Acid Production of Crude Fermentation in vitro Lactic Maize Resistant Starch in the Simulated and the Impacts of Some Exogenous Acid Bacteria Added

在模拟大肠环境（厌氧和37℃）下,利用健康成人粪便提取物发酵粗玉米抗性淀粉,研究短链脂肪酸的产生情况,并剖析外源乳酸菌对产酸模式的影响。实验结果表明,粪便提取物单独发酵抗性淀粉时,短链脂肪酸含量随发酵时间增加而呈增加趋势;粪便提取物和2株乳酸杆菌共同发酵粗玉米抗性淀粉时,发酵产物中乙酸含量增加,丙酸和丁酸含量没有明显变化;当粪便提取物和2株乳酸球菌共同发酵粗玉米抗性淀粉时,发酵产物中丙酸和丁酸含量明显增加,乙酸含量没有明显变化。研究结果表明,外源性乳酸菌影响粗玉米抗性淀粉的模拟肠道发酵产酸模式。     

外源乳酸菌对低聚异麦芽糖肠道发酵产酸的影响

利用健康成人粪便提取物体外模拟低聚异麦芽糖的肠道发酵,采用气相色谱法定量分析3种短链脂肪酸(乙酸、丙酸和丁酸)的生成量,揭示6株外源乳酸菌对低聚异麦芽糖肠道发酵产酸模式的影响。结果表明:粪便提取物单独发酵低聚异麦芽糖12~48 h时可产生短链脂肪酸,且其含量随发酵时间的延长而增加。分别添加6株外源乳酸菌于发酵体系后,发酵产物中3种短链脂肪酸的生成量明显增加,其中丁酸量增加最为显著,从0.12 mmol/L增至6.55~12.45 mmol/L。不同菌株促进低聚异麦芽糖肠道发酵产酸能力不同,从整体上看,屎肠球菌和鼠李糖乳杆菌促进低聚异麦芽糖产生3种短链脂肪酸的能力最强。     

抗性淀粉的体外产酸发酵研究

分别采用健康成人和健康婴儿的新鲜粪便提取液,在模拟大肠环境下(厌氧,37℃),作用于不同含量的抗性淀粉,发酵不同时间,采用气相色谱法测定不同发酵时间的发酵产物中短链脂肪酸的含量。分析结果表明,对于2种不同的粪便提取液,随着发酵时间的延长和抗性淀粉含量的增加,发酵产物中短链脂肪酸的总含量均逐渐增加,特别是丁酸含量增加显著;但在婴儿粪便提取液的发酵产物中,短链脂肪酸的含量,尤其是丁酸的含量,显著高于成人粪便提取液的发酵产物。     

外源肠道菌和玉米抗性淀粉对小鼠结肠菌群发酵产酸的影响

在37℃、厌氧条件下,通过向营养肉汤培养基中添加小鼠结肠内容物悬液模拟结肠环境。通过在模拟结肠环境中添加玉米抗性淀粉、单一及混合外源肠道菌及其组合,研究玉米抗性淀粉体外发酵产酸的特性及外源肠道菌对其的影响。结果表明:相对于未添加抗性淀粉或仅添加单一外源菌的组,添加3%(相对于营养肉汤培养基干基)玉米抗性淀粉后,乙酸、丙酸和丁酸的含量在发酵过程中(6、12、18、24、30 h)能够极显著(P0.01)地提升。在含有抗性淀粉的培养基中添加单一外源菌,嗜酸乳杆菌能够促进发酵产乙酸,青春双歧杆菌和粪肠球菌能够促进发酵产丙酸和丁酸,而大肠埃希氏菌会抑制产丙酸和丁酸。在含有抗性淀粉的培养基中添加混合外源菌,嗜酸乳杆菌和青春双歧杆菌能够协同发酵产乙酸、丙酸和丁酸,粪肠球菌与青春双歧杆菌组合发酵时,能够提升丙酸和丁酸产量,而大肠埃希氏菌则会不同程度地抑制其他3种菌发酵抗性淀粉产乙酸、丙酸和丁酸。     

不同人体肠道菌群利用厚黑拟层孔菌胞外多糖产酸特性的研究

利用真菌培养基及冻干技术提取液体发酵条件下厚黑拟层孔菌的胞外多糖,利用人体(成人及儿童)粪便提取物作为发酵底物,在模拟人体肠道(厌氧及37℃)环境条件下进行系统发酵,对发酵后体系的短链脂肪酸(甲酸、乙酸、丙酸及丁酸)进行检测。研究结果表明:以成人及儿童粪便提取液作为发酵底物模拟人体肠道环境下,短链脂肪酸的含量随发酵时间的延长而增加,同时也随着胞外多糖浓度的增加而有显著增加。儿童粪便发酵体系中产生的短链脂肪酸要高于成人粪便发酵条件下产生的短链脂肪酸。     

小米和燕麦中水溶性膳食纤维结构表征及对体外发酵体系短链脂肪酸的影响

利用酶解的方法提取黄小米和燕麦中水溶性膳食纤维,并利用红外光谱和高效液相色谱技术分别对其官能团结构及单糖组成进行表征;通过模拟人体肠道消化系统环境,对添加不同浓度的膳食纤维发酵体系进行短链脂肪酸含量测定,观察不同浓度及不同发酵时间对体外发酵液中短链脂肪酸的影响。研究表明黄小米和燕麦中水溶性膳食纤维具有多糖特征官能团,组成黄小米和燕麦中水溶性膳食纤维的单糖为鼠李糖、阿拉伯糖、葡萄糖和半乳糖;人体粪便提取物中的微生物可以代谢黄小米和燕麦中水溶性膳食纤维产生甲酸、乙酸、丙酸和丁酸,当向发酵体系中添加水溶性膳食纤维时,随着发酵时间的延长,短链脂肪酸的含量逐渐增加;同时随着添加膳食纤维的浓度的增加,短链脂肪酸的含量也逐渐增加。儿童粪便提取物发酵体系产生的短链脂肪酸浓度要高于成人粪便提取物发酵体系产生的短链脂肪酸浓度。     

双歧杆菌对香蕉抗性淀粉的体外发酵研究

通过体外发酵模拟,研究香蕉抗性淀粉对肠道微生物双歧杆菌的增殖作用,以及双歧杆菌对香蕉抗性淀粉的发酵作用。香蕉抗性淀粉经过人工胃液和人工肠液模拟体外消化后,以消化产物为底物,接种大鼠粪便中经特异性培养的双歧杆菌,在厌氧条件下进行体外发酵,分别在发酵4、8、12、16、20 h后取样,测定发酵液的p H值、短链脂肪酸和乳酸的含量。结果表明:培养基中添加香蕉抗性淀粉后,双歧杆菌的数量增加了3.51 log cfu/m L。随着发酵时间的延长,p H值逐渐降低,短链脂肪酸和乳酸含量逐渐升高。发酵20 h后,短链脂肪酸总酸含量达到118.52 mmol/L,乳酸含量达到8.147 mmol/L。实验证明:香蕉抗性淀粉有利于肠道双歧杆菌的生长,并能够被肠道菌利用,维持肠道的酸性环境。     

清香型白酒不同生产期的产酸环境差异性研究

为探究清香型白酒立醅与挑醅两个发酵期的产酸环境差异,对两个时期发酵过程中不同关键时间节点的酒醅酸度、产酸菌生物量、短链脂肪酸及对应酒体的短链脂肪酸进行定量分析。研究结果表明,两个时期酸度均随发酵进程递增,挑醅期在整个发酵过程酒醅酸度高于立醅期,且递增速度更快,两个时期在出缸时酸度相差幅度最大为2.22。立醅期发酵过程乳酸菌总数呈马鞍形变化,存在二次发酵现象;挑醅期发酵过程乳酸菌总数变化呈先增后减趋势,产酸菌生物量高于立醅期。在短链脂肪酸差异上,整体来看,挑醅期发酵过程的短链脂肪酸含量均高于立醅各指标含量,且己酸、戊酸及丙酸在挑醅发酵起始含量就较高,其在整个挑醅发酵过程中高表达,尤其是丙酸,两时期发酵末期差异倍数最高,且立醅期产酸最高水平只接近于挑醅期7对(天,下同)时。同样的,对两个时期酒体短链脂肪酸进行对比发现,除己酸外,热季挑醅期的短链脂肪酸含量均高于立醅期各指标含量。显然,挑醅发酵环境对产酸水平有很大影响,尤其是除乙酸外的其他短链脂肪酸的变化。     

苦荞抗性淀粉体外益生元的作用

为建立体外模拟发酵模型,以苦荞抗性淀粉为研究对象,分别对苦荞抗性淀粉作用下的小鼠粪便发酵液中p H、肠道主要有益菌(双歧杆菌、乳酸杆菌)、主要有害菌(大肠杆菌、产气荚膜菌)以及短链脂肪酸(乙酸、丙酸、丁酸,乳酸)的含量进行测定,研究了苦荞抗性淀粉对肠道益生特性的作用。试验结果表明:与对照组相比,添加不同浓度的苦荞抗性淀粉均可有效降低有害菌数量,其中高剂量组分别使大肠杆菌、产气荚膜菌减少了11.94%,21.70%(32 h),且不同剂量组的苦荞抗性淀粉使小鼠粪便发酵液pH分别下降49%, 50%和51%(32 h);此外,高剂量组较对照组分别使乙酸、丙酸、丁酸、乳酸的浓度增加了6.33, 66.50, 29.25和29.24倍(32 h)。同时,苦荞抗性淀粉组的益生元指数较对照组分别提高了22.42%, 29.50%和38.64%(32 h)。综上所述,苦荞抗性淀粉提高肠道中短链脂肪酸含量,从而降低肠道环境中p H。说明苦荞抗性淀粉可以促进有益菌的增长,改善肠道菌群结构。综上表明:苦荞抗性淀粉可作为一种益生元,在肠道中发挥益生作用。     

5 种食品原料对高脂饮食诱导的肥胖大鼠肠道菌群和短链脂肪酸的影响

香蕉粉、魔芋精粉、抗性糊精、玉米淀粉以及左旋肉碱这5 种食品原料据报道都有减肥功能,但它们对肠道微生物和代谢产物的影响以及与肠道菌群相关的减肥机制的差异尚不清楚。本研究采用16S rRNA测序检测肥胖大鼠肠道内容物,并通过高效液相色谱法（high performance liquid chromatography,HPLC）测定粪便中短链脂肪酸的含量。结果显示：益生菌属Ruminococcus_2、Coprococcus_2和Ruminiclostridium_5分别在玉米淀粉、香蕉粉和抗性糊精组中富集。Spearman相关性分析和典型关联分析（canonical correlation analysis,CCA）结果表明,肠道微生物的变化与糖脂代谢相关的生化参数密切相关。以可溶性膳食纤维为主的原料（魔芋粉和抗性糊精）和以不可溶性膳食纤维（抗性淀粉）为主的原料（香蕉粉和玉米淀粉）对短链脂肪酸的产生能力贡献度相当,均优于左旋肉碱。5 种原料增殖肠道益生菌、抑制有害菌的效果依次为抗性糊精＞香蕉粉＞玉米淀粉＞魔芋精粉,增加短链脂肪酸含量的能力依次为香蕉粉＞抗性糊精＞玉米淀粉＞魔芋精粉,而左旋肉碱对肠道菌群和短链脂肪酸几乎没有什么影响。CCA结果显示,糖脂代谢生化指标与肠道微生物群落的相关程度为空腹血糖水平＞高密度脂蛋白胆固醇水平＞总胆固醇水平＞低密度脂蛋白胆固醇水平＞总甘油三酯水平。Spearman相关性分析表明Ruminococcus_2与丁酸含量呈正相关,Escherichia.Shigella与乙酸、丁酸和总短链脂肪酸含量呈正相关。     

Structural Characteristics of Cellulose and Xylan during in vitro Fermentation by Pig Fecal Bacteria

In this study,cellulose and xylan were in vitro fermented by pig fecal bacteria.Rapid fermentation (40 h) and extended fermentation (eight weeks)were performed.The properties and ultra-structure changes of post-fermented solid residues were studied.In the end effluent,acetic acid,propionic acid,and butyric acid were observed to be the principal short-chain fatty acids (SCFAs) produced by anaerobic fermentation.Xylan was more accessible to bacteria than cellulose,leading to higher SCFA and lactic acid production.In addition,the crystalline structure of cellulose changed,leading to 16.3％ and 42.1％ increases in crystallinity index for rapid and extended fermentation,respectively.Through this research,a systematic and advanced method to study the degradation chemistry of cellulose and xylan during fermentation was developed.     

Resistant starch prepared from high-amylose maize starch with citric acid hydrolysis and its simulated fermentation in vitro

Resistant starch (RS) was prepared from high-amylose maize starch through two autoclaving–cooling cycles and then acid hydrolysis of retrograded starch. Experimental results showed that hydrolysis of retrograded high-amylose maize starch with 0.1 mol L⁻¹ citric acid at room temperature for 12 h would increase RS yield to 39%. At simulated conditions of large intestine (anaerobic and 37 °C), the prepared RS product was fermented in culture by fresh feces extract from healthy adult or healthy infant to produce short chain fatty acids. Formic, acetic, propionic and butyric acid produced in culture were analyzed by GC with capillary column. The GC analysis results showed that as the increase of fermentation time and the addition level of RS in culture, the production of short chain fatty acids was increased. However, the production of short chain fatty acids (especially butyric acid) in culture fermented by healthy infant feces extract was much higher than that fermented by healthy adult feces extract. It suggested that the production of short chain fatty acids from RS in simulated intestinal conditions might be affected by the intestinal microflora.     

中性大蒜果聚糖体外发酵产短链脂肪酸

大蒜果聚糖是大蒜深加工后废弃蒜渣中的主要成分,不能被人体消化利用,可选择性增殖肠道益生菌,具有开发成新型益生元的潜力。体外发酵产短链脂肪酸的分析对进一步评价大蒜果聚糖对肠道微生态的影响有重要意义。方法:本研究通过体外模拟人体肠道系统的pH、温度和营养条件,接种人体新鲜粪便悬浮液进行间歇式静态厌氧培养,采用高效液相色谱法(HPLC)分析不同聚合度(平均DP为11、16、21)的中性大蒜果聚糖为唯一碳源,培养0、4、8、12、24 h产短链脂肪酸的情况。结果:发酵12 h时短链脂肪酸浓度达到最高,之后便开始下降。其中样a(平均DP为11)的发酵液中总SCFA浓度最高,达86.38μmol/mL。总短链脂肪酸产量为样a(DP 11)>样b(DP16)>样c(DP 21)。24 h时,各发酵液的短链脂肪酸中丁酸和乳酸的比例较乙酸和丙酸高。结论:大蒜果聚糖能被人体肠道微生物发酵利用在体外产短链脂肪酸。其中丁酸和乳酸的摩尔浓度较乙酸和丙酸高。聚合度低的大蒜果聚糖产酸高于聚合度高的果聚糖。     

Effects of phosphorylated cross-linked resistant corn starch on the intestinal microflora and short chain fatty acid formation during in vitro human fecal batch culture

Changes in the intestinal microflora and formation of short chain fatty acids (SCFAs) were studied in an in vitro human fecal batch culture in a medium containing 10mg/mL of phosphorylated cross-linked resistant corn starch (RS4 type) as the sole carbon source. Viable counts of total anaerobic bacteria and bifidobacteria were higher for RS4 than for corn starch, RS2 (Novelose 240), and cellulose. However, viable counts of lactobacilli were decreased for RS4. The total short chain fatty acids (SCFAs) amount was also increased for RS4, corn starch, and RS2, but not for cellulose. The molar ratios of acetic:propionic:butyric acid were 39:20:15 after 36 h of culture in RS4 corn starch. RS4 corn starch promoted the growth of total anaerobic bacteria and bifidobacteria together with SCFAs formations in an in vitro human fecal batch culture.     

基于体外模型研究广叶绣球菌多糖的酵解特性

目的:通过人体新鲜粪便酵解模型研究广叶绣球菌多糖在体外的酵解特性,以便为其体内的消化特性研究提供理论支持.方法:制备广叶绣球菌多糖和人体粪便菌群混合液,在无氧环境下发酵0、1、2、6、12、24、48h后,检测酵解液中的OD600值、pH值,总糖、还原糖、短链脂肪酸(乙酸、丙酸、丁酸)含量,乳酸杆菌、双歧杆菌和大肠杆菌...     

Characterization of potentially probiotic lactic acid bacteria isolated from olives: Evaluation of short chain fatty acids production and analysis of the extracellular proteome

Probiotic strains can exert positive effects on human health by various mechanisms, among which the production of short chain fatty acids (SCFA). All the SCFA, mainly acetic, propionic and butyric acid, display beneficial effects on human health; butyric acid is the most interesting for its role in the prevention and treatment of colonic diseases.In this study the ability of 17 potentially probiotic food-isolated lactic acid bacteria to produce SCFA, directly or indirectly through the production of lactic acid, was investigated. Propionic and butyric acids were quantified by gas chromatography; acetic and lactic acids were quantified by specific enzymatic kits. All the tested strains displayed the ability to produce significant amounts of acetic and lactic acids (in the range of g/L) and just small amounts of propionic and butyric acids (in the range of mg/L).The extracellular proteomes of two of these strains, Lactobacillus plantarum S11T3E and Lactobacillus pentosus S3T60C, were evaluated by coupling 2-DE and MALDI TOF-TOF mass spectrometry. This is an interesting approach to investigate a probiotic strain, since secreted proteins represent the first contact point between bacteria and the host after ingestion. Six and seven proteins, in different isoforms, were identified from L. pentosus S3T60C and L. plantarum S11T3E, respectively. All of them have a predicted extracellular location, indicating the effectiveness of the used protocol. L. plantarum S11T3E secretes several proteins with adhesive function, suggesting that in this strain the ability to adhere to gut mucosa depends on this kind of molecules. In L. pentosus S3T60C just one adhesive protein is secreted suggesting that other families of molecules play a role in its adhesive ability.     

衡水老白干酒中产酸微生物的分离鉴定及性能研究

有机酸是白酒的重要呈味物质,也是香味成分的前驱物质。利用气相色谱和高效液相色谱检测出衡水老白干酒中含有乙酸、丙酸、异丁酸、丁酸、异戊酸和乳酸等6种有机酸。经试验在大曲、酒醅中分离得到69株产酸菌,其中具有代表性的醋酸菌1株,乳酸菌4株,丙酸菌2株。对其中一株主要产酸菌进行了16S rDNA鉴定为食窦魏斯氏菌（Weissella cibaria）,并对其的产酸特性进行初步研究,结果表明,在初始pH值6.5,接种量7%,温度32 ℃,培养36 h时乳酸产量最高,为7.12 mg/mL。     

凝固型苦荞酸奶对小鼠肠道菌群的调节作用

通过动物实验,利用实时荧光定量聚合酶链式反应方法、气相色谱-质谱、Illumina高通量测序技术,对小鼠肠道菌群数量和种类,以及主要代谢产物——短链脂肪酸（乙酸、丙酸、丁酸、乳酸）的含量进行分析,探究凝固型苦荞酸奶对小鼠肠道菌群的调节作用。结果表明：与对照组相比,凝固型苦荞酸奶使肠道中3 种有益菌（乳酸杆菌、双歧杆菌、柔嫩梭菌）的数量分别增加了523.81、911.01、12.18 倍,3 种有害菌（肠杆菌、肠球菌、拟杆菌）的数量分别减少了99.89%、97.31%、99.02%。同时,凝固型苦荞酸奶组小鼠粪便中的乙酸、丙酸、丁酸、乳酸的含量分别增加了36.87%、75.59%、56.26%、92.36%,且效果优于普通酸奶组。高通量测序结果表明,凝固型苦荞酸奶较普通酸奶可以显著增加小鼠肠道菌群多样性,在门水平上显著降低厚壁菌门和疣微菌门的丰度,显著提高拟杆菌门的丰度（P＜0.05）;且在属水平上显著提高norank_f__Bacteroidales_S24-7_group、Faecalibaculum菌属的相对丰度,显著降低Ruminococcaceae_UCG-014、unclassified_f__Lachnospiraceae、Lachnospiraceae_UCG-006菌属的相对丰度（P＜0.05）,提高肠道菌群优势菌属的地位。综上所述,凝固型苦荞酸奶相较于普通酸奶,可以更好地促进小鼠肠道内有益菌的增殖并抑制有害菌的增殖,促进短链脂肪酸的生成,增加肠道菌群多样性,优化菌群结构。     

Butyric Acid Spoilage of Fermented Cucumbers

Small cucumbers brined to equalize at 2.3% NaCl in an anaerobic tank underwent a normal primary lactic acid fermentation that resulted in 1.09% titratable acidity (as lactic) and pH 3.7. Nine months later the product was observed to have spoiled, as evidenced by an unpleasant odor. Products formed during spoilage in order of concentration were acetic acid > butyric acid > n-propanol > propionic acid. No lactic acid remained. No botulinal toxin was detected. Clostridium tertium was identified as contributing to the spoilage, but did not produce propionic acid or n-propanol under test conditions. Evidence indicated that unidentified bacteria, possibly propionibacteria sp., degraded lactic acid causing a rise in pH which allowed C. tertium to grow.