乳酸菌胞外多糖的研究进展

乳酸菌胞外多糖是乳酸菌在生长代谢过程中分泌到细胞壁外、常渗于培养基中的一类天然的生物高分子聚合物。由于胞外多糖具有降低胆固醇、抗肿瘤、促进菌体在肠粘膜上的粘附和免疫应答以及可作为食品增稠剂和稳定剂等重要的生理和物理特性而成为近年来的研究热点。了解乳酸菌胞外多糖的合成途径、结构以及构效关系,对于提高乳酸菌胞外多糖产量、深入了解其生物活性相关机制具有重要的意义。综述了乳酸菌胞外多糖的分类、构效关系、生物合成机制以及生物活性。     

Immunomodulatory and antioxidant potential of Lactobacillus exopolysaccharides

BACKGROUND: Immunomodulation by probiotic microorganisms has become a topic of increasing interest in food microbiology. Polysaccharides are broadly used in the food industry as gelling, thickening, stabilizing, or emulsifying agents. Some probiotics such as lactic acid bacteria also produce exopolysaccharides that stimulate macrophage production of cytokines. The aim of this study was to characterize the effects of exopolysaccharides of Lactobacillus paracasei subsp. paracasei NTU 101 (101EP) and Lactobacillus plantarum NTU 102 (102EP) exopolysaccharides on antioxidant activity and immunomodulation in vitro. RESULTS: The sugar composition (including arabinose, galactose, glucose, fructose, mannose, and maltose) of 101EP and 102EP was quantified by high‐performance anion‐exchange chromatography. Cytokine production (including IL‐6, TNF‐α, and IL‐1β) was induced by 101EP and 102EP in Raw 264.7 in a dose‐dependent manner (5‐500 µg mL^?1). 101EP and 102EP also demonstrated potential antioxidant properties (1,1‐diphenyl‐2‐picrylhydrazyl radical scavenging activity, chelation of ferrous ions, inhibition of linoleic acid peroxidation, and reducing power) in vitro. CONCLUSION: 101EP and 102EP stimulate cell proliferation and may be useful as a mild immune modulator of macrophages. Copyright © 2011 Society of Chemical Industry     

乳酸菌胞外多糖特性及其合成影响因素研究进展

介绍了乳酸菌胞外多糖的分类、化学组成、结构特性和遗传调控的研究进展。对乳酸菌胞外多糖的合成菌株、底物、温度、pH值和添加物等多种影响因素的研究成果进行了综述，并对不同菌株在不同的合成条件下的生物合成量进行了归纳，以期对国内开展有关方面的研究提供一定的参考和帮助。     

Interactions between milk proteins and exopolysaccharides produced by Lactococcus lactis observed by scanning electron microscopy

The interactions between exopolysaccharides produced by Lactococcus lactis ssp. cremoris JFR1 and dairy proteins (caseins and whey proteins) in fermented media (milk permeate and buttermilk) were observed using scanning electron microscopy. An immobilization technique by crosslinking was employed to bind the protein to the observation surface, so that a washing step could be performed to remove noninteracting material. The use of this novel technique allowed us, for the first time, to confirm that the exopolysaccharide molecules interact with dairy proteins. Exopolysaccharides appear as filament strands attached to the protein aggregates and to the bacterial cells. This new sample preparation technique proved to be very valuable for observing molecular interactions in fermented media.     

Invited review: methods for the screening, isolation, and characterization of exopolysaccharides produced by lactic acid bacteria

The ability to produce exopolysaccharides (EPS) is widespread among lactic acid bacteria (LAB), although the physiological role of these molecules has not been clearly established yet. Some EPS confer on LAB a "ropy" character that can be detected in cultures that form long strands when extended with an inoculation loop. When EPS are produced in situ during milk fermentation they can act as natural biothickeners, giving the product a suitable consistency, improving viscosity, and reducing syneresis. In addition, some of these EPS may have beneficial effects on human health. The increasing demand by consumers of novel dairy products requires a better understanding of the effect of EPS on existing products and, at the same time, the search for new EPS-producing strains with desirable properties. The use of genetically modified organisms capable of producing high levels of EPS or newly designed biopolymers is still very limited. Therefore, exploration of the biodiversity of wild LAB strains from natural ecological environments is currently the most suitable approach to search for the desired EPS-phenotype. The screening of ropy strains and the isolation and characterization of EPS responsible for this characteristic have led to the application over the past years of a wide variety of techniques. This review summarizes the available information on methods and procedures used for research on this topic. The information provided deals with methods for screening of EPS-producing LAB, detection of the ropy phenotype, and the physicochemical and structural characterization of these molecules, including parameters related to their viscosifying properties. To our knowledge, this is the first compilation of methods available for the study of EPS produced by LAB.     

Comparison of the functionality of exopolysaccharides produced in situ or added as bioingredients on yogurt properties

The effect on yogurt properties of in situ production of exopolysaccharides (EPS) and addition of 2 EPS powders (crude and purified EPS from Lactobacillus rhamnosus RW-9595M fermentation in whey-based medium) at different concentrations was studied. No effect of purified powder addition for EPS concentrations up to 500 mg/L was observed on acidification rate to the difference of milks supplemented with crude EPS, which exhibited longer acidification times. The addition of EPS from 125 to 500 mg/L or the use of EPS-producing cultures resulted in yogurts with lower yield stress and viscoelastic moduli compared with control yogurts without EPS, with no apparent effect of EPS concentration. However, the consistency index was higher for yogurts produced with the commercial EPS-producing culture, and to a lesser extent with the mixed culture containing Lb. rhamnosus RW-9595M, compared with yogurts supplemented with EPS powders, which were not different from that for control yogurts. Our study showed that the mode of EPS incorporation in yogurts has a major effect on the rheological properties of the final product.     

鼠李糖乳杆菌JAAS8胞外多糖生物合成基因的克隆和序列分析

根据已报道乳杆菌胞外多糖生物合成基因的同源性,利用其保守区设计引物,扩增出鼠李糖乳杆菌JAAS8RmlA基因部分序列,并通过染色体步移法扩增出Rml(ACB)的序列。获得了鼠李糖乳杆菌JAAS8胞外多糖生物合成基因4.1 kb序列片段,编码4个可读框。利用BLAST和ClustalX程序对获得序列进行生物信息学分析,预测其结构和功能。结果表明:该序列与已知鼠李糖乳杆菌胞外多糖生物合成基因具有高度同源性,RmlA、RmlC和RmlB基因与dTDP-L-鼠李糖前体的生物合成密切相关。     

丝状真菌在纤维素酶合成过程中碳源代谢调控的研究进展

丝状真菌可以分泌大量纤维素酶并能有效地降解纤维素底物。在纤维素酶的合成过程中,碳源对产酶微生物有着不可替代的作用,然而对于微生物碳源响应及代谢调控有关的机理仍不是很清楚,从而制约了纤维素酶产业的发展。因此,本文综述了丝状真菌中转录调控因子对纤维素酶合成过程的影响,探讨了真菌在纤维素酶表达过程中对碳源代谢的调控,包括对碳源的响应、碳源对产酶过程的诱导阻遏,并展望了纤维素酶的研究方向。     

动物双歧杆菌乳亚种M8胞外多糖的分离纯化和分子特征

动物双歧杆菌是一种高产胞外多糖(exopolysaccharide,EPS)的益生菌,在食品工业中应用广泛.该研究以动物双歧杆菌亚种M8为研究对象,发酵提取EPS,分别选用乙醇分级沉淀(ethanol gradient precipitation,EGP)法和阴离子交换色谱(anion-exchange chromat...     

Use of Lactobacillus paracasei isolated from whey for silver nanocomposite synthesis: Antiradical and antimicrobial properties against selected pathogens

The present research emphasizes the use of safe, inexpensive, and available whey using Lactobacillus paracasei as a source in silver nanocomposite synthesis as an alternative bioactive agent for dairy and biomedical applications. Through the multiinstrumental approach used in this study based on spectroscopic and microscopic methods as well as spectrometric techniques, the characterization and evaluation of silver composites and their antimicrobial and antiradical properties were enabled. Synthesized silver nanocomposites have been found in form of nanocrystals, naturally coated by an organic surface with high antimicrobial and antiradical properties. Furthermore, this work also presents an innovative approach regarding the organic surface (naturally secreted by the bacteria isolated from whey) of the core of nanoparticles, which has already been explored and therefore is starting to supplement the scientific approach concerning biologically synthesized nanoparticles. This work also presents a general frame on the resistance subject by performing the trial interaction of commercially available antibiotics (kanamycin and ampicillin) with new bioactive compounds that can create novel knowledge on complementing their action. Moreover, synthesized silver nanocomposites have shown great antioxidant and antimicrobial effects against various foodborne pathogens from dairy products and drug resistance pathogens found in the medical area to rank on the top of mortality rate.     

鼠李糖乳杆菌JAAS8胞外多糖生物合成基因的克隆及生物信息学分析

根据已知乳杆菌胞外多糖生物合成基因的同源性,利用其保守区设计引物,扩增出鼠李糖乳杆菌JAAS8磷蛋白磷酸酶基因(wzb)序列,并通过染色体步移法克隆了鼠李糖乳杆菌 JAAS8 参与胞外多糖生物合成基因簇全部序列 (19.7kb),利用生物信息学方法预测了基因簇 16个阅读框的结构和功能。结果表明,该序列与已报道的乳杆菌胞外多糖生物合成基因具有高度同源性。welF、welG、welH、welI、welJ和welE为合成寡糖重复单元的糖基转移酶基因。rmlA、rmlB和rmlC基因与dTDP-L-鼠李糖前体的生物合成密切相关。wzd、wze、wzy、wzx、wzr和wzb基因预测蛋白主要参与胞外多糖合成过程中多糖链长检测、聚合和输出。     

Exopolysaccharides produced by Lactobacillus sp.: Biosynthesis and applications

Lactobacillus sp. synthesize exopolysaccharides (EPS), including both homo- and heteropolysaccharides, which play an important role in the production of fermented foods, and especially in the dairy industry, improving the gustatory and rheological properties of the finished products. These polymers are generated by starter cultures in situ in fermented foods, and so they are treated as natural thickening agents. As some Lactobacillus strains are generally recognized as safe and have been shown to exhibit probiotic activity, EPS from those bacteria can be used as functional food ingredients, conferring both health and economic benefits to the consumers. However, their industrial applications are hindered by the low yield of EPS from Lactobacillus and high costs of their purification. This review focuses on the latest reports concerning the biosynthesis and properties of Lactobacillus EPS.     

Characterization and antimicrobial activity of bacteriocin 217 produced by natural isolate Lactobacillus paracasei subsp. paracasei BGBUK2-16

The strain Lactobacillus paracasei subsp. paracasei BGBUK2-16. which was isolated from traditionally homemade white-pickled cheese, produces bacteriocin 217 (Bac217; approximately 7 kDa). The onset of Bac217 biosynthesis was observed in the logarithmic phase of growth, and the production plateau was reached after 9 or 12 h of incubation at 37 and 30 degrees C, respectively, when culture entered the early stationary phase. Biochemical characterization showed that Bac217 retained antimicrobial activity within the range of pH 3 to 12 or after treatment at 100 degrees C for 15 min. Bac217 antimicrobial activity also remained unchanged after storage at 4 degrees C for 6 months or -20 degrees C for up to 12 months. However, Bac217 activity was completely lost after treatment with different proteolytic enzymes. BGBUK2-16 contains only one plasmid about 80 kb in size. Plasmid curing indicated that genes coding for Bac217 synthesis and immunity seem to be located on this plasmid. Bac217 exhibited antimicrobial activity against some pathogenic bacteria, such as Staphylococcus aureus and Bacillus cereus. Interestingly, Bac217 showed activity against Salmonella sp. and Pseudomonas aeruginosa ATCC27853. The inhibitory effect of BGBUK2-16 on the growth of S. aureus in mixed culture was observed. S. aureus treatment with Bac217 led to a considerable decrease (CFU/ml) within a short period of time. The mode of Bac217 action on S. aureus was identified as bactericidal. It should be noted that the strain BGBUK2-16 was shown to be resistant to bacteriocin nisin, which is otherwise widely used as a food additive for fermented dairy products.     

乳酸菌产胞外多糖的研究

综述了乳酸菌产胞外多糖的研究进展，包括产胞外多糖的菌株、影响生物合成的因素等，并介绍了乳酸菌产胞外多糖的开发及应用。     

Structural and technological characterization of ropy exopolysaccharides produced by Lactobacillus plantarum strains isolated from Tarhana

Food Science and Biotechnology - Exopolysaccharide producing starter cultures enable manufacturing “clean labeled” foods with improved textural and nutritional properties. The...     

Isolation and partial biochemical characterization of a proteinaceous anti-bacteria and anti-yeast compound produced by Lactobacillus paracasei subsp. paracasei strain M3

New proteinaceous active substance produced by Lactobacillus paracasei subsp. paracasei strain M3 used as a starter for Bulgarian yellow cheese was identified and studied. It displayed bactericidal and fungistatic activities. Its activity was checked against over 60 bacterial and yeast strains. It was efficient against Bacillus subtilis ATCC 6633, several L. delbrueckii species, Helicobacter pylori NCIPD 230 and some yeast species, for example Candida albicans, C. pseudointermedia NBIMCC 1532, C. blankii NBIMCC 85 and Saccharomyces cerevisiae NBIMCC 1812. The synthesis of the substance by producing strain was detected in the late logarithmic growth phase during batch fermentation. Anion exchange chromatography, reversed phase chromatography (RPC) on C4 column and HPLC on C18 column were used for partial purification of this antimicrobial compound. The gene responsible for the synthesis of the active substance is located on the bacterial chromosome.     

不同碳源条件下产色素甲烷氧化细菌的菌株特性研究

甲烷氧化菌能利用甲烷作为唯一的碳源和能源。对甲烷氧化细菌M13在甲烷、甲醇不同碳源下进行培养,测定生长曲线,并对菌落形态、细胞形态和菌种生长情况进行观察和分析。结果表明,甲烷氧化细菌M13为革兰氏阴性菌,细胞呈短杆状或略弯,该菌菌落较大,呈圆型,乳白色,较为均匀。长期培养能产生粉红色素,以甲醇作碳源比甲烷作碳源的对菌体生长更好。      

Binding of mutagens to exopolysaccharide produced by Lactobacillus plantarum mutant strain 301102S

Exopolysaccharide (EPS) was produced by Lactobacillus plantarum 301102 on exposure to the mutagenic action of acridine orange and novobiocin. The biological characteristics of this mutant strain 301102S were the same as those of the parent strain, but fermented milk prepared with the mutant strain showed antimutagenic activity on 3-amino-1,4-dimethyl-5H-pyrido indole. Only EPS-bound cells of strain 301102S showed binding ability to mutagens such as heterocyclic amines, and the mutagens were inactivated by binding to EPS. The binding ability was affected by pH; the greatest percentage binding was noted at pH 8.0. Addition of Mg²⁺ and sodium dodecyl sulfate, but not oxgall, inhibited the binding ability. Therefore, the binding mechanism of the EPS may consist of ion-exchange and hydrophobic bonds, and the EPS would bind mutagens in the intestine.     

丙酸杆菌发酵条件的优化

讨论pH值和不同碳源对丙酸杆菌的影响,同时利用正交实验优化了此菌发酵的培养基。