抑制黑色素合成的乳酸菌胞外多糖的筛选和性质研究

【目的】筛选可抑制黑色素合成的乳酸菌胞外多糖。【方法】通过观察凝乳拉丝外观筛选产胞外多糖的乳酸菌菌株,测量胞外多糖对B16黑色素瘤细胞黑色素合成和细胞活力的影响。对胞外多糖进行纯化,并通过PMP衍生-HPLC、红外光谱、抑制酪氨酸酶活性、抗氧化能力对其单糖组成和结构、作用机制进行研究。【结果】筛选到一株乳酸菌Lactobacillus rhamnosus HLAB122,发酵产生的胞外多糖在5 g/L浓度下可使B16细胞黑色素产量下降至空白对照的32.7%,且在96 h内对细胞活力无影响。纯化后的多糖由鼠李糖、葡萄糖、半乳糖构成,各单糖摩尔比为1?5.44?5.37。该胞外多糖不抑制酪氨酸酶活力且抗氧化性微弱。【结论】L.rhamnosus HLAB122产生的胞外多糖在个人护理产品中有潜在应用价值。     

一株乳酸菌胞外多糖产生的影响因素及其提取

应用苯酚一硫酸法对乳酸菌胞外多糖产生的影响因素进行了研究,表明该菌株在培养温度为30℃,培养时间为4048h,pH值降到4时,胞外多糖的产量最大。葡萄糖是乳酸菌产生多糖的良好碳源。在对乳酸菌的培养物进行离心、透析、脱蛋白、脱色,最后用乙醇沉淀,得到粗品多糖,粗品多糖至少含有两种分子量和含量相差很大的多糖。经过SephadexG-200凝胶柱得到多糖精品EPS—Ⅱ,薄层层析结果显示其为一纯化的样品。     

乳酸菌胞外多糖的研究进展

乳酸菌胞外多糖是乳酸菌生长代谢过程中产生并分泌到细胞外的一种天然高分子聚合物。作为一种新型的天然食品添加剂,乳酸菌胞外多糖因其独特的生理功能和产业潜力而备受研究者青睐。但由于乳酸菌胞外多糖结构组分、功能效用的不同,很难建立一个通用的生产方法和检测标准。另外,如何提高胞外多糖产量也是未来要面临的一大挑战。从乳酸菌胞外多糖的遗传研究、结构修饰、构效关系、生物学活性几个方面进行综述,并对未来研究方向进行展望。     

不产生胞外多糖的假单胞菌突起菌株E16

在适宜培养条件下,Ｐｓｅｕｄｏｍｏｎａｓ ｓｐ ３１２６０能将木糖转化为酸性胞外多糖（ＥＰＳ）,用甲基磺酸乙酯（ＥＭＳ）诱变处理Ｐｓｅｕｄｏｍｏｎａｓ ｓｐ ３１２６０得到一株完全不产生胞外多糖的突变菌株Ｅ１６。     

乳酸菌胞外多糖对人类肠道菌群的影响

摘要：胞外多糖（EPS）是微生物在生长代谢过程中分泌到细胞壁外及周围生长介质中的一类生物大分子物质。乳酸菌胞外多糖除了具有重要的加工优势外,如提高酸奶等乳制品的粘度、质地和口感,还具有重要的生理活性。本文重点介绍了乳酸菌胞外多糖的分类和主要生物活性,并从胞外多糖的益生元特性、作为肠道能源物质、对肠道细胞的粘附、调节肠道菌群多样性、调节肠道免疫五个方面综述了乳酸菌胞外多糖对人类肠道菌群的影响,同时对胞外多糖的后续研究提出了自己的建议。     

乳酸菌胞外多糖对人类肠道菌群的影响

胞外多糖(EPS)是微生物在生长代谢过程中分泌到细胞壁外及周围生长介质中的一类生物大分子物质。乳酸菌胞外多糖除了具有重要的加工优势外,如提高酸奶等乳制品的粘度、质地和口感,还具有重要的生理活性。本文重点介绍了乳酸菌胞外多糖的分类和主要生物活性,并从胞外多糖的益生元特性、作为肠道能源物质、对肠道细胞的粘附、调节肠道菌群多样性、调节肠道免疫五个方面综述了乳酸菌胞外多糖对人类肠道菌群的影响,同时对胞外多糖的后续研究提出了自己的建议。     

芽孢杆菌属产生的胞外多糖的研究

不同芽孢杆菌能产生不同的胞外多糖,根据多糖中的中性糖组成,可以把产生多糖的芽孢杆菌划分为三大类：第一类为中性糖由甘露糖和葡萄糖组成的菌株,计有B. subtils, B. pu-milus, B alvei B. cereus, B. sphaerieus, B. lichniformis, B. laterosporus, B．Pulvifaciens,B.brevis,B．Mycoides,B．Polymyxa,B．Popilliae,B．Macerans,B. fitmus 和 B. megaterium; 第二类为中性糖由甘露糖,葡萄糖和半乳糖组成的菌株,计有 B.polymyxa,B. larvae, B. thermop-hilus 和 B．Mycoides;第三类为中性糖由四种以上单糖组成的菌株,计有B. megatetium, B.coagulans 和 B．Circulans B．Polymyxa 和 B．Larvae产生的胞外多糖是一种类琼脂型的多糖,它们的水溶液(1％)具有加热熔化,冷却后凝固的特征,这种多糖是酸性多糖,它由甘露糖、葡萄糖,半乳糖和糖醛酸组成。     

一株乳酸菌胞外多糖产生的影响因素及其提取*

应用苯酚—硫酸法对乳酸菌胞外多糖产生的影响因素进行了研究,表明该菌株在培养温度为30℃,培养时间为40~48h,pH值降到4时,胞外多糖的产量最大。葡萄糖是乳酸菌产生多糖的良好碳源。在对乳酸菌的培养物进行离心、透析、脱蛋白、脱色,最后用乙醇沉淀,得到粗品多糖,粗品多糖至少含有两种分子量和含量相差很大的多糖。经过SephadexG-200凝胶柱得到多糖精品EPS-Ⅱ,薄层层析结果显示其为一纯化的样品。     

胞外多糖产生菌的筛选鉴定及其促生改土作用

【背景】一些微生物具有较强的产胞外多糖潜力,在改良盐碱土壤、促进作物生长方面潜力巨大。【目的】筛选耐盐碱且产胞外多糖的微生物,为开发具有盐碱土壤改良效果的菌剂提供菌种资源。【方法】从滨海盐碱植物根际土中筛选可在盐碱土壤中增殖、产胞外多糖、解有机磷和钾等能力的微生物菌株,并通过形态、生理生化、基于16S rRNA基因序列和gyrB基因的分子鉴定确定菌株类别,分析该菌株的生长、产胞外多糖特性受盐碱胁迫的影响,考察该菌对盐碱土壤不同粒径团聚体比例的影响,并结合盐碱土壤番茄种植试验考察对植物生长指标和土壤指标的影响。【结果】从727株滨海盐碱土壤菌株中筛选出一株产胞外多糖、具有较强盐碱土壤增殖能力且能解有机磷和钾的芽孢杆菌GBW HF-98,确定该菌为枯草芽孢杆菌沙漠亚种(Bacillus subtilis subsp. inaquosorum)。该菌可耐受pH值达10.0和NaCl浓度达110 g/L,在改善轻和中盐度土壤中不同粒径团聚体比例的效果较为显著。该菌的高剂量浓度T2组,在中度盐碱土壤的番茄种植试验中,与对照相比显著增加了壮苗指数、根干重、全株干重、茎粗和株高等指标,增幅分别为33.8%、59.3%、37.2%、12.3%和15.6% (P<0.05);显著降低pH、全盐含量和土壤容重,降幅分别为8.9%、27.9%和17.9%;显著提升土壤速效磷和速效钾指标,增幅分别为36.0%和17.4% (P<0.05)。【结论】GBW HF-98菌株具有较强的胞外多糖产生能力,在中度盐碱土壤中促进番茄生长和改良土壤的效果显著,可用于盐碱土壤改良菌剂的研制。     

假单胞菌胞外多糖发酵条件的研究

研究了假单胞菌（Ｐｓｅｕｄｏｍｏｎａｓｓｐ．６）利用木糖产胞外多糖的发酵条件。实验表明ＫＨ２ＰＯ４,Ｏ２和高碳氮比对多糖合成有促进作用,在发酵后期补加木糖有助于多糖产量的提高     

Biosynthesis, characterisation, and design of bacterial exopolysaccharides from lactic acid bacteria

Lactic acid bacteria (LAB) are characterised by their conversion of a large proportion of their carbon feed, fermentable sugars, to lactic acid. However, in addition to lactic acid production, the LAB are able to divert a small proportion of fermentable sugars towards the biosynthesis of exopolysaccharides (EPSs) that are independent of the cell surface and cell wall material. These microbial EPSs when suspended or dissolved in aqueous solution provide thickening and gelling properties, and, as such, there is great interest in using EPSs from food grade microorganisms (such as the LAB that are traditionally used for food fermentations) for use as thickening agents. The current review includes a brief summary of the recent literature describing features of the biosynthetic pathways leading to EPS production. Many aspects of EPS biosynthesis in LAB are still not fully understood and a number of inferences are made regarding the similarity of the pathway to those involved in the synthesis of other cell polysaccharides, e.g., cell wall components. The main body of the review will cover practical aspects concerned with the isolation and characterisation of EPS structures. In the last couple of years, a substantial number of structures have been published and a summary of the common elements of these structures is included as is a suggestion for a system for representing structures. A brief highlight of the attempts that are being made to design ‘tailor’-made polysaccharides using genetic modification and control of metabolic flux is presented.     

Shear treatment of starter culture medium improves separation behavior of Streptococcus thermophilus cells

A central step in the production of starter cultures is the separation of the cells from the fermentation medium, which is usually achieved by disk centrifuges. In case of microorganisms which produce exopolysaccharides (e.g., various strains of lactic acid bacteria), the properties of the respective exopolysaccharides may interfere with this separation step. By using six strains of Streptococcus thermophilus the hypothesis was tested that a shear treatment of the fermented culture medium improves subsequent cell separation markedly. Depending on the type of exopolysaccharides (freely present in the medium, or as capsules around the cells) an energy input of up to 2.5 kJ/mL generated with an Ultra‐Turrax affected cell chain length of the strains and viscosity of fermentation medium differently. For bacteria producing capsular exopolysaccharides, space‐ and time‐resolved centrifugation experiments revealed an increase of sedimentation velocity after shear treatment. In general, viability of the microorganisms, detected by flow cytometry measurements and fermentation experiments, was not affected by the shearing procedure. The results therefore indicate that strain‐targeted shearing is helpful to improve the separability of cells from the fermented media.     

Viili中益生菌的分离及其发酵性质研究

目的 利用传统方法和分子生物学方法从viili中筛菌并对其特性进行研究.方法 采用MRS、YPD和脱脂乳营养琼脂3种平板从viili中分离出15株单菌,利用V3区通用引物和乳杆菌特异性引物对各单菌的PCR产物进行变性梯度凝胶电泳(denaturing gradient gel electrophoresis,DGGE)分析,将其归为5种不同菌.结果 16S DNA测序表明,5种单菌分别为Lactobacillus plantarum、Streptococcus thermophilus、Lactobacillus paracasei、Bacillus cereus和Lactobacillus delbrueckii subsp. bulgaricus.混合发酵实验表明:从viili中筛出的5种菌除Bacillus cereus外均有凝乳现象,其中Lactobacillus delbrueckii凝乳能力强,可在5 h内凝乳;Lactobacillus paracasei、Bacillus cereus和Lactobacillus plantarum组合产生的EPS量最多,高达186.71 mg/L,而Streptococcus thermophilus EPS产量仅为33.56 mg/L.结论 传统方法与分子生物学方法DGGE相结合,可以快速准确判断细菌种类;筛选菌特性研究结果表明,细菌之间的相互作用导致凝乳时间和EPS产量发生变化,其复合作用有待于进一步研究.     

Potentials of Exopolysaccharides from Lactic Acid Bacteria

Recent research in the area of importance of microbes has revealed the immense industrial potential of exopolysaccharides and their derivative oligosaccharides from lactic acid bacteria. However, due to lack of adequate technological knowledge, the exopolysaccharides have remained largely under exploited. In the present review, the enormous potentials of different types of exopolysaccharides from lactic acid bacteria are described. This also summarizes the recent advances in the applications of exopolysaccharides, certain problems associated with their commercial production and the remedies.     

元阳豆豉中高产蛋白酶乳酸菌的筛选及其产酶条件的研究

从云南省元阳地区采集豆豉样品,并从中分离得到62株乳酸菌菌株。通过脱脂乳平板试验,从中初步筛得到21株具有高蛋白酶活力的菌株,采用茚三酮法测定其蛋白酶活力,复筛出高产蛋白酶菌株YY-1-6L,并对其产酶条件进行研究。结果表明,YY-1-6L在以葡萄糖为碳源、多聚蛋白胨为氮源、起始pH 5.0的明胶诱导培养基中,接种量为3%,35℃发酵培养36 h,其蛋白酶活力高达32.50 U/mL,且K2HPO4、MgSO4能促进蛋白酶产生。     

产生物胺乳酸菌的筛查与检测

目的对上海市场上食品和药品中分离出的20株乳杆菌,13株链球菌,3株乳球菌和3株肠球菌的产生物胺能力进行检测,以揭示其潜在的安全性问题。方法检测的生物胺共包括6种：分别为酪胺、精胺、尸胺、组胺、腐胺和色胺。利用添加了前体氨基酸的氨基酸脱羧酶筛选培养基对各菌株的产胺能力进行初筛,通过培养基中指示剂的颜色变化判定产胺能力。结果在检测的39株菌中,8株菌具有产酪胺的能力,7株菌具有产精胺的能力,1株菌具有产组胺的能力,1株菌具有产腐胺的能力。尤其是精胺和酪胺的产量较为引人关注。结论生物胺的危害水平取决于个体解毒的能力,但在筛选食品药品用菌株时应运用规范的方法来检测其产生物胺的能力,以保障相应食品药品的安全问题。     

富锌产γ-氨基丁酸乳酸菌的筛选及初步鉴定

用含0.2%-0.25%Zn^2＋的MRS培养基从发酵酸菜、酸奶中分离出6株具富锌能力细菌,经形态和生理生化特性鉴定,初步判断菌株为乳酸菌。采用纸层析检测MRSG培养基培养的6株菌上清液后,初步确定有4株能产γ-氨基丁酸。以此4菌株基因组为模板PCR扩增到540 bp的片段,经测序和序列分析,证明是谷氨酸脱羧酶基因高度保守区域,初步证明谷氨酸脱羧酶基因的存在。此4株菌发酵液经HPLC检测和分析后,证实2号菌株能转化谷氨酸钠生成GABA,产量为4.8 g/L,在应用方面具有很大的潜力。     

Bioprospecting of indigenous resources for the exploration of exopolysaccharide producing lactic acid bacteria

Exploration of biodiversity lead towards the discovery of novel exopolysaccharide (EPS) producing microbes that have multiple applications. The safety compatibility status of lactic acid bacteria (LAB) makes it an attractive candidate for the production of EPS in industries. Therefore, new bacterial isolates are continuously being identified from different habitats. Current research was conducted to explore indigenous biodiversity for the production of dextransucrase, which is involved in the synthesis of dextran. Dextran is an EPS which is used in different industries. In this study, thirty-nine LAB were isolated from different food samples. The isolates were identified as genus Leuconostoc, Weissella and Streptococcus based on genotypic and phenotypic characteristics. Screening revealed that only eight isolates can produce dextransucrase in high titres. Fermentation conditions of dextran producing LAB was optimized. The results indicated that Weissella confusa exhibited maximum specific activity (1.50?DSU?mg^?1) in 8?h at 25?°C with pH 7.5. Dextran produced from Weissella proved to be a useful alternative to commercially used dextran produced by Leuconostoc mesenteroides in industries for various applications.     

高效降解草酸乳酸菌的筛选

尿路结石70%~80%主要由草酸钙结晶构成。人体内的草酸一般通过肠道内微生物降解,经由粪便排出或在泌尿道吸收由尿液排出。本研究对市场上商品化的发酵乳制品、饮料和药品中的乳酸菌进行分离,得到37株菌,包括嗜热链球菌、保加利亚乳杆菌、干酪乳杆菌、鼠李糖乳杆菌、植物乳杆菌、嗜酸乳杆菌、动物双歧杆菌、长双歧杆菌、粪肠球菌、屎肠球菌和乳球菌,并检测这些菌株降解草酸的能力。结果提示,乳酸菌在体外能够有效的降低培养物中的草酸浓度,并筛选出了具有高效降解草酸能力的乳酸菌菌株。有望成为尿石症预防的新措施。     

植酸酶产生菌的筛选与酶纯化及其性质的研究

对从土壤中分离得到的一株产植酸酶的细菌进行了生理生化鉴定,并对植酸酶进行了分离纯化,该酶反应的最适温度约为55℃,最适pH值为5．8,植酸酶蛋白分子量约为14kD。