乳酸菌的筛选及其对羊肉干发酵香肠品质特性的影响
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摘要
从内蒙古不同地区传统肉肠中分离筛选乳酸菌，并对其加工及功能特性进行研究，从而优选出适合用做发酵香肠发酵剂的菌株。将筛选得到的乳酸菌单独或与木瓜蛋白酶共同用于羊肉干发酵香肠的加工，测定发酵香肠加工及贮存过程中微生物、理化及风味、感官等特性的变化，探讨乳酸菌及木瓜蛋白酶对羊肉干发酵香肠品质特性的影响及其机理。结果表明：
从内蒙肉肠中分离、筛选得到8株符合肉制品发酵剂特性的菌株,经形态、生理生化特性鉴定及16SrDNA序列分析，证明8株菌为：5株植物乳杆菌、3株弯曲乳杆菌。筛选菌株在不同的温度、pH条件下均具有较强的生长能力，其中X3-2B、X3-3B、X3-4B及X3-5B在25、30、37℃条件下生长能力显著强于其它温度（P<0.05），在pH4.5、5.5及6.5条件下X3-2B、X3-3B、X3-4B及X3-5B生长能力极显著强于其它菌株（P<0.01）。在不同的培养基中均能够在24h内使pH值降低至4.2以下。X3-2B及X3-3B在人工胃液中的存活率显著高于其它菌株（P<0.05）；X3-2B及X3-5B的耐胆盐能力最强（P<0.05）；X3-2B的胆固醇降解率为68.92%，降解能力显著高于其它菌株（P<0.05）；X3-2B及X3-5B对大肠杆菌及金黄色葡萄球菌表现出较强的抑制作用。综合以上特性，X3-2B适合作为发酵剂用于发酵香肠的加工。
将植物乳杆菌X3-2B单独或与木瓜蛋白酶共同用于羊肉干发酵香肠的加工，未加乳酸菌组做对照。在羊肉干发酵香肠的加工及贮存过程中，各组乳酸菌数在发酵结束时均达到了108cfu/g，试验组显著高于对照组（P<0.05）；试验组细菌总数显著低于对照组（P<0.05），乳酸菌的添加明显抑制了细菌总数的增加。乳酸菌明显加快了试验组pH值、Aw值及亚硝酸盐的下降速率（P<0.05），减缓了挥发性盐基氮的增长速率（P<0.05）；乳酸菌的添加增加了香肠的硬度，降低了其弹性；试验组发酵香肠的色差影响参数显著高于对照组（P<0.05），乳酸菌的添加显著改善了产品的色泽。试验组间上述指标均存在一定的差异，但差异不显著。说明乳酸菌对羊肉干发酵香肠的理化特性产生了显著影响，而木瓜蛋白酶的添加对其影响不显著。
在羊肉干发酵香肠干燥及成熟阶段试验组有新的小分子蛋白条带出现，贮藏过程中则有小分子蛋白条带消失；在加工及贮藏过程中试验组游离氨基酸的释放量明显高于对照组，且木瓜蛋白酶组高于乳酸菌组；各组羊肉干发酵香肠中检出风味物质的种类及含量为木瓜蛋白酶组>乳酸菌组>对照组；试验组感官评定得分明显高于对照组。说明乳酸菌X3-2B的添加对羊肉干发酵香肠风味特性的形成有明显的促进作用，而乳酸菌与木瓜蛋白酶共同作用使促进作用更加明显。
To screen out the potential strains used as starter culltures for fermented sausage, theprocessing and functional characteristics of lactic acid bacteria isolated from traditionalsausages of different areas in Inner Mongolia were studied. After producing mutton dryfermented sausage by single selected strains or the combination of selected strain andpapain, the microbial, physiochemical, flavor and sensory properties of fermented sausageunder processing and storage conditions were tested. Moreover, the influence oflactobacilli and papain on the quality of mutton dry fermetned sausage was evaluated. Theresults showed that:
Eight lactobacilli strains which possess the required fermented properties of startercultures used for meat products were screened out from Inner Mongolia traditionalsausages. Through morphous, physiological, biochemical characteristics and16S rDNAsequence analysis, these eight strains were identified as five strains of Lactobacillusplantarum, three strains of Lactodacillus curvature All the selected strains can expressstrong growth adaptability under different conditions with various temperature and pHvalues. The growth of X3-2B, X3-3B, X3-4B and X3-5B at25,30,37℃were significantlyhigher than those at other temperatures （P<0.05）. Moreover, the growth of these fourstrains under pH4.5,5.5and6.5were significantly higher than that of other strains（P<0.01）. All the selected strains can reduce the pH value of various culture media to4.2within24h. The survival rate of X3-3B and X3-2B in artifical gastric juice weresignificantly higher than that of other strains （P<0.05）. The bile salt resistance ability ofX3-2B and X3-5B were the highest. The cholestero removal rate of X3-2B wassignificantly higher than that of other strains, with68.92%. Moreover, X3-2B and X3-5Bexpressed strong inhibitory effect on the growth of E. coli and Staphylococcus aureus.Therefore, it is concluded that X3-2B is suitable to act as starter culture for the productionof fermented sausage. Mutton dry fermented sausage was produced by X3-2B or thecombination of X3-2B and papain, with the products without LAB starter culture as controlgroup. The results showed that the viable count of lactobacilli in trial groups weresignificantly higher than that of control group during processing and storage of mutton dryfermented sausage. However, their total viable counts were significantly lower than that of control group （P<0.05）. pH value, Aw, nitrite content and TVBN of trial groups weresignificantly lower than those of control group. But their texture and color weresignificantly higher （P<0.05）. There were no significant difference between the group ofX3-2B and X3-2B with papain. Thus X3-2B had a significant impact on thephysicochemical properties of mutton dry fermented sausage, and adding papain had nosignificant effect on it.
There were small molecule protein bands appeared in trail groups during drying andmaturity stage, and small molecular protein bands disappeared during storage; Releaseamount of free amino acid of trial groups were significantly higher than control groupduring the processing and storage, and X3-2B+papain group was higher than X3-2Bgroup; Species and contents of flavor substances in each group were：X3-2B+papaingroup> X3-2B group> control group; sensory evaluation score of trial groups weresignificantly higher than that of control group.It was concluded that the addition oflactobacilli could obviously promote the formation of mutton flavor characteristics of dryfermented sausage, and papain can enhance this effect.
从内蒙肉肠中分离、筛选得到8株符合肉制品发酵剂特性的菌株,经形态、生理生化特性鉴定及16SrDNA序列分析，证明8株菌为：5株植物乳杆菌、3株弯曲乳杆菌。筛选菌株在不同的温度、pH条件下均具有较强的生长能力，其中X3-2B、X3-3B、X3-4B及X3-5B在25、30、37℃条件下生长能力显著强于其它温度（P<0.05），在pH4.5、5.5及6.5条件下X3-2B、X3-3B、X3-4B及X3-5B生长能力极显著强于其它菌株（P<0.01）。在不同的培养基中均能够在24h内使pH值降低至4.2以下。X3-2B及X3-3B在人工胃液中的存活率显著高于其它菌株（P<0.05）；X3-2B及X3-5B的耐胆盐能力最强（P<0.05）；X3-2B的胆固醇降解率为68.92%，降解能力显著高于其它菌株（P<0.05）；X3-2B及X3-5B对大肠杆菌及金黄色葡萄球菌表现出较强的抑制作用。综合以上特性，X3-2B适合作为发酵剂用于发酵香肠的加工。
将植物乳杆菌X3-2B单独或与木瓜蛋白酶共同用于羊肉干发酵香肠的加工，未加乳酸菌组做对照。在羊肉干发酵香肠的加工及贮存过程中，各组乳酸菌数在发酵结束时均达到了108cfu/g，试验组显著高于对照组（P<0.05）；试验组细菌总数显著低于对照组（P<0.05），乳酸菌的添加明显抑制了细菌总数的增加。乳酸菌明显加快了试验组pH值、Aw值及亚硝酸盐的下降速率（P<0.05），减缓了挥发性盐基氮的增长速率（P<0.05）；乳酸菌的添加增加了香肠的硬度，降低了其弹性；试验组发酵香肠的色差影响参数显著高于对照组（P<0.05），乳酸菌的添加显著改善了产品的色泽。试验组间上述指标均存在一定的差异，但差异不显著。说明乳酸菌对羊肉干发酵香肠的理化特性产生了显著影响，而木瓜蛋白酶的添加对其影响不显著。
在羊肉干发酵香肠干燥及成熟阶段试验组有新的小分子蛋白条带出现，贮藏过程中则有小分子蛋白条带消失；在加工及贮藏过程中试验组游离氨基酸的释放量明显高于对照组，且木瓜蛋白酶组高于乳酸菌组；各组羊肉干发酵香肠中检出风味物质的种类及含量为木瓜蛋白酶组>乳酸菌组>对照组；试验组感官评定得分明显高于对照组。说明乳酸菌X3-2B的添加对羊肉干发酵香肠风味特性的形成有明显的促进作用，而乳酸菌与木瓜蛋白酶共同作用使促进作用更加明显。
To screen out the potential strains used as starter culltures for fermented sausage, theprocessing and functional characteristics of lactic acid bacteria isolated from traditionalsausages of different areas in Inner Mongolia were studied. After producing mutton dryfermented sausage by single selected strains or the combination of selected strain andpapain, the microbial, physiochemical, flavor and sensory properties of fermented sausageunder processing and storage conditions were tested. Moreover, the influence oflactobacilli and papain on the quality of mutton dry fermetned sausage was evaluated. Theresults showed that:
Eight lactobacilli strains which possess the required fermented properties of startercultures used for meat products were screened out from Inner Mongolia traditionalsausages. Through morphous, physiological, biochemical characteristics and16S rDNAsequence analysis, these eight strains were identified as five strains of Lactobacillusplantarum, three strains of Lactodacillus curvature All the selected strains can expressstrong growth adaptability under different conditions with various temperature and pHvalues. The growth of X3-2B, X3-3B, X3-4B and X3-5B at25,30,37℃were significantlyhigher than those at other temperatures （P<0.05）. Moreover, the growth of these fourstrains under pH4.5,5.5and6.5were significantly higher than that of other strains（P<0.01）. All the selected strains can reduce the pH value of various culture media to4.2within24h. The survival rate of X3-3B and X3-2B in artifical gastric juice weresignificantly higher than that of other strains （P<0.05）. The bile salt resistance ability ofX3-2B and X3-5B were the highest. The cholestero removal rate of X3-2B wassignificantly higher than that of other strains, with68.92%. Moreover, X3-2B and X3-5Bexpressed strong inhibitory effect on the growth of E. coli and Staphylococcus aureus.Therefore, it is concluded that X3-2B is suitable to act as starter culture for the productionof fermented sausage. Mutton dry fermented sausage was produced by X3-2B or thecombination of X3-2B and papain, with the products without LAB starter culture as controlgroup. The results showed that the viable count of lactobacilli in trial groups weresignificantly higher than that of control group during processing and storage of mutton dryfermented sausage. However, their total viable counts were significantly lower than that of control group （P<0.05）. pH value, Aw, nitrite content and TVBN of trial groups weresignificantly lower than those of control group. But their texture and color weresignificantly higher （P<0.05）. There were no significant difference between the group ofX3-2B and X3-2B with papain. Thus X3-2B had a significant impact on thephysicochemical properties of mutton dry fermented sausage, and adding papain had nosignificant effect on it.
There were small molecule protein bands appeared in trail groups during drying andmaturity stage, and small molecular protein bands disappeared during storage; Releaseamount of free amino acid of trial groups were significantly higher than control groupduring the processing and storage, and X3-2B+papain group was higher than X3-2Bgroup; Species and contents of flavor substances in each group were：X3-2B+papaingroup> X3-2B group> control group; sensory evaluation score of trial groups weresignificantly higher than that of control group.It was concluded that the addition oflactobacilli could obviously promote the formation of mutton flavor characteristics of dryfermented sausage, and papain can enhance this effect.
引文
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