Oxygen transfer into a medium with extracellular polysaccharide

Measurement of oxygen transfer into a pseudoplastic solution of agrobacteran yielded the correlationK _L ^a ≈Re ^1.013 (100Re 600). A possible method of estimation of changes inK _L ^a, in dependence on agrobacteran concentration is presented.     

Cellulose as extracellular polysaccharide of hot spring sulfur-turf bacterial mat

The carbohydrate fraction of a hot spring sulfur-turf bacterial mat was shown to contain cellulose by the examination of neutral sugar composition, methylation analysis, and the identification of free oligosacchrides obtained from an acetolyzate of the desulfurized sulfur-turf mat. This suggested that the sulfur-oxidizing bacteria composing the sulfur-turf were producers of cellulose.     

Isolation and characterization of a new extracellular polysaccharide from an Acetobacter species

An expolysaccharide (EPS)-producing strain of the genus Acetobacter (named IFR 101) was isolated from a commercial vinegar acidifier. IFR 101 can be grown over the pH range 3.0–8.0 and the temperature range 12–35°C, and is ethanol-tolerant to 10% v/v. Neutral sugar analysis of the purified EPS indicates the presence of the sugars mannose, galactose and glucose. Methylation studies and NMR data suggest that the EPS may either be a complex branched polysaccharide or a mixture of simpler linear and branched structures. Aqueous preparations of IFR 101 exhibit reversible shear-thinning behaviour.     

Study on optimization of moisture retention for golf green rootzone soil mixtures

Most golf course green have been constructed with pure sand or sand-based rootzone mixes. As we know, high sand content provides rapid drainage despite sand’s inefficiency in retaining moisture. However, drainage capability and water retention are both essential elements to the golf course green, and the addition of peat could increase the soil moisture retention, therefore, the research on the drainage capability and water retention of the sandy golf green has become more and more important these years. In this study, extreme vertex design was applied which is one of the mixture experiment designs widely used in mixture experiments, the study investigated the effects of the thirteen different rootzone soil mixtures using middle-coarse, fine sand, very fine sand plus silt and clay as well as peat as the materials under three kinds of golf green profile (1-layer profile, 2-layer profile, 3-layer profile) conditions on the water retention of green rootzone. Through the qualitative, quantitative and optimization analysis of water retention capability of the sandy golf green, evidence a basis for choice of green profiles and rootzone matrix could be provided. And the significantly correlative regression model was established between the moisture retention and components of rootzone soil mixture. In addition, the order of factor contribution ratio, effect of single and double factor and optimization of the model were analyzed in detail. The results were as follows: both green profile and soil mixture, which had interaction of each other, had significant effects on soil moisture retention. Additional attributes include high porosity and greater water holding capacity than sand, and the higher content of peat, fine sand plus silt and clay, the better water retention. The mixtures had much higher water content in 1-layer profile than that in the other two profiles. There was significantly higher water content in 2-layer profile for pure sand mixtures (A–E) and low peat mixture (F) than that in 3-layer profile, while there was no higher water content for other 7 high peat mixtures (G–M) (>5%) in 2-layer than that in 3-layer profiles. The significance of key factors in rootzone soil mixture on moisture retention were: very fine sand plus silt and clay > peat > middle-coarse > fine sand. According to the moisture retention 15–25% specification of USGA (United States Golf Association), the optimal soil mixture in 1-layer profile was: middle-coarse 71.4–73.5%; fine sand 17.8–21.5%; very fine sand plus silt and clay 6.8–8.4%; peat 0–1%. The optimal soil mixture in 2-layer profile was: middle-coarse 65.0–73.4%; fine sand 17.8–20.5%; very fine sand plus silt and clay 7.5–8.9%; peat 0.2–6.3%. The optimal soil mixture in 3-layer profile was: middle-coarse 62.3– 73.9%; fine sand 17.7–21.4%; very fine sand plus silt and clay 7.3–10.7%; peat 0–6.3%. These optimal recipes took through the limitation of previous research, which were practically important to golf green soil selection and profile design. Thus, both proportion and interaction should be considered when we choose the soil mixture.     

灰树花胞外多糖的结构及免疫调节活性

对灰树花胞外多糖A组分(EXGFP-A)进行结构分析和免疫活性的研究。结构分析结果表明,EXGFP-A是一种主要含有葡萄糖的吡喃型中性多糖。气相结果说明EXGFP-A的单糖组成为鼠李糖、阿拉伯糖、木糖、甘露糖、葡萄糖、半乳糖,摩尔比为0.28∶0.31∶0.30∶0.06∶7.98∶0.61。MTT(3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazo-lium bromide)比色实验表明,当EXGFP-A浓度为80μg/m L,作用48 h时,RAW264.7细胞增殖指数达到最大值,为137.5%。吖啶橙(AO)染色结果表明EXGFP-A能够激活RAW264.7细胞,增强细胞内部核酸代谢水平。EXGFP-A在一定浓度范围内,可以提高RAW264.7细胞中NO的释放量,上调细胞内TNF-α、IL-1β、IL-6、IL-12、IFN-γ细胞因子以及细胞中iNOS的mRNA水平的表达。结果表明,EXGFP-A具有一定的免疫调节活性,为灰树花胞外多糖的结构分析和应用提供了科学依据。     

Immune-stimulating activity of water-soluble extracellular polysaccharide isolated from Rhizobium massiliae

The aim of this study is to investigate biochemical properties of water-soluble extracellular polysaccharide (WSP) from a novel bacterial strain designated to CA-1 and classified to Rhizobium massiliae by 16S rDNA sequence determination and homology analysis. The main composition of WSP was determined to be glucose by HPAEC. We evaluated immunomodulatory effects of WSP on RAW 264.7 macrophage activation. The results showed that the WSP dose-dependently induced the release of the pro-inflammatory cytokines such as TNF-α and IL-6. In addition, WSP induced nitric oxide synthase (iNOS) expression and increased the production of nitric oxide (NO). Intriguingly, WSP remarkably increased the mRNA expression of Toll-like receptor- 2 (TLR-2) and the phosphorylation of MAPKs (ERK, JNK and p38) in RAW 264.7 cells. These results indicated that WSP activates macrophages to secrete pro-inflammatory cytokines and induces iNOS expression via the activation of the TLR-2/MAPKs signaling pathways. Conclusively, we suggest that WSP of R. massiliae CA-1 can be a new immunomodulatory enhancing the early innate immunity.     

Fluorescence anisotropy as a means to determine extracellular polysaccharide hydrolase activity in environmental samples

Current approaches to measure the activities of microbial extracellular enzymes in aquatic environments are hampered by slow throughput or by differences between the structure of simple substrate proxies and macromolecules. Here we show that measurements of fluorescence anisotropy can be used to determine the hydrolysis rate of two fluorescently labeled polysaccharides, laminarin and xylan, in environmental samples. A simple analysis shows that the anisotropy of these fluorescently labeled polysaccharides can be approximated using a modification of the Perrin equation.     

Selection of a chemically defined medium for submerged cultivation of Streptomyces aureofaciens with high extracellular caseinolytic activity

A chemically defined medium was developed for the submerged cultivation of Streptomyces aureofaciens with a high secretion of caseinolytic activity. The medium composition is: 40 g/liter maltose; 1.640 g/liter L-leucine (0.0125M); 1.765 g/liter L-lysine (0.0125M); 6.976 g/liter K2HPO4 (0.04M); 4 g/liter CaCO3; 0.2 g/liter MgSO4.7H2O; 0.01 g/liter ZnSO4.7H2O; 0.01 g/liter FeSO4.7H2O: 0.01 g/liter MnSO4H2O, and 0.005 g/liter CoSO4.7H2O. Quantitative correlations were established between the concentrations of nutrients in the medium and the secretion of proteolytic activity. In this medium the secretion of proteolytic activity parallels growth, reaching a maximum after 70 hr at 30 degrees C in shaker cultures. The secretion appears to be an active process and to require aerobic conditions.     

产弹性蛋白酶菌种的筛选及发酵条件优化研究

从合肥肉联厂附近的土壤和污水中分离得到19株产弹性蛋白酶菌株,初步鉴定该菌株属于假单胞菌属.经过发酵复筛有三株产酶能力超过15u/mL.实验对菌株最佳产酶发酵条件进行了优化2%干酪素、0.5%葡萄糖、0.4%酵母膏、0.2% K2HPO4、0.01% MgSO4·7H2O;起始pH值7.0;最适发酵温度为30℃;装液量为25mL/250mL;该菌株在28h左右产弹性蛋白酶的量达18u/mL.     

Enhanced antibiotic activity of Xenorhabdus nematophila by medium optimization

Nutrition had highly influence on the antibiotic production by Xenorhabdus nematophila YL001. Glucose and peptone were identified as the best carbon and nitrogen sources that significantly affected antibiotic production using one-factor-at-a-time approach. Response surface methodology was applied to optimize the medium constituents (Glucose, peptone and minerals) for antibiotic production by X. nematophila YL001. Higher antibiotic activity (328.9 U/ml) was obtained after optimizing medium components. The optimal levels of medium components were (g/l): glucose 6.13, peptone 21.29, MgSO(4).7H(2)O 1.50, (NH(4))(2)SO(4) 2.46, KH(2)PO(4) 0.86, K(2)HPO(4) 1.11 and Na(2)SO(4) 1.72. An overall 16% and 35% increase in antibiotic activity were obtained as compared with mean observed response (283.7U/ml) at zero level of all variables and YSG medium.     

Enzymatic synthesis of a catecholic polyphenol product with excellent antioxidant activity

Abstract

Polyphenols, especially catecholic stilbene derivatives, have attracted much attention due to the huge pharmacological effects and promising health benefits. However, their chemical synthesis via regioselective ortho-hydroxylation on aromatic rings is highly challenging. In this study, 3′-hydroxypterostilbene (HPS) is taken as a model product due to its strong potential as an antitumor agent. One-step enzymatic synthesis of HPS from pterostilbene (PS) was explored, with immobilised tyrosinase as catalyst. The impact of solvent, pH, temperature, oxygen and reductant concentration on the reaction was investigated, and the conversion was optimised by employing the response surface methodology (RSM). Finally, a high yield of 77.9% was obtained in 2.7?h. This study demonstrates the first successful use of a biotechnological strategy to synthesise HPS. The antioxidant activities of both PS and HPS were evaluated by using the DPPH assay, demonstrating that HPS is more potent than PS as a radical scavenger.     

Structure and properties of a bacterial polysaccharide named Fucogel

The chemical structure of a polysaccharide named Fucogel was characterized and the position of acetylation was identified by NMR. A conformational analysis was performed on this 3-sugar repeating unit. From this, the persistence length, characterizing the stiffness of the polysaccharide, was determined and the role of the presence of acetyl group, reducing the stiffness, was pointed out. The helical conformations were also predicted, one of these being in agreement with X-ray data obtained on a similar polysaccharide. Experimental characterization of the native and deacetylated polysaccharides was developed. SEC experiments allowed us to determine the molar mass and the persistence length on the deacetylated polysaccharide. The value is in good agreement with that predicted from the molecular modeling. Microcalorimetry, rheology, and fluorescence spectroscopy demonstrated respectively that no helical conformation exists in solution but that loose interchain interactions due to the acetyl substituents exist in dilute solutions.     

Influence of the culture conditions on extracellular lyase activity related to K5 polysaccharide

The K5 capsular polysaccharide antigen of some Escherichia coli strains is the non-sulphated precursor in heparin biosynthesis. It is composed by two components, 16000 and 1500 Da, whose ratio depends on the activity of the extracellular form of a lyase synthesized by the same K5 producer strain. The lyase activity on the K5 chain size was greatly influenced by the medium composition employed for the lyase production. The control of lyase activity results in defined ratios of the two components in the K5 polysaccharide that is suitable for the semisynthetic production of heparin-like molecules.     

Activation of bacterial lytic polysaccharide monooxygenases with cellobiose dehydrogenase

Lytic polysaccharide monooxygenases (LPMOs) represent a recent addition to the carbohydrate‐active enzymes and are classified as auxiliary activity (AA) families 9, 10, 11, and 13. LPMOs are crucial for effective degradation of recalcitrant polysaccharides like cellulose or chitin. These enzymes are copper‐dependent and utilize a redox mechanism to cleave glycosidic bonds that is dependent on molecular oxygen and an external electron donor. The electrons can be provided by various sources, such as chemical compounds (e.g., ascorbate) or by enzymes (e.g., cellobiose dehydrogenases, CDHs, from fungi). Here, we demonstrate that a fungal CDH from Myriococcum thermophilum (MtCDH), can act as an electron donor for bacterial family AA10 LPMOs. We show that employing an enzyme as electron donor is advantageous since this enables a kinetically controlled supply of electrons to the LPMO. The rate of chitin oxidation by CBP21 was equal to that of cosubstrate (lactose) oxidation by MtCDH, verifying the usage of two electrons in the LPMO catalytic mechanism. Furthermore, since lactose oxidation correlates directly with the rate of LPMO catalysis, a method for indirect determination of LPMO activity is implicated. Finally, the one electron reduction of the CBP21 active site copper by MtCDH was determined to be substantially faster than chitin oxidation by the LPMO. Overall, MtCDH seems to be a universal electron donor for both bacterial and fungal LPMOs, indicating that their electron transfer mechanisms are similar.     

Alternative methods to limit extracellular bacterial activity for enumeration of intracellular bacteria

The gentamicin survival assay, a method routinely used to estimate bacterial infection of eukaryotic host cells, depends on the presumed limited penetration of gentamicin across the eukaryotic cell membrane. However, some studies have suggested that gentamicin may in fact enter eukaryotic cells and kill intracellular bacteria. In this study we devised alternative methods to enumerate intracellular Salmonellae using a lytic bacteriophage, SP6, and an amino acid auxotroph, Pro- mutant, which replicates selectively within host cells in the presence of its uptake inhibitor, 3,4-dehydro-L-proline. The conventional gentamicin survival assay was systematically compared with the alternative methods for the enumeration of intracellular Salmonellae. We found that gentamicin decreases the survival of intracellular Salmonellae when added to extracellular media at concentrations above 20 microg/ml. The alternative methods do not suffer from this disadvantage, suggesting that they should be used to replace the gentamicin survival assay. In addition, the proline auxotroph method could be applied to detect bacterial release from host cells.     

Polyelectroyte behavior of a bacterial polysaccharide from Xanthomonas campestris: Comparison with carboxymethylcellulose

The main physicochemical properties of the polysaccharide called Xanthan produced by Xanthomonas compestris are discussed: the activity coefficient of the counter-ion, the pK(α), and the ionic selectivity are investigated and compared to those of a carboxymetholcellulose. The weight-average molecular weight (M _w = 2 × 10⁶), the intrinsic viscosity and the constant of sedimentation are determined as a function of the ionic strength. It is proved that in dilute solution, there is no intermolecular association, whatever the ionic strength. The conformation is proposed to be a rigid rodlike molecule whose length is 6000 Å, independent of ionic strength > 10^?2N.     

A new method for the quantitative determination of microbial extracellular polysaccharide production using a disposable ultrafilter membrane unit

A new method was developed for the determination of extracellular polysaccharides produced by microorganisms. Culture supernatants were ultrafiltered through a membrane having a molecular exclusion limit of 5000 Da. Mono- and disaccharides were removed by repeated washings with water and then the polysaccharide in the retentate was determined by colorimetry. The method has been successfully applied to determine polysaccharide production by Lactobacillus delbrueckii subsp. bulgaricus NCFB 2483 in milk.     

Characterization of an extracellular polysaccharide from a xanthomonas species

An extracellular polysaccharide containing glucose, mannose, D-manno-octulosonic acid (KDO), an unidentified component (X), and acetyl groups in the molar ratio of 1.3:3.8:1.6:1.1:2.9, was obtained from the incubated medium of a Xanthomonas species. The extracellular polysaccharide contained traces of phosphate and nitrogen but no lipid. Mild hydrolysis with 0.025M sulfuric acid released all of the KDO in the polysaccharide and a KDO-free product was obtained, which on hydrolysis with 0.05M sulfuric acid, gave mainly an oligosaccharide containing glucose, mannose, and X in molar ratio of 1:1:1. The reducing end-group of this oligosaccharide was X, and other hexose residues were linked (1 → 4). Compound X seems to be a 6-deoxyhexose that differs from fucose and rhamnose.     

Production of an extracellular polysaccharide with emulsifier properties by Penicillium citrinum

Penicillium citrinum produced a glycolipid with emulsifier properties during cultivation on mineral medium with 1% (v/v) olive oil as carbon source. The emulsifier production was growth-associated and reached maximal activity at 60 h of cultivation. The production yield (Y _p/s) was 0.54 and the best emulsifying activity was observed for xylene and diesel oil when compared to other carbohydrates tested. The emulsifier was shown to be stable to a wide range of pH and temperature values and was shown to contain D-galactose, D-glucose and D-xylose (8.2:1.0:5.3) with a total carbohydrate content of 43%. The presence of salts stimulated the emulsification activity, suggesting potential for its application in industrial waste or marine remediation.