乳酸乳球菌乳脂亚种高产丁二酮的条件优化

分别研究了10种不同因素对乳酸乳球菌乳脂亚种产生丁二酮质量分数的影响.结果表明,各因素产生丁二酮最高量的条件:培养温度为37℃;凝乳后3 h;后熟12 h;培养基pH值调至为5.6;培养基中添加柠檬酸氢二铵的质量浓度为0.15%:培养基中添加Vc的质量浓度为0.3%.培养基中添加金属离子的质量浓度分别为:Mg2+为O.05%,Cu2+为O.04%,Mn2+为O.001%;培养基中添加甘氨酸的质量浓度为1.7%(均为质量分数);培养基中添加碳水化舍物的质量浓度分别为:葡萄糖为1%,蔗糖为1.5%(均为质量分数);随着基质浓度增加丁二酮产量也随之增加.     

乳酸乳球菌乳脂亚种冷冻干燥保护剂优化及其贮藏稳定性

采用单因素试验、Plackett-Burman试验设计和Box-Behnken响应面法,筛选和优化乳酸乳球菌乳脂亚种KLDS4.0326真空冷冻干燥保护剂工艺,并分析冻干菌粉在不同温度下的贮藏稳定性。结果表明：脱脂乳粉、海藻糖和甘油含量是影响菌体细胞存活率的关键因素;以菌体细胞存活率为响应值,对3 个因素进行优化分析,保护剂最佳配比为：脱脂乳粉10.77 g/100 mL、海藻糖7.81 g/100 mL、甘油3.31 g/100 mL,此条件下菌体细胞存活率为（87.43±1.62）%,与理论预测值接近。在4 ℃和25 ℃条件下保藏12 个月后,菌体细胞存活率分别为40.63%和8.67%,冻干菌粉在4 ℃条件下具有更高贮藏稳定性。     

Subspecies-Specific Nested PCR Assay for Detection of Lactococcus lactis spp. lactis and spp. cremoris

A nested PCR-based assay composed of Lactococcus lactis species-specific primers for the nest 1 amplification and subspecies-specific primers for the nest 2 amplification was validated with the identified strains of L. lactis isolated from dairy and nondairy sources and positive and negative control strains. Forward and reverse primer set was designed for nest 1 amplification targeting the conserved housekeeping gene yueF encoding nonproteolytic protein from peptidase family M16 of L. lactis. Amplicons of 447 bp of yueF were subjected for nest 2 amplification producing amplicons of 372 bp. The designed outer primer set for nest 1 amplification was observed to be specific to L. lactis because the DNA from other bacteria could not be amplified and the inner primer set for nest 2 amplification was found to be specific for the detection of ssp. lactis and cremoris of L. lactis.     

乳酸乳球菌胞外多糖磷酸化的工艺优化

采用单因素和正交试验,对乳酸乳球菌胞外多糖的磷酸化工艺进行研究,探讨磷酸盐用量、反应温度、反应时间、反应pH值对乳酸乳球菌胞外多糖最终PO43-接枝量的影响。所得的乳酸乳菌球菌胞外多糖磷酸化的工艺优化条件为：胞外多糖与磷酸盐质量比为6:1、温度90℃、时间4h、pH6.0,此条件下所得PO43-的接枝量为1.639mg/g。     

乳酸乳球菌乳酸亚种高产胆盐水解酶发酵条件的优化研究

本文利用从藏灵菇中筛选的产胆盐水解酶的乳酸乳球菌乳酸亚种(Lactococcuslactissubsp.lactis)研究提高酶活力的环境因素。针对主要影响胆盐水解酶合成的四个因素,采用四因素三水平L9(34)正交试验确定了高产胆盐水解酶优化发酵条件:葡萄糖添加量为3%、大豆蛋白胨添加量为2%、发酵温度为37℃、接种量为2%。在优化条件下,胆盐水解酶活力是优化前的11.84倍。     

镉胁迫下泡菜乳酸乳球菌的形态变化

通过扫描电镜和透射电镜分别观察不同质量浓度水平的Cd2+对泡菜乳酸乳球菌（Lactococcus lactis subsp.lactis）细胞的影响,扫描电镜结果显示：Cd2+质量浓度在0、10 mg/L时,泡菜乳酸乳球菌呈椭圆形、表面光滑、菌体生长繁殖旺盛,随着Cd2+质量浓度的增加菌体细胞表面产生白色颗粒状物质、菌体细胞存活数量大幅下降（OD600 nm值由1.336下降到0.515）。当添加200 mg/L Cd2+时,几乎没有见到明显的菌体、显示有少量棱形晶状物。透射电镜结果显示：当Cd2+质量浓度为0～50 mg/L时泡菜乳酸乳球菌结构完整、细胞内容物分布均、菌体生长较为正常,当菌体暴露于100、200 mg/L Cd2+时菌体细胞出现异常现象,如细胞破裂、内容物从薄膜穿孔中释放、质壁分离等。两类电镜结果均表明：在低质量浓度Cd2+（≤50 mg/L）胁迫下,对泡菜乳酸乳球菌的生长几乎不产生影响,添加Cd2+质量浓度上升到100、200 mg/L时泡菜乳酸乳球菌正常生长受到抑制。     

一株乳酸乳球菌所产细菌素的生物学特性

对乳酸乳球菌KLDS4.0326所产类细菌素进行分离纯化,并对细菌素的部分生物学特性进行研究。在排除酸性产物和过氧化氢的干扰后,菌株的无细胞发酵上清液对金黄色葡萄球菌和枯草芽孢杆菌等革兰氏阳性细菌以及大肠杆菌、沙门氏菌等革兰氏阴性细菌具有显著的抑制作用;经胰蛋白酶、胃蛋白酶、木瓜蛋白酶和蛋白酶K处理后,抑菌活性降低,表明抑菌成分为蛋白类物质;该细菌素具有良好的热稳定性,在酸性条件下抑菌活性稳定,并且具有较广的抑菌谱,能够抑制多种革兰氏阳性细菌和革兰氏阴性细菌。     

乳酸乳球菌胞外多糖的分离及其体外抗氧化活性研究

本实验对乳酸乳球菌胞外多糖(EPS)的离心除菌、提取、粗胞外多糖中蛋白质的去除进行研究,同时探究胞外多糖的体外抗氧化活性。结果表明：6000r/min 离心15min;温度15℃;旋转蒸发浓缩到原体积的1/4;醇沉最佳条件为３倍糖溶液体积的90% 乙醇沉淀12h;选择TCA 沉淀蛋白,TCA 的最佳浓度为10%;胞外多糖有显著的体外抗氧化性。     

Effects of yeast fermentation on transforming the volatile compounds of unsalted pork hydrolysate

This study investigated the possibility of transforming unsalted pork hydrolysate into a liquid seasoning by studying volatile compound production through yeast fermentation. The yeasts used included a typical soya sauce yeast and three wine yeasts. The yeasts inoculated in the glucose supplementary pork hydrolysate increased by approximately 2.0 log of CFU mL⁻¹ within 24 hours at 30°C without adding salt. Yeast fermentation largely depleted the abundant aldehydes (hexenal) present in unfermented pork hydrolysate to trace levels with the formation of ethanol, corresponding alcohols and carboxylic acids. The wine yeasts showed higher production of fruity esters such as ethyl acetate and isoamyl acetate, whereas the soya sauce yeast produced more ketones such as acetone and 2-heptanone. This study revealed a potential method of producing a value-added meat hydrolysate from pork by-product.     

Use of Taiwanese ropy fermented milk (TRFM) and Lactococcus lactis subsp. cremoris isolated from TRFM in manufacturing of functional low-fat cheeses

Abstract: The purpose of this study was to manufacture new functional low‐fat cheeses using Taiwanese ropy fermented milk (TRFM) and Lactococcus lactis subsp. cremoris strains isolated from TRFM. After 28 d of ripening and storage, the viable populations of lactic acid bacteria (LAB) in the low‐fat cheeses made with L. lactis subsp. cremoris TL1 (TL1), L. lactis subsp. cremoris TL4 (TL4), and TRFM still maintained above 10⁸ CFU/g. The low‐fat cheeses made with TL1 and TRFM showed higher moisture contents than the cheeses made with TL4, full‐fat, and low‐fat cheese controls. The low‐fat cheeses made with TL1 and TL4 had higher customer preferential scores similar to full‐fat cheese control in the sensory evaluation. Additionally, the low‐fat cheeses fermented with TL1, TL4, and TRFM for 4 h had higher 1,1‐diphenyl‐2‐picrylhydrazyl (DPPH) free radical‐scavenging and ferrous ion‐chelating abilities than the cheeses fermented with the starters for 8 h, full‐fat, and low‐fat cheese controls. A better angiotensin‐converting enzyme (ACE) inhibition activity was also observed in the low‐fat cheeses made with TL1, TL4, and TRFM than that in the full‐fat and low‐fat cheese controls during ripening and storage period. Practical Application: As health‐conscious consumers continue to seek low‐fat alternatives in their diets, there remain strong interests for the dairy industry to develop low‐fat cheeses to meet the demands. This study clearly demonstrated that the low‐fat cheeses fermented with TL1 for 4 h showed a better overall acceptability and possessed antioxidative abilities and ACE inhibitory activities than other cheeses tested in this study. By improving its flavor and investigating the possible mechanisms of its functionalities in the future, this low‐fat cheese might possibly be commercialized and give a positive impact on cheese consumption in the future.     

乳脂乳球菌复配发酵乳贮藏期品质变化研究

探讨了分离自新疆酸奶疙瘩的一株乳脂乳球菌,在乳制品应用中具有的潜在益生价值。通过测定多肽、滴定酸度、丁二酮、乙醛、黏度、活菌数等指标,研究了乳脂乳球菌复配菌株发酵乳在贮藏期间的发酵特性。结果表明,在贮藏期发酵乳各指标均显著高于对照组,酸度升高了29.59 °T,后酸化能力较强。丁二酮含量在整个贮藏过程中较稳定,乙醛、多肽、黏度、胞外多糖的含量在贮藏7 d后达到最大值。多肽含量最高达3.34 mg/mL,胞外多糖含量与测定的黏度值呈现正相关。活菌数呈下降趋势,贮藏21 d后,为7.8×107 CFU/mL,仍高于发酵乳标准中活菌数1×106 CFU/mL的要求。     

乳酸乳球菌乳酸亚种产丁二酮的优化研究

分别研究了10种不同因素对乳酸乳球菌乳酸亚种产生丁二酮含量的影响。实验结果表明,各因素产生丁二酮最高量的条件分别为:培养温度为37℃,凝乳后0h,后熟12h,培养基pH调节为5.6,培养基中添加柠檬酸的量为0.15%;培养基中添加Vc的量为0.01%,培养基中添加金属离子的量分别为Mg2+0.03%、Cu2+0.02%、Mn2+0%,培养基中添加甘氨酸的量为0.2%,培养基中添加碳水化合物的量分别为葡萄糖为0%、蔗糖为0%;随着基质浓度增加丁二酮产量也随之增加。     

乳酸菌胞外多糖的纯化及对小鼠血清和肝组织抗氧化性的影响

本实验对乳酸乳球菌乳亚种胞外多糖的纯化工艺及其抗氧化活性进行研究。纯化结果表明,选择DA201-C树脂,调节胞外多糖的pH值为3.5,在45℃条件下,树脂用量为2%(V/V),静态吸附20h后糖保留率为81.56%,蛋白脱除率为72.09%,脱色率为81.69%。通过动物实验,检测小鼠血清和肝组织中的过氧化氢酶(CAT)活力、丙二醛(MDA)含量、超氧化物歧化酶(SOD)活力可知,灌胃胞外多糖组的小鼠与对照组小鼠相比,抗氧化能力显著性提高。     

乳脂乳球菌CH5生长特性的研究

本文以农家干酪生产使用的乳脂乳球菌(Lactococcus lactis ssp．cremoris)为研究对象，从细菌总数(CFU)，pH值和滴定酸度三个指标对本实验室分离得到的乳脂乳球菌CH5在脱脂乳中的生长特性和4℃下保存过程中的活力变化进行了研究。结果表明乳脂乳球菌CH5在脱脂乳中的延滞期短，能较快进入对数生长期，6h时进入稳定期，活菌数达109CFU／ml，Tdg约为1．03h，Tda约为2．87h，这表明乳脂乳球菌CH5细胞增长速度快于产酸速率。乳脂乳球菌CH5在4℃下保存时，稳定性较好，前11d活菌数和最大产酸速率(μacid)基本保持不变，此后逐渐下降，表明CH5发酵剂在前11d的活力比较稳定，可以确定生产农家干酪的CH5母发酵剂的最佳传代时间为11d，冷却保藏第3d为最佳生产使用时间。     

乳酸乳球菌发酵液中乳链菌肽的分离纯化

对乳酸乳球菌乳酸亚种SQ80随pH的改变吸附乳链菌肽的规律进行了研究，pH6．5时吸附率达91．5％，pH3以下吸附率为0。通过调整pH，使乳酸乳球菌选择性地吸附、释放乳链菌肽，达到了较好的分离纯化效果，HPLC检测显示单一尖峰，乳链菌肽回收率58．2％，纯化倍数75。     

The effect of lactose, NaCl and an aero/anaerobic environment on the tyrosine decarboxylase activity of Lactococcus lactis subsp. cremoris and Lactococcus lactis subsp. lactis

The aim of this work was to study, under model conditions, combined effects of the concentration of lactose (0-1% w/v), NaCl (0-2% w/v) and aero/anaerobiosis on the growth and tyramine production in 3 strains of Lactococcus lactis subsp. lactis and 2 strains of L. lactis subsp. cremoris. The levels of the factors tested were chosen with respect to the conditions which can occur during the real process of natural cheese production, including the culture temperature (10 ± 1 °C). In all strains tested, tyrosine decarboxylation was most influenced by NaCl concentration; the highest production of tyramine was obtained within the culture with the highest (2% w/v) salt concentration applied. Two of the strains L. lactis subsp. lactis produced tyramine only in broth with the highest NaCl concentration tested. In the remaining 3 strains of L. lactis, tyramine was detected under all conditions applied. The tested concentration of lactose and aero/anaerobiosis had a less significant effect on tyramine decarboxylation. However, it was also found that at the same concentrations of NaCl and lactose, a higher amount of tyramine was detected under anaerobic conditions. In all strains tested, tyramine decarboxylation started during the active growth phase of the cells.