乳酸菌分离筛选及其对新型半硬质干酪品质特性影响
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摘要
试验从国内外12份传统干酪及奶豆腐制品中分离乳酸菌，并按照新型半硬干酪发酵剂的要求对乳酸菌进行综合筛选。从样品中分离出符合乳酸菌特性的菌株37株，其中4株适合作为干酪发酵剂的乳酸菌。经常规生理生化、API50CH试剂盒和16SrDNA鉴定法相结合将这4株菌进行种属的鉴定，鉴定结果为菌株K2为植物乳杆菌（Lactobacillus. plantarum）、菌株D1为清酒乳杆菌（Lactobacillus. sakei）、菌株G1为屎肠球菌（Enterococcus.faecium）、菌株F1为乳酸链球菌属乳脂亚种（Lactococcus.lactis subsp. cremoris）。
在传统Cheddar干酪加工工艺的基础上，通过单因素对比试验及正交试验对干酪的生产工艺进行改良优化，确定了新型半硬干酪生产的最佳参数。这些参数包括：发酵剂乳酸菌的最佳发酵温度32℃，凝乳酶（活力1070IMCU/g）最佳用量3g/100L牛乳，凝乳温度32℃，堆酿温度42℃，盐水浸泡时盐水的浓度16%、盐水与干酪的比例2：1、浸泡时间2.5h。在此工艺参数下生产的半硬干酪得率在10%以上，水分含量在50%左右，且干酪的咀嚼性及感官指标较优。
将分离筛选出的4株乳酸菌进行研究其单一生长和混合生长的生长特性和发酵牛乳的特性，研究结果为K2菌株与F1菌株、D1菌株与G1菌株混合培养时有相互促进的作用。当K2：F1=5：1用量为5%或D1：G1=1：1用量为2%时生产出的干酪特性较好。
以优选出的2组组合按最适比例及添加量制作的干酪为试验组，以不加发酵剂的干酪为对照组，通过研究3组干酪在成熟过程中的感官指标、质构指标、理化指标及微生物指标以研究这两组发酵剂乳酸菌对干酪成熟特性的影响。结果显示使用发酵剂乳酸菌的干酪对干酪的成熟特性产生显著的影响。随着干酪的成熟发酵剂组干酪中的水分含量、水分活度、酸度、蛋白质含量、可溶性氮含量、游离氨基酸、脂肪含量、游离脂肪酸、质构及色差等指标均呈现显著的影响。并且菌株组合之间差异显著。根据这些特性，最终选择其中特性优良的组合K2F1作为干酪的发酵剂乳酸菌。这一组组合能显著的提高干酪的成熟特性，使得干酪的成熟期缩短，这对于干酪工业化生产有十分重要的意义。
The lactic acid bacteria were isolated and identified from twelve cheesesamples,which were collected from outside country and inner Mongolia. The lactic acidbacteria were screened and used as cheese starter culture of Chinese tastescheese.Through studying the value changes of physico-chemical characterization andmicroorganism.The results were as follow:37strains of lactic acid bacteria were screenedfrom the samples, Among them4strains were better by detecting some fermentedcharacteristics(for example acid production,salt tolerance,sensory evaluation ofcheese,texture of cheese,the moisture of the cheese,whey output, fermentedcharacterization of skim milk during14hours). And then the four strains were identifiedby traditional physio-chemical and API50CH and16SrDNA molecular methods, Theresults indicated that K2strain and D1strain were Lactobacillus.the K2strain isLactobacillus.Plantarum and the D1strain is Lactobacillus. Sakei,and the other twostrains G1and F1were Lactococcus，and the G1strain is Enterococcus.faecium,the F1strain is Lac. lactis subsp. Cremoris.
The influence of4lactic acid bacteria strains were used as starter according todifferent proportion and amount to processed semi-hard cheese, according to assayingviscosity and acid conditions in fermented milk.and then the best proportion and amountwere decided,which were the proportion K2and F1=5:1was much higher than that of K2or F1alone at the time of stable stage, and the amount was5%,and the same time anothergroup was selected,that was D1and G1=1:1was much higher than that of D1or G1alone at the time of stable stage,and the amount was2%,and then the two best groups ofthe four strains on the characeristics of semi-hard cheese were discussed.the two groupstrains had been added in cheese.Testing various physical,chemical and microbialindicators during cheese ripening process,and result showed that the addition of staterstrains had changed the general characteristics of semi-hard cheese,such as the contentof moisture and the value of pH and the texture and pH4.6SN and12%TCA SN and thecontents of total free amino acid and the individual free amino acid in the experimentalcheese were significantly different from the control(P<0.05). the pH4.6SN and12%TCA SN and the contents of total free amino acid and the individual free amino acid increase obviously during the ripening period of the cheese.and the pH value reducedrapidly during2months in the two group experimental cheeses,lately it rised slowly andkept invariable.The number of microorganisms decreased rapidly early and kept stableeventually.In addition,the group of K2F1increased the hardness and reduced thespringing of products significantly during the ripening period of the cheese,and thecolour of cheese was significant effected by the starter culture of K2F1during theripening period.As the result,the strains of K2F1can be used as starter in cheese.
在传统Cheddar干酪加工工艺的基础上，通过单因素对比试验及正交试验对干酪的生产工艺进行改良优化，确定了新型半硬干酪生产的最佳参数。这些参数包括：发酵剂乳酸菌的最佳发酵温度32℃，凝乳酶（活力1070IMCU/g）最佳用量3g/100L牛乳，凝乳温度32℃，堆酿温度42℃，盐水浸泡时盐水的浓度16%、盐水与干酪的比例2：1、浸泡时间2.5h。在此工艺参数下生产的半硬干酪得率在10%以上，水分含量在50%左右，且干酪的咀嚼性及感官指标较优。
将分离筛选出的4株乳酸菌进行研究其单一生长和混合生长的生长特性和发酵牛乳的特性，研究结果为K2菌株与F1菌株、D1菌株与G1菌株混合培养时有相互促进的作用。当K2：F1=5：1用量为5%或D1：G1=1：1用量为2%时生产出的干酪特性较好。
以优选出的2组组合按最适比例及添加量制作的干酪为试验组，以不加发酵剂的干酪为对照组，通过研究3组干酪在成熟过程中的感官指标、质构指标、理化指标及微生物指标以研究这两组发酵剂乳酸菌对干酪成熟特性的影响。结果显示使用发酵剂乳酸菌的干酪对干酪的成熟特性产生显著的影响。随着干酪的成熟发酵剂组干酪中的水分含量、水分活度、酸度、蛋白质含量、可溶性氮含量、游离氨基酸、脂肪含量、游离脂肪酸、质构及色差等指标均呈现显著的影响。并且菌株组合之间差异显著。根据这些特性，最终选择其中特性优良的组合K2F1作为干酪的发酵剂乳酸菌。这一组组合能显著的提高干酪的成熟特性，使得干酪的成熟期缩短，这对于干酪工业化生产有十分重要的意义。
The lactic acid bacteria were isolated and identified from twelve cheesesamples,which were collected from outside country and inner Mongolia. The lactic acidbacteria were screened and used as cheese starter culture of Chinese tastescheese.Through studying the value changes of physico-chemical characterization andmicroorganism.The results were as follow:37strains of lactic acid bacteria were screenedfrom the samples, Among them4strains were better by detecting some fermentedcharacteristics(for example acid production,salt tolerance,sensory evaluation ofcheese,texture of cheese,the moisture of the cheese,whey output, fermentedcharacterization of skim milk during14hours). And then the four strains were identifiedby traditional physio-chemical and API50CH and16SrDNA molecular methods, Theresults indicated that K2strain and D1strain were Lactobacillus.the K2strain isLactobacillus.Plantarum and the D1strain is Lactobacillus. Sakei,and the other twostrains G1and F1were Lactococcus，and the G1strain is Enterococcus.faecium,the F1strain is Lac. lactis subsp. Cremoris.
The influence of4lactic acid bacteria strains were used as starter according todifferent proportion and amount to processed semi-hard cheese, according to assayingviscosity and acid conditions in fermented milk.and then the best proportion and amountwere decided,which were the proportion K2and F1=5:1was much higher than that of K2or F1alone at the time of stable stage, and the amount was5%,and the same time anothergroup was selected,that was D1and G1=1:1was much higher than that of D1or G1alone at the time of stable stage,and the amount was2%,and then the two best groups ofthe four strains on the characeristics of semi-hard cheese were discussed.the two groupstrains had been added in cheese.Testing various physical,chemical and microbialindicators during cheese ripening process,and result showed that the addition of staterstrains had changed the general characteristics of semi-hard cheese,such as the contentof moisture and the value of pH and the texture and pH4.6SN and12%TCA SN and thecontents of total free amino acid and the individual free amino acid in the experimentalcheese were significantly different from the control(P<0.05). the pH4.6SN and12%TCA SN and the contents of total free amino acid and the individual free amino acid increase obviously during the ripening period of the cheese.and the pH value reducedrapidly during2months in the two group experimental cheeses,lately it rised slowly andkept invariable.The number of microorganisms decreased rapidly early and kept stableeventually.In addition,the group of K2F1increased the hardness and reduced thespringing of products significantly during the ripening period of the cheese,and thecolour of cheese was significant effected by the starter culture of K2F1during theripening period.As the result,the strains of K2F1can be used as starter in cheese.
引文
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