植物乳杆菌转化合成共轭亚油酸的培养基条件优化

生物法转化合成共轭亚油酸,产物中异构体组成单一,具有很好的应用前景。通过一株植物乳杆菌(Lactobacillus plantarum)合成共轭亚油酸的培养基成分进行优化,确定最优的培养基组成:葡萄糖20g/L,酵母浸出物40g/L,硫酸镁0.5g/L,硫酸锰0.5g/L,乙酸钠2g/L,磷酸氢二钾1g/L。优化后,共轭亚油酸产量达到0.259g/L,相比优化前(0.0455g/L)有了较大的提升。      

一株植物乳杆菌转化生成共轭亚油酸的特性研究

报道了一株植物乳杆菌(L.plantarumLT2-6)发酵转化亚油酸(LA)生成共轭亚油酸(CLA)的特性研究。微氧环境有利于CLA的生成;温度为37℃、初始pH为7.0、底物LA浓度为0.075%时,菌体生长及CLA生成量较高。时间曲线结果表明,接种后8h,CLA开始生成;发酵24h时,CLA生成量达到最高(0.29g/L),LA转化率为387%。生成的CLA产物主要为cis9,trans11/trans9,cis11-CLA。     

植物乳杆菌ZS2058转化亚油酸为共轭亚油酸条件的初步研究

研究了植物乳杆菌ZS2058转化亚油酸为共轭亚油酸的条件,在培养基中添加10g/L吐温-80制成LA-吐温-80胶束溶液,可以提高CLA的转化率,最佳转化温度为35℃,添加1mg/mL的LA有较高的转化率和CLA产量。在MRS、SKM、KPB三种转化体系中SKM的转化率较高。在含LA的培养基中预培养12h,可以提高CLA的转化率。最佳转化接种量在1%～2%之间。      

植物乳杆菌ZS2058生物转化共轭亚油酸反应条件的优化研究

对本实验室从泡菜中筛选到的植物乳杆菌ZS2058完整细胞生物转化共轭亚油酸的反应条件进行了系统研究。在1mL磷酸钾缓冲液反应体系中,通过单因素实验和响应面分析,确定最合适的反应条件为:亚油酸底物浓度为0.8mg/mL,细胞浓度为4×1010cfu/mL,反应温度为37℃,缓冲液pH为6.7。在此反应条件下,cis9,trans11-CLA的浓度为374.24μg/mL,转化率高达46.78%,这对于实现共轭亚油酸的高效生产和研究其生理功能具有重要的现实意义和理论价值。      

植物乳杆菌发酵豆乳对多囊卵巢综合征大鼠的影响

为探究植物乳杆菌发酵豆乳缓解大鼠多囊卵巢综合征症状(polycystic ovary syndrome, PCOS)的能力,采用来曲唑灌胃的方式构建PCOS大鼠模型,并以此模型评价了3株植物乳杆菌发酵豆乳对PCOS大鼠体重、卵巢病理学、性激素水平、糖脂代谢等的影响。结果表明,来曲唑给药后大鼠体重以及卵巢重量均显著增加,卵巢呈多囊样改变,性激素水平以及生化指标异常。3株植物乳杆菌发酵豆乳干预后均不同程度减轻了来曲唑引起的大鼠卵巢病变,降低PCOS大鼠血清睾酮、总胆固醇、甘油三酯及低密度脂蛋白胆固醇水平。其中,植物乳杆菌CCFM1019发酵豆乳可以显著增加血清中雌二醇水平,降低谷草转氨酶(aspartate aminotransferase, AST)含量,而植物乳杆菌LP4发酵豆乳可以显著降低血清中促黄体生成素含量以及谷丙转氨酶、AST含量。这些不同植物乳杆菌发酵的豆乳能在不同程度上缓解大鼠PCOS症状。     

嗜酸乳杆菌转化菜籽油生成共轭亚油酸条件的优化

文中对嗜酸乳杆菌静息细胞转化菜籽油生成CLA工艺进行了优化研究,通过单因素试验,确定了最适转化液介质、介质浓度、pH、温度、转化时间和细胞浓度,同时,研究了不同金属离子对亚油酸异构酶活性的影响。在该基础上,利用Box-Benhnken中心组合设计和响应面法对转化时间、pH、温度、细胞浓度进行了工艺优化分析,确定了在菜籽油浓度为7.5 mg/mL、脂肪酶的加入量为2 mg/30 mL时,以0.1 mol/L柠檬酸-柠檬酸钠缓冲液为转化液介质条件下,最优转化工艺条件是时间25 h,pH6.5,温度36℃,细胞浓度40 mg/mL,经优化后CLA的生成量可以达到230.12±7.52μg/mL。     

产共轭亚油酸植物乳杆菌的筛选、鉴定与诱变

从传统泡菜中筛选得到1 株乳酸菌lp15 能够合成共轭亚油酸(CLA),经16S rRNA 全序列分析法和API 系统鉴定法鉴定为植物乳杆菌(Lactobacillus plantarum)。利用人工诱变方法,以lp15 为出发菌株,采用紫外线、硫酸二乙酯(DES)依次诱变处理,经进一步液体发酵复筛获得多株突变菌株,CLA合成能力较出发菌株提高了29.3%～52.2%。其中lp15-2-1 突变菌株为CLA 生成能力最高菌株,MRS 培养液中添加0.2mg/mL LA 培养48h,CLA 产量达30.13μg/mL。经气相色谱(GC)检测分析,产物中cis9, trans11- 共轭亚油酸(c9, t11-CLA)占76.5%,trans10, cis12-共轭亚油酸(t10,c12-CLA)占23.5%。     

响应面法优化植物乳杆菌lp15-2-1产共轭亚油酸发酵条件

对植物乳杆菌(Lactobacillus plantarum)lp15-2-1 转化亚油酸生成共轭亚油酸(CLA)发酵工艺进行研究。通过单因素试验确定最佳接种量、培养温度、初始pH 值、培养时间、亚油酸添加时间、亚油酸质量浓度。采用响应面Box-Benhnken 试验设计,建立初始pH 值、培养温度和亚油酸质量浓度的二次多项式回归方程模型,所得植物乳杆菌发酵产共轭亚油酸的最佳参数为：温度30℃、亚油酸质量浓度0.21mg/mL、初始pH6.3,此条件下,CLA得率为44.77μg/mL,CLA 理论得率为45.326μg/mL,转化率为21.32%,与优化前转化率7.78% 相比,有了很大提高。     

植物乳杆菌亚油酸异构酶的分离纯化及其性质研究

经硫酸铵分级沉淀、阴离子交换层析和凝胶过滤,由植物乳杆菌(LactobacillusplantarumL 2 9)分离纯化得到亚油酸异构酶,分子量为4 3ku。对其酶学性质进行研究,结果表明,温度37℃、pH 6 0时酶活性较高;Co2 + 、Fe2 + 可提高酶的活性,Cu2 + 、Zn2 + 则对酶活力有抑制作用;该酶作用于亚油酸的Km=2 5 3×10 -5mol/L ,Vmax=2 5 7×10 -8mol/ (min·mg)。     

Metabolism of ferulic acid during growth of Lactobacillus plantarum and Lactobacillus collinoides

BACKGROUND: Food‐isolated lactic acid bacteria can transform ferulic acid (FA) into several products. Since quantification of these metabolites during the different bacterial growth phases is lacking, the aim of this study was to identify and quantify conversion products of FA and to follow the kinetics of FA metabolism during growth of Lactobacillus plantarum and Lactobacillus collinoides. RESULTS: Lactobacillus plantarum and Lactobacillus collinoides were incubated in MRS broth, to which different amounts of FA were added (final concentrations of 0, 0.5, 1.5 and 3 mmol L^?1), at 30 °C until the late stationary phase. Lactobacillus plantarum metabolised FA into 4‐vinylguaiacol (4‐VG) and hydroferulic acid (HFA). Conversion to 4‐VG started simultaneously with the degradation of FA, while formation of HFA started in the mid‐exponential phase. Lactobacillus collinoides only formed 4‐VG, mainly in the stationary phase. No significant effect of the different amounts of FA was seen on the growth and fermentation characteristics of both bacteria. CONCLUSION: The results demonstrate that both bacteria are able to convert FA. However, start of conversion differs between the two strains. The different amounts of FA had no influence on the growth and fermentation characteristics of both bacteria. Copyright © 2012 Society of Chemical Industry     

Response of a Lactobacillus plantarum human isolate to tannic acid challenge assessed by proteomic analyses

Scope : To gain insight on the mechanisms used by intestinal bacteria to adapt and resist the antimicrobial action of dietary tannins and identify targets for tannic acid in Lactobacillus plantarum. Methods and results : A proteomic analysis of an L. plantarum human isolate exposed to the tannic acid challenge was undertaken. Tannic acid targeted proteins involved in outstanding processes for bacterial stress resistance including cyclopropanation of membrane lipids, stress response at population scale and maintenance of cell shape. To respond to this aggression, tannic acid‐misfit cells of L. plantarum challenged with tannic acid reorganized their metabolic capacity to economize energy and express proteins involved in oxidative stress defense and cell wall biogenesis, indicating that the injury incurred by tannic acid was based on oxidative damage and disruption of the cell envelope. The induction of 3‐octaprenyl‐4‐hydroxybenzoate carboxy‐lyase, which is sensitive to changes in redox conditions and involved in ubiquinone biosynthesis in other bacteria, suggests for a tannic acid‐induced redox imbalance. Conclusion : The results reveal the adaptation of a gastrointestinal isolate of L. plantarum to tannic acid and identify antibacterial targets for this dietary compound. This provides the basis for the selection of tannin‐resistant microorganisms and their use to obtain health benefits from tannin‐containing diets.     

植物乳杆菌生物合成苯乳酸条件优化研究

以苯丙酮酸为底物,以植物乳杆菌静息细胞为催化剂,对苯乳酸生物合成条件进行了优化。 试验首先确定了不同因素对苯 乳酸产量的影响,然后通过正交试验优化了最佳合成条件。 结果表明,细胞培养时间为18 h时催化活性最高,且在温度为35 ℃,pH 为6.5,静息细胞、苯丙酮酸和葡萄糖质量浓度分别为80 g/L、3.0 g/L和4.0 g/L的催化条件下,经过4 h转化,苯乳酸产量达到最高,为 1.68 g/L。 在最优条件下,静息细胞重复利用4次,苯乳酸产量没有显著降低,表现出了植物乳杆菌转化苯丙酮酸合成苯乳酸的良好稳 定性。     

乙醇胁迫提高植物乳杆菌p-8的共轭亚油酸(CLA)转化率和相关酶转录水平

该文研究了在MRS培养基中添加0.05 mg/mL LA（Linoleic acid,LA）和不同浓度的乙醇时植物乳杆菌p-8的CLA（Conjugated linoleic acid,CLA）转化率和CLA合成相关酶转录水平的差异情况。结果显示,发酵液中的三种CLA异构体转化率都是在添加0.50%乙醇时最高,其中转化cis9,trans11-CLA（t9,t11-CLA）异构体最高,为2.49%,比不添加乙醇增加2.37倍。添加不同浓度乙醇的发酵液中trans10,cis12-CLA（t10,c12-CLA）转化率都是最低的。菌体中产生的CLA非常少,但规律与发酵液的基本一致。添加0.50%乙醇菌体中t9,t11-CLA转化率最高,其转化率仅为0.05%,比不添加乙醇增加了5倍。当乙醇浓度高于0.50%时,各种不同CLA异构体的转化率却都减少。结果表明CLA是在胞液内产生后再被运转到胞外的,一定浓度范围内的乙醇胁迫通过提高CLA合成相关的酶基因转录水平,进而促进了CLA的转化,可见CLA合成相关酶基因转录水平是造成CLA转化率差异的主要原因。结果为阐明植物乳杆菌p-8产CLA的分子机制和寻找有效提高CLA生成的调控手段奠定了基础。     

Production of free conjugated linoleic acid by Lactobacillus acidophilus and Lactobacillus casei of human intestinal origin

A gas chromatographic procedure was used for analysis of conjugated linoleic acid (CLA) isomers cis-9, trans-11-octadecadienoic; trans-10, cis-12 octadecadienoic; and trans-9, trans-11-octadecadienoic (c9t11, t10c12, t9t11) produced by lactobacilli. Four different cultures, two strains each of Lactobacillus acidophilus and Lactobacillus casei were tested for their ability to produce CLA from free linoleic acid in MRS broth supplemented with linoleic acid. Different concentrations of linoleic acid (0, 0.05, 0.1, 0.2 and 0.5 mg/ml) were added to MRS broth, inoculated with the lactobacilli, and incubated at 37 degrees C. Viable counts and amounts of individual isomers of CLA (c9t11, t10c12, t9t11) were measured at 0, 24, 48, and 72 h. All the cultures were able to produce free CLA in media supplemented with linoleic acid. Maximum production of CLA (80.14 to 131.63 microg/ml) was observed at 24 h of incubation in broth containing 0.02% of free linoleic acid. No significant (P > 0.05) increases in total CLA levels were observed after 24 h of incubation. The ability of the cultures to produce CLA in skim milk supplemented with 0.02% free linoleic acid also was studied. In this medium, the total amounts of free CLA after 24 h of incubation ranged from 54.31 to 116.53 microg/ml. The use of lactic acid bacteria able to form free CLA in cultured dairy products may have potential health or nutritional benefits. Free CLA in the products likely would be more readily available for absorption from the digestive tract than if it were incorporated into the cells of the starter culture.     

常压室温等离子体诱变选育高产酸植物乳杆菌

采用常压室温等离子体（ARTP）对从酸马奶中筛选的植物乳杆菌（Lactobacillus plantarum）YL15进行诱变,选育出产酸能力高并且发酵性能稳定的乳酸菌菌株,并对其遗传稳定性及最适生长温度和最适pH进行研究。结果表明,通过单因素试验确定最佳诱变条件为在120 W照射100 s,致死率为91.09%。通过溶钙圈法和酸度测定筛选出目标菌株YL15-4。初始菌株YL15产酸量为10×10-3 mol/L,诱变菌株YL15-4与初始菌株YL15相比较,其产酸能力提高了50%,产酸量达到15×10-3 mol/L,其最适生长温度和最适生长pH值分别37 ℃、5.0,并且遗传稳定性良好。     

一株强抑菌活性植物乳杆菌的分离及益生性能研究

以健康黔北黑猪肠道黏膜内容物为原料,采用厌氧罐培养法分离出肠道细菌,结合抑菌试验筛选出其中具有强抑菌活性菌种进行分子生物学鉴定,并对其耐受性及抑菌物质的稳定性进行研究。结果表明,该菌株被鉴定为植物乳杆菌（Lactobacillus plantarum）,对金黄色葡萄球菌ATCC6538、大肠埃希氏菌ATCC25922和鼠伤寒沙门氏菌ATCC14028等指示菌具有强抑制作用,其发酵上清液对3种指示菌的最大抑菌圈直径分别约为44.7 mm、23.1 mm和31.8 mm;菌株发酵上清液抑菌物质耐高温,120 ℃处理30 min抑菌活性无显著变化（P＞0.05）;胰蛋白酶处理后,上清液的抑菌圈直径显著减小（P＜0.05）,而过氧化氢酶和胃蛋白酶处理后抑菌圈直径无显著变化（P＞0.05）,表明主要抑菌物质为有机酸和活性肽;菌株经pH 2、0.3%猪胆盐或人工胃肠液处理6 h后存活率分别为0.06%、0.61%、40.80%和25.65%,显示出良好的益生性能。     

Improved sauerkraut production with probiotic strain Lactobacillus plantarum L4 and Leuconostoc mesenteroides LMG 7954

Probiotic strain Lactobacillus plantarum L4 and strain Leuconostoc mesenteroides LMG 7954 were applied for the controlled fermentation of cabbage heads. The parameters of the controlled and spontaneous fermentations, including antimicrobial effect of cabbage brines obtained at the end of both fermentations, were monitored. To check out the influence of starter culture strains, 10 randomly chosen lactic acid bacteria, isolated at the end of controlled cabbage heads fermentation were identified by API 50 CHL test, and the presence of the probiotic culture was confirmed by Pulsed Field Gel Electrophoresis. The starter cultures applied for cabbage heads fermentation allowed lowering of NaCl concentrations from 4.0% to 2.5% (w/v), considerably accelerated fermentation process by 14 d, and improved the product quality. The produced sauerkraut heads are considered probiotic product as viable probiotic cells count in final product was higher than 10(6) colony-forming units (CFU) per gram of product. PRACTICAL APPLICATION: The results of this research could be applied in the production of fermented cabbage heads with added functional (probiotic) value and with lower NaCl concentration with expected shortened fermentation time. This could not only be of economic but also of ecological importance.     

共轭亚油酸高产菌株的分离与鉴定

共轭亚油酸(CLA)因具有抗肿瘤、增强免疫力、抗动脉粥样硬化、减肥、抗氧化等功效,在医药与保健食品领域显示了巨大的应用潜力。目前获得CLA的主要途径是化学合成法,化学合成法产量高,但产物异构体众多,成分复杂,分离提纯困难,生物法因其反应条件温和,产物成分较单一,在食品医药领域展现出了良好前景。为筛选共轭亚油酸高产菌株,本文在有氧条件下,采用MRS培养基从泡菜环境中分离出16株产共轭亚油酸的菌株,其中PC-3菌株具有较高的CLA转化率,且能将亚油酸转化为具有生物活性的c9,t11-CLA和t10,c12-CLA。经形态学特征(扫描电子显微镜)分析、生理生化试验分析(API-50CHL)及16S r DNA序列分析及同源性分析,PC-3菌株被鉴定为植物乳杆菌(Lactobacillus plantarum PC-3),其16S r DNA基因序列长度为1460 bp,该序列Gen Bank的登陆号为JX270774.1。