德氏乳杆菌保加利亚亚种耐酸机制

德氏乳杆菌保加利亚亚种是最具经济价值的乳酸菌之一,其耐酸能力是影响其发酵性能和发挥益生功效的关键因素。本文就该菌的耐酸能力调节进行阐述。     

Protein‐loaded sodium alginate and carboxymethyl cellulose beads for controlled release under simulated gastrointestinal conditions

Network characteristics of sodium alginate and carboxymethyl cellulose, in relation to the controlled release of bovine serum albumin under simulated gastrointestinal conditions, have been examined. Small‐deformation dynamic oscillation in‐shear, compression testing and optical microscopy were employed to monitor the structural characteristics of polysaccharide beads as excipients for protein entrapment. Work focused on gastrointestinal functionality and controlled release utilising variable acidity and counterion addition in preparations. Beads were found to shrink in gastric and swell in intestinal conditions, thus emphasising the disparate effect of experimental conditions. Changes in mechanical properties reflected alterations in gel microstructure, with beads becoming relatively strong in the gastric environment. In contrast, networks were weakened in the intestinal environment resulting in bead disintegration towards the end of digestion time. This type of structural behaviour allows the controlled delivery of the protein, as a model bioactive, in a particular part of the gastrointestinal tract.     

利用德氏乳杆菌全酶液水解牛乳酪蛋白

采用响应面法优化超声破碎工艺获取乳酸菌的全酶液,探讨超声破碎功率、超声破碎时间以及溶菌酶的添加量对超声破碎的影响,对获取的全酶液进行水解脱脂乳及酪蛋白的实验。结果表明：随着超声破碎功率的加大,超声破碎时间的增加,所得酶活力值均呈现先增长后下降的趋势,分析所得的回归方程确定最佳工艺修正参数为：超声破碎仪设定功率71%,超声破碎仪设定时间7.5 min,体系中溶菌酶的添加量290 μL/30 mL菌液。在此条件下获取的全酶液水解脱脂乳及酪蛋白显示,超声破碎的过程对酶活力有较大损失,分开处理其胞内和胞外酶液,水解脱脂乳和酪蛋白所获得游离氨基氮的含量分别提高了223.86%和324.11%,高效液相色谱分析结果也证明全酶液水解所得多肽含量及肽段大小多样性均有所提高。     

Morphology,FTIR fingerprint and survivability of encapsulated lactic bacteria (Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus) in simulated gastric juice and intestinal juice

Microencapsulation of bacteria in hydrocolloid beads is known as a potential way to enhance and protect their survivability in the digestive tract. We encapsulated a mix of two lactic bacteria (Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus) in sodium alginate (AG) and chitosan (CH) matrices. The morphological parameters of beads containing bacteria were comparatively measured. The FTIR fingerprint can discriminate the presence of bacteria into the beads, identifying specific absorption peaks for bacteria located at 1750, 2852 and 2926 cm^?1. To check the survivability of bacteria, beads were incubated in simulated gastric juice (pH 1.5) and intestinal juice (pH 7.2) for different periods of time, up to 120 min. The 2% AG beads, better than CH beads, provided best protection of bacterial survivability. Encapsulated mix of S. thermophilus and L. delbrueckii subsp. bulgaricus can behave as a probiotic bacteria, viable, surviving in the simulated gastric and intestinal juice.     

德氏乳杆菌保加利亚亚种自发突变抗噬菌体菌株的选育及其特性

利用德氏乳杆菌L.bulgaricus KLDS08021为宿主菌,采用次级感染的方法筛选自发突变的抗噬菌体菌株,将筛选出的有抗性的7株菌株进行RAPD-PCR鉴定,然后进行EOP、遗传稳定性、溶源性检测,并对其中的2株抗性菌BIM1、BIM7及野生株进行发酵性能测定。结果显示,7株菌RAPD-PCR电泳图谱一致,均来自宿主菌,是遗传性稳定的非溶源性菌;无论噬菌体是否存在2株抗性菌BIM1、BIM7其发酵活菌数、产酸能力都比野生株高,且制作的发酵乳黏度与硬度也比野生菌株强,适于作为酸奶发酵菌株。     

优良保加利亚乳杆菌菌株的筛选及增殖培养

以24株德氏乳杆菌保加利亚亚种菌株为材料,通过发酵特性分析,包括产酸、产粘、后酸化及发酵风味质地等感官特性,从中筛选出具有优良发酵特性的菌株L3,LB和M28.并对LB菌株的增殖培养基进行了初步探讨,为酸奶发酵剂的开发奠定一定基础.     

Detection and localization of a peptidoglycan hydrolase in Lactobacillus delbrueckii subsp. bulgaricus

Peptidoglycan hydrolase activities in Lactobacillus delbrueckii subsp. bulgaricus were detected by analysis of bacterial extracts on denaturing polyacrylamide gel electrophoresis containing lyophilized Micrococcus lysodeikticus cells as substrate. A hydrolase with an estimated molecular mass of 80 kDa was found to cross-react on Western blot with monoclonal antibodies raised against muramidase-2 of Enterococcus hirae. These antibodies were also used to demonstrate that the method of cell sample preparation affected protein detection. Slot and Western blots indicate that the peptidoglycan hydrolase from L. bulgaricus is bound to the cell wall. Immuno-labeling followed by optical and electron microscopic observations suggest that this hydrolase is intracellular and restricted mainly to the space between the membrane and the cell wall.     

Whey protein/alginate beads as carriers of a bioactive component

Bioactive components encapsulated in gel particles offer a means to deliver these components in a time release fashion to improve health through daily diet. The objective of this work was to evaluate the physical/mechanical properties of a release system consisting of whey protein/alginate gel beads in a model beverage. Riboflavin, vitamin B₂, was incorporated in the gel beads as a model bioactive compound. Gel beads were formed both with and without riboflavin and then added to a xanthan/sucrose suspending medium. Both the beads and the polymer solutions from which they were formed were characterized by standard rheological methods. An absorbance method was used to monitor riboflavin release over time. NMR and MRI (magnetic resonance imaging) methods characterized bead integrity and the state of protons in the gel beads. The combination of whey protein and alginate formed beads with good integrity and zero order rate kinetics for the release of riboflavin. The release rate of the bioactive component was more rapid that the typical shelf life of a bottled beverage however, the work represents a step toward the design of this type of functional food through characterization of the encapsulation medium.     

保加利亚乳杆菌LJJ自溶的影响因素分析

考察了不同生长阶段的保加利亚乳杆菌LJJ在不同温度及酸度环境中的自溶度变化,不同浓度SDS和Triton X-100对菌体自溶度和溶菌酶活性的影响,并采用扫描电镜观察菌体自溶中的形态变化,以确定LJJ自溶的最适条件并初步探讨溶菌酶在自溶中的作用。结果表明,处于对数生长期的LJJ菌体最易发生自溶。LJJ自溶度随环境温度(4⁵0℃)的升高,温育时间的延长而相应增大。LJJ自溶的最适温度为50℃,最适pH值范围为6.050℃)的升高,温育时间的延长而相应增大。LJJ自溶的最适温度为50℃,最适pH值范围为6.0⁸.0。0.50 g/L SDS对LJJ自溶和溶菌酶的抑制率分别为99.6%和97.4%。1.00 g/L Triton X-100分别将LJJ的自溶度和溶菌酶活性提升了20.7%和39.5%。扫描电镜观察表明,自溶细胞与正常细胞相比,细胞壁出现孔洞,细胞完整性遭到破坏而0.50 g/L SDS可抑制LJJ细胞的破坏。因此,环境pH、温度、温育时间、SDS浓度、Triton X-100浓度及菌体生长阶段可通过溶菌酶活性的作用显著影响LJJ的自溶,为调控乳酸菌自溶提供了初步理论基础。     

保加利亚乳杆菌生长性能与后酸化的关系初探

以5株德氏乳杆菌保加利亚亚种为出发菌株,详细研究了5株菌的生长代谢、酸敏感性及蛋白水解力,并分析了这些特性与后酸化之间的关系。结果表明:5株菌的生长性能与后酸化之间存在紧密联系。其中,菌株的生长速度和糖分解代谢能力与后酸化成正比;对酸较敏感菌株的后酸化也较弱;菌株的蛋白水解能力与生长速度和后酸化之间没有明显的相关性。     

冷冻和冻干过程对保加利亚乳杆菌存活性影响的研究

研究了预冻过程中冷冻温度、冷冻时间以及升华干燥过程中发酵剂含水量对保加利亚乳杆菌(Lactobacillus delbrueckii subsp.bulgaricus)L.b-S1细胞存活率的影响.结果表明:-20～-18℃和-40～-35℃冷冻,对细胞存活率无显著影响(P＞0.05);冷冻起始至完全冻结,细胞存活率差异显著(P＜0.05),而冻结后细胞存活率并不随冷冻时间延长而存在显著差异(P＞0.05);干燥过程中,发酵剂含水量对细胞干燥存活率影响显著(P＜0.05),但存活率降低程度因含水量下降水平不同而出现差异,其中以含水量由40%左右至5%左右的过程影响最大,细胞存活率由88.68%降至40.11%,下降约50%左右,差异显著(P＜0.05).本研究为研制开发高效浓缩型直投式酸奶冻干发酵剂,合理调控冻干工艺提供指导依据.     

德氏乳杆菌保加利亚亚种atpA抗原的表达及抗体制备

利用原核表达德氏乳杆菌保加利亚亚种atpA抗原并进行抗体制备,以便用于Western blot分析.首先利用PCR的方法扩增出atpA蛋白的基因片段,利用大肠杆菌pQE 40原核表达系统表达重组atpA抗原,镍柱亲和层析纯化表达蛋白后,常规免疫两只新西兰长耳白兔.利用ELISA检测兔血清的效价来制备多克隆抗体,然后用Protein A纯化抗体.最后得到的atpA抗体进行Western blot检测验证抗体的特异性.结果表明,两只新西兰长耳白兔的血清效价分别为1∶20000和1∶80000;Western blot检测表明,制备的抗体能够与H+-AT-Pase蛋白进行特异性的结合.本研究成功的在大肠杆菌中表达了德氏乳杆菌保加利亚亚种的atpA蛋白,并利用其制备了效价较高的多克隆抗体,该抗体可作为一抗用于Western blot分析.     

Isolation and characterisation of a Lactobacillus delbrueckii subsp. bulgaricus mutant with low H+‐ATPase activity

Lactobacillus delbrueckii subsp. bulgaricus is an acid‐tolerant species used in yoghurt production and considered important in postacidification reductions in palatability of yoghurt during storage. In this study, a variant (ND06‐2) of L. delbrueckii subsp. bulgaricus ND06 was selected using neomycin sulphate. ND06‐2 exhibited sensitivity to acids and lower H⁺‐ATPase activity compared with ND06. High‐pressure liquid chromatography (HPLC) was used to demonstrate that the capacity of ND06‐2 to produce lactic acid was significantly lower than for ND06 during fermentation or storage. These results suggest that the L. delbrueckii subsp. bulgaricus variant could be used in yoghurt starters and have a lower tendency to cause postacidification.     

嗜热链球菌和德氏乳杆菌保加利亚亚种共生关系的研究进展

嗜热链球菌和德氏乳杆菌保加利亚亚种是组成酸奶发酵剂的主要菌种,在长期共处于牛乳环境的过程中形成多种协同共生关系.这些协同共生关系赋予发酵剂菌株产酸快、黏度高、活性物质丰富等优良性能,同时对酸奶品质产生重要影响.该文旨在从营养供给、抵抗环境胁迫及酸奶品质改良等方面讨论两菌的协同共生关系,为提高发酵剂菌株活力及酸奶品质提供...     

保加利亚乳杆菌的筛选及其发酵乳中的风味物质

以分离自传统发酵乳中的7株保加利亚乳杆菌为试验菌株,测定发酵乳制作和贮藏(12 h)期间pH值、滴定酸度、活菌数、黏度的变化,并结合固相微萃取与气相色谱-质谱联用技术(SPME-GC-MS)检测贮藏12 h时发酵乳中的挥发性风味物质.结果表明,7株保加利亚乳杆菌中,IMAU62115在牛乳中的发酵时间最短(7 h),活...     

保加利亚乳杆菌水解酪蛋白能力及产物分析

为了深入研究保加利亚乳杆菌酪蛋白水解情况。本试验主要以保加利亚乳杆菌ATCC 11842作为研究对象,同时以保加利亚乳杆菌KLDS 08007、KLDS 08009、KLDS 08010、KLDS 08014和KLDS 08018作为对照研究,测定不同菌株发酵液的酪蛋白水解度和多肽含量,同时对ATCC 11842的酪蛋白水解产物进行SDS-PAGE电泳分析和LC-MS分析。结果表明:ATCC 11842的水解能力最强,水解度为13.89%,多肽含量为0.63 mg/m L。测定六株保加利亚乳杆菌细胞壁蛋白酶的活性与水解能力进行比较分析发现,该酶活性与菌株的水解能力呈正相关,ATCC 11842的酶活力最强,达到15.52 U/m L,比活力为6.73 U/mg。通过电泳图谱发现,随着发酵时间的延长,酪蛋白逐渐水解,并生成3.4~21.0 ku的多肽。对水解物进行LC-MS分析,酪蛋白水解后共产生77种6~25肽的片段,其中6~7肽占片段种类的9%,8~14肽占多肽种类的77%。     

保加利亚乳杆菌H+-ATPase弱化菌株的诱变选育

以保加利亚乳杆菌ND06(Lactobacillus delbrueckii subsp.bulgaricus ND06)为出发菌株,利用硫酸新霉素诱变、筛选出一株H+-ArPase 弱化菌株ND06-6.该菌株在MRS培养基中,37℃培养18 h时,H+-ATPase活性为0.65 μmol/(mg· min),和出发菌株相比,H+-ATPase活性下降了35％.在脱脂乳培养基中培养0,4,8,12,16,20,24,36,48和72 h后,诱变菌株ND06-6的乳酸、乳糖和半乳糖的的代谢能力明显低于出发菌株.这些研究结果为弱后酸化酸奶发酵剂的开发和应用提供了理论依据.     

Factors influencing autoaggregation and aggregation of Lactobacillus delbrueckii subsp. bulgaricus isolated from handmade yogurt

Of 26 Lactobacillus delbrueckii subsp. bulgaricus strains isolated from yogurt, strains B2 and 22, which produce low levels (28 and 21 mg liter(-1), respectively) of extracellular polysaccharides (EPSs), and strains B3 and G12, which produce high EPS levels (211 and 175 mg liter(-1), respectively), were selected for further study. The two high EPS-producing strains showed a significant autoaggregation and coaggregation ability with Escherichia coli ATCC 11230 (P < 0.05). Moreover, the effect of bile was evaluated on autoaggregation and hydrophobicity. Autoaggregation and hydrophobicity of these L. delbrueckii subsp. bulgaricus strains decreased after treatment with bile. Only the high EPS-producing L. delbrueckii subsp. bulgaricus strain B3 showed greater autoaggregation (80%) and hydrophobicity (86%) than the other strains after bile treatment. When these strains were assessed for the inhibition of E. coli ATCC 11230 in coculture, L. delbrueckii subsp. bulgaricus B3 completely inhibited E. coli during 24 and 48 h of incubation. This investigation showed that a high EPS production and coaggregation ability may be important in the selection of probiotic strains.     

具有优良发酵特性德氏乳杆菌保加利亚亚种的筛选及其益生特性

本研究对10株实验室保存的德氏乳杆菌保加利亚亚种产酸能力,产黏能力,产香能力和贮藏期稳定性进行测定,并通过测定乳酸菌对抗生素的耐药性、抑菌能力、β-半乳糖苷酶(β-Gal)活性、α-半乳糖苷酶(α-Gal)活性和乳酸脱氢酶(LDH)活性探究菌株的益生特性.单菌株发酵实验结果表明:菌株KSDB-1凝乳时间短(6.3 h)...     

Microbial and physiochemical changes in yoghurts containing different Lactobacillus delbrueckii subsp. bulgaricus strains in association with Lactobacillus plantarum as an adjunct culture

Two strains of slow and normal acid‐producing Lactobacillus delbrueckii subsp. bulgaricus, mixed separately with Streptococcus thermophilus, were used in the preparation of yoghurt with or without Lactobacillus plantarum A7. Viable cell counts of lactic bacteria, pH, lactic acid and acetic acid concentrations, soluble nitrogen content, spontaneous syneresis, viscosity and firmness were studied at selected points during the refrigerated storage period. Results showed that survival of L. plantarum A7 in yoghurt was independent of the acidification capability of L. delbrueckii subsp. bulgaricus strains which had already been characterised in their pure culture. Thus, in the preparation of probiotic yoghurt, using slow acid‐producing L. delbrueckii subsp. bulgaricus strain in combination with nondairy lactobacilli postfermentation acidification is not restricted but instability in other physicochemical properties of the yoghurt may be expected.