Structural and rheological characterisation of heteropolysaccharides produced by lactic acid bacteria in wheat and sorghum sourdough

Hydrocolloids improve the volume, texture, and shelf life of bread. Exopolysaccharides (EPS) produced by lactic acid bacteria (LAB) during sourdough fermentation can replace hydrocolloids. It was the aim of this study to determine whether heteropolysaccharides (HePS) synthesized intracellularly from sugar nucleotides by glycosyltransferases are produced in wheat and gluten-free sorghum sourdough at effective levels. The HePS-producing strains Lactobacillus casei FUA3185, L. casei FUA3186, and Lactobacillus buchneri FUA3154 were used; Weissella cibaria 10M producing no EPS in the absence of sucrose served as control strain. Cell suspensions of L. buchneri in MRS showed the highest viscosity at low shear rate. Glycosyltransferase genes responsible of HePS formation in LAB were expressed in sorghum and wheat sourdough. However, only HePS produced by L. buchneri influenced the rheological properties of sorghum sourdoughs but not of wheat sourdoughs. Sorghum sourdough fermented with L. buchneri exhibited a low |G^∗| compared to the control, indicating a decrease in resistance to deformation. An increase in tan δ indicated decreased elasticity.The use of LAB producing HePS expands the diversity of EPS and increases the variety of cultures for use in baking.     

木质素热化学解聚与协同解聚的进展

对木质素解聚方法、解聚过程中所使用的催化剂和溶剂以及解聚工艺条件等进行了综述,总结了各种解聚方式的优缺点,同时阐述了协同解聚木质素。展望了木质素解聚发展方向。     

Super-channel in bacteria: structural and functional aspects of a novel biosystem for the import and depolymerization of macromolecules

Cells of Sphingomonas sp. A1 directly incorporate a macromolecule, alginate, into the cytoplasm through a biosystem, super-channel, consisting of a pit on the cell surface, alginate-binding proteins in the periplasm, and an ATP-binding cassette transporter in the inner membrane. The alginate is finally depolymerized into constituent monosaccharides by polysaccharide lyases present in the cytoplasm. The fundamental frame of the biosystem for alginate transport, and the functions of the pit, binding proteins, and ABC transporter have already been reviewed together with those of alginate-depolymerization processes [Hashimoto et al., J. Biosci. Bioeng., 87, 123-136 (1999)]. In this review, we have attempted to demonstrate the three-dimensional structure and evolution features of the super-channel, and alginate-depolymerization processes by using information obtained mainly through genomics, proteomics, and X-ray crystallography.     

Enzymatic hydrolysis of soybean protein using lactic acid bacteria

The proteolytic activity of 12 lactic acid bacteria (LAB) strains, assayed on soy protein extract at a temperature of 37 °C for 6 h, was evaluated by SDS–PAGE, reverse-phase HPLC and free-amino acid analyses. The results indicated that α- and α′-subunits of β-conglycinin were the preferred substrates for the majority of the LAB. Only a few strains exerted some action against the basic polypeptides of glycinin, this fraction was the least degraded of all soy protein fractions. Whole-cell suspensions of LAB used in this study generated hydrophilic and hydrophobic peptides from mainly soy protein fractions. RP-HPLC analyses indicated differences in the profiles of the hydrolysates, with several peaks decreasing in size and new peaks being formed. Three of the selected strains assayed increased the level of total free amino acids in the soy protein extract (SPE) and hydrolyzed principally essential amino acids and flavour precursor amino acids.     

Electrokinetic transport of PAH-degrading bacteria in model aquifers and soil

An investigation of the mobility, viability, and activity of polycyclic aromatic hydrocarbon (PAH) degrading bacteria in an electric field is presented. Bench-scale model aquifers were used to test electrophoresis and electroosmosis as potential mechanisms for bacterial dispersion in contaminated sites. Glass beads, alluvial sand from Lake Geneva, and historically polluted clayey soil were used as packing materials. The green-fluorescent protein labeled PAH-degrading bacteria Sphingomonas sp. L138 and Mycobacterium frederiksbergense LB501TG were used as test organisms because of the known differing physicochemical surface and adhesion properties of the corresponding wild-type strains. No adverse effects of the electric current on bacterial viability and PAH-degradation were observed in the system chosen. Up to 90% of the weakly negatively charged and moderately adhesive cells of strain L138 were transported by electroosmosis, whereas 0-20% were transported by electrophoresis. By contrast, poor electrokinetic transport of strongly charged and highly adhesive cells of M. frederiksbergense LB501TG occurred in the different model aquifers. Treatment of bacteria with the nonionic surfactant Brij35 resulted in up to 80% enhanced electrokinetic dispersion of both strains. Our findings demonstrate that electroosmosis may be a valuable mechanism to transport bacteria in the subsurface with transport efficiencies heavily depending on the retention of the bacteria by the solid phase.     

高效同化胆固醇乳酸菌菌株的筛选及在食品中的应用

从发酵食品中分离筛选能高效同化胆固醇的乳酸菌,以开发低胆固醇系列产品。从25个来源不同的样品中,分离筛选出8株乳酸菌,与活化出的8株已知乳酸菌共同在特殊培养基中培养,最后筛选出2株具有较高同化胆固醇能力的菌株。结果表明,嗜酸乳杆菌和植物乳杆菌应用于食品中,对食品风味影响较小,可大量应用。     

预拓扑空间的邻域系、开邻域基与基

定义了预拓扑空间的邻域系、开邻域基与基;研究了邻域系和开邻域基的性质;给出了两个开邻域基算子彼此等价的充分必要条件以及两个预拓扑基彼此等价的充分必要条件;讨论了预拓扑空间的可数基的一些性质;证明了有限预拓扑有最小基并给出了求最小基的3种算法．     

Strategies of nutrient transport by ruminal bacteria

The survival of bacteria in natural environments like the rumen depends on the ability of the bacteria to scavenge nutrients. It is now evident that ruminal bacteria use a variety of transport mechanisms. Hydrophobic substances, such as ammonia and acetate, are permeable to the lipid bilayers of cell membranes and can be taken up by passive diffusion. Hydrophilic compounds (e.g., sugars, amino acids, peptides) do not easily pass through lipid bilayers and must be transported across cell membranes on carrier proteins. Facilitated diffusion can display saturable kinetics but does not result in accumulation of solute. Active transport can establish extremely high concentration gradients, and this work may be driven by the hydrolysis of chemical bonds (e.g., ATP) or ion gradients, which are coupled to solute symport. Many solute symports involve protons, but sodium systems also are common in ruminal bacteria. The phosphotransferase system chemically modifies sugars as they pass across the cell membrane, and several ruminal bacteria have this method of group translocation. Many feed additives have either a direct or indirect effect on rumen bacterial transport. For instance, ionophores can inhibit transport by destroying (sometimes even reversing) ion gradients, lowering intracellular pH, or causing excessive ATP hydrolysis.     

海藻德合酶基因工程菌诱导表达条件的研究

实验将构建后的重组质粒pET21TreS转入大肠杆菌Rosetta gami(DE3)中,得到海藻糖合酶基因工程菌,并通过对培养时间、温度及诱导剂浓度等条件的优化,对其进行诱导表达,得到了该基因工程菌诱导表达的最适条件:发酵培养至菌体浓度(OD_(600))至0.6～0.8时,以1mmol/L浓度的异丙基-β-D-硫代半乳糖苷(IPTC),37℃,诱导2h后得到的蛋白表达量较高.     

细菌原生质体融合育种技术及其应用进展

原生质体融合技术是细菌遗传育种的有效方法之一,发展迅速,应用广泛。文中综述了亲本菌株选择性遗传标记方法、影响原生质体制备与再生因素、原生质体融合方法和条件。介绍了细菌原生质体融合技术在遗传育种中的应用,并展望了细菌原生质体融合技术的发展前景。     

Antibacterial activity and cholesterol assimilation of lactic acid bacteria isolated from traditional Iranian dairy products

In this study, lactic acid bacteria were isolated from ewe milk, traditional yoghurt and sour buttermilk samples collected from different areas of Azarbayjan-e-sharqi in Iran. All the isolates were screened for their ability to produce bacteriocin like inhibitory substances (BLIS) by studying their inhibitory action against pathogens like Listeria monocytogenes, Salmonella enteritidis and Staphylococcus aureus, after eliminating the effect of organic acids and hydrogen peroxide. According to results, four of the isolates identified as Lactobacillus brevis, Lactobacillus pentosus, Pedoicoccus acidilactici and Lactobacillus paracasei were unaffected by the action of pH neutralization and hydrogen peroxide and showed inhibitory action against the tested pathogens. The inhibitory activities demonstrated by these isolates were completely inhibited in the presence of proteolytic enzymes.     

Fecal indicator bacteria transport and deposition in saturated and unsaturated porous media

Beach sediment and sand are recognized as nonpoint fecal indicator bacteria (FIB) sources capable of causing water quality and health risks for beach-goers. A comprehensive understanding of the key factors and mechanisms governing the migration and exchange of FIB between beach water column and sediment is desired to better predict FIB concentration variations and assess the associated risk. The transport and retention behavior of two model FIB Enterococcus faecalis (E. faecalis) and Escherichia coli (E. coli) was examined using packed-bed columns in both saturated and unsaturated porous media to evaluate FIB migration potentials at conditions simulating the coastal aquatic environment. Additionally, complementary cell characterization techniques were conducted to better understand the migration behaviors of both FIB strains observed in the column experiments. The mobility of the gram-positive species E. faecalis was much more sensitive to solution chemistry and column saturation level than that of the gram-negative species E. coli. Interaction energy calculations suggest that E. faecalis retention was largely governed by the combination of DLVO (Derjaguin-Landau-Verwey-Overbeek) and non-DLVO (most likely hydrophobic and/or polymer bridging) interactions in saturated porous media, while the combination of DLVO and steric interactions controlled the deposition of E. coli cells. The measured surface properties of the two FIB strains supported the distinct bacteria transport behaviors and the differences of the identified mechanisms for each strain. As a result, E. faecalis showed the least affinity to sand in freshwater and appeared to be irreversibly attached in primary energy minima at elevated salt conditions; whereas the retained E. coli cells were reversibly attached and mostly associated with the secondary energy minima at both freshwater and seawater conditions. In unsaturated porous media, E. faecalis cells seemed to prefer to attachment at air/water interface rather than sand surface, while E. coli showed a similar affinity to the two interfaces. It was proposed that the different surface characteristics of the two FIB strains resulted in the distinct transport and retention behavior in porous media. These results highlight the need for FIB management to consider variations in transport behavior between model FIB when assessing water quality and associated risks.     

Bacteriophage-derived genetic tools for use in lactic acid bacteria

Strains of lactic acid bacteria (LAB) are commonly used in industrial fermentations for the production of food products such as cheeses, buttermilk, sauerkraut and yogurt. It has long been recognised that bacteriophages infecting LAB strains can cause serious problems in dairy fermentations, resulting in economic losses and therefore research has been focussed on elucidating the mechanisms by which these bacteriophages proliferate. In recent years, significant advances in molecular biological technologies has allowed researchers to gain an insight into the genetic processes underlying many steps in LAB bacteriophage lifecycles. The knowledge gained from this research has in turn, enabled the development of novel genetic tools utilising phage genes, promoters, and DNA fragments for use in LAB strains, which is the focus of this review.     

Enzymatic activities of lactic acid bacteria isolated from Cornetto di Matera sourdoughs

Forty-one strains of lactic acid bacteria (LAB) isolated from Cornetto di Matera sourdoughs were screened for their enzymatic activities, to elucidate their possible roles during the fermentation process. Urease, peptidase, phytase, phosphatase and beta-glucosidase activities were measured spectrophotometrically using synthetic substrates. Proteolytic activities were examined in model doughs, using neutral and acidified sterile doughs as controls. All strains had low urease, glutamyl aminopeptidase and iminopeptidase activities, whereas differences within species were observed for the other enzymatic activities. Leuconostoc mesenteroides and Lactobacillus curvatus strains generally showed high aminopeptidase, X-prolyl dipeptidyl aminopeptidase, beta-glucosidase and phytase activities, while the enzymatic activities of Lactobacillus plantarum, Lactobacillus pentosus and Weissella cibaria varied between strains. In order to classify the strains on the basis of similar enzymatic profiles, a hierarchical cluster analysis was carried out. Several strains of L. plantarum, L. curvatus and Leuc. mesenteroides showed an interesting combination of proteolytic, peptidase, beta-glucosidase and phytase activities, suggesting their possible usefulness as a mixed starter culture in bread-making processes.     

Isolation and genetic identification of spore-forming bacteria associated with concentrated-milk processing in Nebraska

Spore-forming bacteria are heat-resistant microorganisms capable of surviving and germinating in milk after pasteurization. They have been reported to affect the quality of dairy products by the production of enzymes (lipolytic and proteolytic) under low-temperature conditions in fluid milk, and have become a limiting factor for milk powder in reaching some selective markets. The objective of this research was to isolate and identify the population of spore-forming bacteria (psychrotrophic and thermophilic strains) associated with concentrated milk processing in Nebraska. During 2 seasons, in-process milk samples from a commercial plant (raw, pasteurized, and concentrated) were collected and heat-treated (80°C/12 min) to recover only spore-formers. Samples were spread-plated using standard methods agar and incubated at 32°C to enumerate mesophilic spore counts. Heat-treated samples were also stored at 7°C and 55°C to recover spore-formers that had the ability to grow under those temperature conditions. Isolates obtained from incubation or storage conditions were identified using molecular techniques (16S or rpoB sequencing). Based on the identification of the isolates and their relatedness, strains found in raw, pasteurized, and concentrated milk were determined to be similar. Paenibacillus spp. were associated with both raw and concentrated milk. Due to their known ability to cause spoilage under refrigeration, this shows the potential risk associated with the transferring of these problematic organisms into other dairy products. Other Bacillus species found in concentrated milk included Bacillus clausii, Bacillus subtilis, Lysinibacillus sp., Bacillus safensis, Bacillus licheniformis, Bacillus sonorensis, and Brevibacillus sp., with the last 3 organisms being capable of growing at thermophilic temperatures. These strains can also be translocated to other dairy products, such as milk powder, representing a quality problem. The results of this research highlight the importance of understanding spore-formers associated with the processing of condensed milk, which then may allow for specific interventions to be applied to control these microorganisms in this processing chain. To our knowledge, this is the first study evaluating spore-formers associated with concentrated milk in the United States.     

Effect of soybean proteins on gluten depolymerization during mixing and resting

BACKGROUND: Gluten and soy proteins interact as a consequence of dough mixing; however, there is no evidence of the effect of soy protein on gluten depolymerization. The aim of this study was to assess the depolymerizing effect of soy protein on gluten network after mixing and resting of mixed doughs. Therefore, the changes in glutenin macropolymer (GMP) content, protein composition and free sulfhydryl content were evaluated. RESULTS: The protein profile from gluten–soybean blends, obtained by multistacking SDS‐PAGE, showed differences when compared to gluten profile. Soy and gluten proteins were extracted together with SDS buffer, which showed that soy proteins remained associated to insoluble wheat proteins even after hand‐washing the dough to obtain gluten. GMP content was determined to analyze the effect of soy protein on GMP gel formation. Protein content of GMP obtained from flour mixes and doughs was increased by inactive soy flour because soy proteins became insoluble and precipitated together with GMP. Active soy flour decreased GMP content due to gluten depolymerization. CONCLUSION: Soy proteins were associated to wheat protein through physical interaction and covalent and non‐covalent bonds during mixing and resting. These interactions produced large and medium‐size polymers. This fact increased SDS solubility of insoluble gluten proteins, producing a weakening of the gluten network. Physicochemical status of soy protein in the product had a great influence on how wheat–soy proteins interact. Copyright © 2007 Society of Chemical Industry     

Relative dominance of physical versus chemical effects on the transport of adhesion-deficient bacteria in intact cores from South Oyster, Virginia

Bacterial transport experiments were conducted using intact sediment cores collected from sites on the Delmarva Peninsula near South Oyster, VA, to delineate the relative importance of physical and chemical heterogeneity in controlling transport of an adhesion-deficient bacterial strain. Electron microscopy revealed that the sediments consisted of quartz and feldspar with a variable amount of clay and iron and aluminum hydroxide coatings on the grains. A nonmotile, gram-negative indigenous groundwater strain, designated as Comamonas sp. DA001, was injected into the cores along with a conservative tracer bromide (Br). DA001 cells were 1.2 x 0.6 microm in size with a hydrophilic surface and a slightly negative surface charge. Bacterial breakthrough preceded that of Br. This differential advection phenomenon can be accounted for by reduction of the effective porosity for the bacteria relative to Br. The distribution of cells remaining in the core as determined by scintillation counting and phosphor imaging techniques was variable, ranging from nearly uniform concentrations throughout the core to exponentially decreasing concentrations with distance from the point of injection. The fraction of bacterial retention in the core was positively correlated with the abundance of the metal hydroxides and negatively correlated with grain size. Because grain size was inversely correlated with the abundance of the metal hydroxide coatings, it was necessary to separate the effects of grain size and mineralogy. The fraction of the bacterial retention accounting for the effect of grain size, the collision efficiency, exhibited no correlation with the abundance of the metal hydroxides, indicating that the bacterial retention was primarily controlled by grain size. Reasons for the lack of influence of mineralogy on bacterial transport include (i) the slightly negatively charged bacterial surfaces; (ii) an insufficient heterogeneity of sediment surface properties; and (iii) the masking of the positive charge of the metal hydroxide surfaces by adsorbed organic carbon (up to 1180 ppm). This study demonstrates that the laboratory-based bacterial transport experiments are effective in delineating physical versus chemical controlling factors and provide an important link to field-based bacterial transport studies.