游离氨基酸含量的影响。结果表明：三菌种单独发酵处理组中,游离氨基酸总量由高到低分别为：葡萄球菌处理组＞酵母菌处理组＞乳酸菌处理组,3 个处理组游离氨基酸总量均高于空白对照组,说明添加发酵剂促进了兔肉脯中蛋白质的分解,且其在兔肉脯中分解蛋白质产生游离氨基酸的能力依次递减;三菌种单独发酵处理组中,各种游离氨基酸的含量有增有减,说明不同发酵剂对蛋白质分解游离氨基酸的影响不同;三菌种混合发酵处理组总游离氨基酸含量及各氨基酸含量均高于空白对照组和单菌处理组,说明三菌种复合发酵分解蛋白质产生游离氨基酸的效果相对最好。     

发酵剂用量对硬质蒙古干酪蛋白质、脂类水解和微观结构的影响

该文通过添加不同发酵剂用量制作了3种硬质蒙古干酪,并对硬质蒙古干酪的理化性质、蛋白质和脂肪水解程度、微观结构等进行了研究。结果表明,随着发酵剂用量增多,除D-乳酸含量外,其他理化指标均随发酵剂用量的增大而减小;粗蛋白质和粗脂肪的含量减少;pH 4.6-可溶性氮（SN）含量和12%三氯乙酸（TCA）-SN含量均增加,表明干酪中蛋白质和脂肪的水解程度增大;游离氨基酸和游离脂肪酸的含量增加,表明干酪的风味物质增多。随着发酵剂用量的增多,干酪酸化速度加快,质地相对松散。最终确定发酵剂使用量为1.0 g/L,在此条件下,脂肪和蛋白质的水解程度适中,游离氨基酸种类丰富,能够满足干酪风味化合物生成的需求。     

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。     

传统中式香肠成熟过程中生物胺的生物控制

以传统中式香肠成熟过程中生物胺含量的变化为研究对象,重点介绍了色胺、腐胺、尸胺、组胺、酪胺的变化。在香肠加工过程中添加发酵剂和复合植物提取物来控制香肠中生物胺的积累,利用高效液相色谱仪检测成熟过程中生物胺的变化。结果表明,添加发酵剂可有效地抑制色胺、腐胺、尸胺、酪胺的生成;复合植物提取物只是在抑制酪胺含量的增加上有显著效果;同时加入发酵剂和复合植物提取物对色胺、尸胺、组胺和酪胺的抑制效果更显著,特别是组胺,在生产结束时含量为0。发酵剂和复合植物提取物对精胺和亚精胺基本没有影响。     

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。     

牦牛乳硬质干酪成熟过程中生物胺与细菌群落结构分析

成熟干酪易产生生物胺,生物胺对人体健康具有影响,因此,为了探究牦牛乳硬质干酪的质量安全性,本实验利用高效液相色谱和高通量测序技术对干酪成熟过程中生物胺和细菌群落结构进行测定和分析。结果表明：牦牛乳硬质干酪中主要生物胺为腐胺、2-苯乙胺、酪胺、组胺和尸胺,在1～6 个月成熟过程中,干酪中各生物胺含量呈现增加趋势。干酪中生物胺含量最高阶段均出现在成熟期5～6 个月。干酪中组胺、酪胺和总生物胺含量分别低于推荐安全剂量50、100 mg/kg和1 000 mg/kg。干酪中游离氨基酸含量与除组胺之外的各生物胺含量、总生物胺含量、成熟时间之间具有显著正相关性（P＜0.01,P＜0.05）,各生物胺含量之间也存在正相关性。不同成熟期干酪包含相同属的微生物,其中链球菌属（Streptococcus）为优势菌属,平均相对丰度为84.63%,明串珠菌属（Leuconostoc）次之,平均相对丰度为6.91%,其他分别为乳杆菌属（Lactobacillus）、拉乌尔菌属（Raoultella）、不动杆菌属（Acinetobacter）、假单胞菌属（Pseudomonas）、肠杆菌属（Enterobacter）和无分类的肠杆菌科（unclassified_Eunnterobacteriaceae）。不同成熟期干酪中各菌属的相对丰度具有差异。本研究可为评估牦牛乳硬质干酪的质量安全性和生物胺形成机理提供理论依据。     

Effects of heating milk and accelerating ripening of low fat Ras cheese on biogenic amines and free amino acids development

Two experiments were conducted to study the effects of heating milk and, consequently, incorporation of whey proteins into cheese curd, fat content, accelerating cheese ripening by attenuated lactobacilli, species of bacteria and method of attenuation on formation of biogenic amines and liberation of free amino acids. Total biogenic amines and total free amino acids increased as ripening period progressed in all cheese treatments. Total biogenic amines and total free amino acids decreased as the fat content was decreased. Heating of 3% fat milk up to 70°C caused a significant (p<0.05) increase in total biogenic amines and total free amino acid concentrations, while raising the temperature of heat treatment up to 75 and 80°C decreased them. However, heating of 2% fat milk up to 75°C caused a definite (p<0.05) increase in total biogenic arnines and total free amino acids; conversely raising heat treatment temperature to 80°C decreased them. These results indicate that there is a positive correlation between total biogenic amines and total free amino acids; moisture and salt contents affected the formation of biogenic amines, while incorporation of whey proteins had no significant effect on biogenic amines development. In a second experiment, addition of attenuated lactobacilli as adjunct bacteria caused a pronounced (p<0.05) increase in free amino acids and biogenic amines. Addition of either freeze- or heat-shocked Lactobacillus helviticus was more effective in promoting the build-up of biogenic amines and free amino acids in the resultant cheeses than in cheeses made with addition of freeze- or heat-shocked Lactobacillus casei, respectively. There was a positive correlation between free amino acids and biogenic amine contents. The type of bacteria and prolongation of ripening period significantly affected the development of biogenic amines. Tyramine was the highest biogenic amine followed by histamine, while spermidine was the lowest.     

Changes in the free amino acid content during ripening of Idiazabal cheese: influence of starter and rennet type

The effect of starter and rennet type on free amino acid release during ripening of Idiazabal cheese was studied. Four batches of cheeses were manufactured depending on the rennet used, commercial calf rennet or artisanal lamb rennet, and the addition or not of starter culture. Cheese samples contained 24 individual free amino acids Leu, Glu, Val and Phe showing the highest contents during the ripening. The results indicated that the release of the free amino acids during ripening was strongly affected by starter added to the cheeses, and that this effect varied markedly with the rennet used for cheesemaking. Total amounts of free amino acids were higher for the cheeses made with commercial calf rennet than for those made with artisanal lamb rennet, regardless of starter addition. Likewise, the highest total free amino acid levels were found in the cheeses made with starter, regardless of the type of rennet used.     

The effect of elevated temperature on ripening of Dutch type cheese

The aim of this study was to explore the effect of elevated temperature (16 °C) on ripening of Dutch-type cheese. Three slices of each cheese block were further divided into three layers. The processes in the control samples of cheese ripening at 10 °C were also monitored. The contents of free amino acids in accelerated cheese were two times higher than were those in control samples. The highest contents of free amino acids were observed in the cores of all slices of cheeses ripening at both temperatures. The contents of tyramine, in layers of the studied slices, reached almost 500 mg kg⁻¹ during 56 days of the experiment. The contents of biogenic amines in the edges grew even higher. Accelerated cheese showed faster equalisation of hardness than did control samples. The increase of temperature by 6 °C can reduce the ripening time in cellars by approximately one half.     

Interrelationships among microbiological, physicochemical, and biochemical properties of Terrincho cheese, with emphasis on biogenic amines

Changes in the microbiological, physicochemical, and biochemical characteristics of Terrincho cheese as represented by native microflora, pH, water activity, soluble nitrogen fractions, free amino acids, and biogenic amines (e.g., ethylamine, dimethylamine, tryptamine, phenylethylamine, putrescine, cadaverine, histamine, tyramine, cystamine, and spermine) during ripening were monitored. Terrincho is a traditional Portuguese cheese manufactured from raw ewe's milk. The main groups of microorganisms (lactococci, lactobacilli, enterococci, pseudomonads, staphylococci, coliforms, yeasts, and molds) were determined following conventional microbiological procedures. Free amino acids and biogenic amines were determined by reverse-phase high-performance liquid chromatography, following extraction from the cheese matrix and derivatization with dabsyl chloride. The total content of free amino acids ranged from 1,730 mg/kg of dry matter at the beginning of the ripening stage to 5,180 mg/kg of dry matter by day 60 of ripening; such an increase was highly correlated with the increase of water-soluble nitrogen in total nitrogen, 12% trichloroacetic acid-soluble nitrogen in total nitrogen, and 5% phosphotungstic acid-soluble nitrogen in total nitrogen throughout ripening. Histamine was consistently present at very low levels, whereas putrescine, cadaverine, and tryptamine were the dominant biogenic amines and increased in concentration during ripening. Ethylamine, tryptamine, phenylethylamine, and cystamine reached maxima by 30 days of ripening and decreased thereafter. Significant correlations between amino acid precursors and corresponding biogenic amines, as well as between biogenic amines and microbial viable numbers, were observed.     

发酵剂对牦牛乳硬质干酪蛋白降解和苦味关系的影响

用甘肃天祝新鲜牦牛乳为原料分别添加嗜温、嗜热和混合发酵剂制作硬质干酪,以pH 4.6-可溶性氮（soluble nitrogen,SN）、12%三氯乙酸氮（trichloroacrtic acid-N,TCA-N）、游离氨基酸（free amino acid,FAA）含量和疏水性肽/亲水性肽（S/Q）为蛋白水解度指标,研究3 种牦牛乳硬质干酪在6 个月内成熟过程中苦味和蛋白质降解之间的关系。结果表明：3 种干酪在成熟过程中pH 4.6-SN、12% TCA-N和FAA含量均呈上升趋势,苦味值与pH 4.6-SN、12% TCA-N和FAA含量成正相关,相关系数分别为0.400、0.412和0.458。3 种干酪成熟过程中S/Q的变化趋势和程度不同,嗜温发酵剂干酪中S/Q呈现降低趋势;嗜热和混合发酵剂干酪中S/Q均呈现先降低后增大的趋势,但在这两种干酪中S/Q的变化程度不同,嗜热发酵剂干酪在1～3 个月S/Q略有降低,在3～6 个月S/Q快速增大,而混合发酵剂干酪正好相反。S/Q与苦味值成极显著正相关（r=0.895）,S/Q可很好地反映干酪中苦味的强弱。而干酪中苦味强弱与蛋白质降解强弱密切相关,对蛋白降解程度越大的发酵剂制作的干酪越容易产生苦味,其中,嗜热发酵剂对干酪蛋白降解程度最大,混合发酵剂次之,嗜温发酵剂最小。     

Effect of temperature on evolution of free amino acid and biogenic amine contents during storage of Azeitão cheese

A study on the evolution of free amino acids and biogenic amines in Azeitão cheese during 4 weeks at different temperatures of storage (4 and 25°C) was performed. Free amino acids and biogenic amines were determined by RP-HPLC with visible detection, following extraction from the cheese and derivatization with dabsyl chloride. The method presented a linear relation between peak area and concentration from 2–200 mg/l. The detection limit value was less than 1.5 mg/l. The average repeatability was less than 4%. The major free amino acids were proline, valine, isoleucine and leucine and the major amines were tyramine, cadaverine and histamine. Room temperature (25°C) promoted a significant increase of the contents of valine, leucine, tyramine and putrescine, expressed as g/kg of dry matter. These two free amino acids and two biogenic amines may serve as indicators of temperatures changes in ripened cheese.     

Influence of starter and nonstarter on the formation of biogenic amine in goat cheese during ripening

Two commercial starters were investigated for their potential ability to decarboxylate amino acids during goat cheese ripening. Two batches of goat cheese were produced with identical pasteurized milk but different starter cultures. One of them contained Lactococcus lactis subsp. lactis and Lactococcus lactis subsp. cremoris and the other Lactococcus lactis subsp. lactis. The amine contents, microbial counts, proteolysis-related parameters, pH, total solids and salt content were studied in raw materials and cheeses. In raw materials, polyamines were the prevailing amines, whereas the main amines in cheeses were putrescine, tryptamine and, in particular, tyramine (94.59 mg/kg). Aerobic mesophilic microorganisms and Lactococcus counts increased throughout ripening, while Enterobacteriaceae were no longer detectable in cheese after 30 days of ripening. Amine concentration rose during cheese ripening in both batches. Moreover, the decarboxylase activity of microorganisms isolated from samples during cheese ripening was assayed and discussed.