Structural and Rheological Properties of Exopolysaccharides Produced by Some Lactic Acid Bacterial Strains Isolated from Palm Wine

Aims: This study evaluated the physical, chemical and rheological properties of exopolysaccharides (EPSs) produced by Lactic Acid Bacteria (LAB) isolated from palm wine. Materials and Methods: EPSs from palm wine LAB strains were produced on 6% sucrose broth, purified and freeze-dried prior to analyses. Molecular weights (MW), rheological and structural composition (functional groups) of the EPSs were determined using standard methods and Fourier transform infrared spectroscopy (FTIR). Results: The average MW of the EPSs ranged from 2.02×106 to 6.53×106 Da while the flow index (n) values ranged from 0.03-3.13 at 0.2%, 0.06-1.51 at 0.4%, 0.38 - 1.85 at 0.6%, 0.14 - 2.26 at 0.8% and 0.55 - 6.42 at 1% concentrations at elevated temperatures for EPS solutions from the ten LAB species. The FTIR spectrum revealed prominent peaks of various groups of OH (3420 cm-1) and CH3 bending (2090 cm-1) in all the EPSs corresponding to both hydroxyl and amine groups, and aliphatic C-H bonds, respectively. EPS synthesized by Leuconostoc lactis, Lactococcus lactis subsp lactis and Lactiplantibacillus plantarum showed weak absorption peaks (1148 – 1145 cm-1) indicating the C-O-C and C-O bonds, while absorption peaks of Lactobacillus lactis, Lactobacillus acidophilus and Lactiplantibacillus plantarum (1267 – 1253 cm-1) indicated O- acetyl ester and other non-sugar components. Conclusions: The FTIR spectra, rheological properties and molecular weight of EPSs synthesized by the ten LAB strains indicated potentials that could be exploited in different industrial applications, and as stabilizers in food industries

Aislamiento y caracterización de bacterias productoras de biopolimeros a partir de efluentes industriales / Isolated and characterization of biopolymer producing bacteria from industrial effluents

RESUMEN Se realizó una caracterización físico-química de los efluentes procedentes de industrias del sector educación, metalmecánico, lácteos y confitería de la ciudad de Manizales, Caldas; posteriormente se obtuvieron aislamientos, en medios diferenciales suplementados, de bacterias con potencial para la producción de biopolímeros a los cuales se les aplicó pruebas para la caracterización morfológica, bioquímica y molecular. Los parámetros físico químicos obtenidos de los efluentes industriales demuestran diferencias entre ellos, ya que cada industria genera diferentes residuos aportando una determinada contaminación al efluente, se obtuvieron 73 aislamientos productores de exopolisacáridos (EPS) y 101 productores de polihidroxialcanoatos (PHA), con características morfológicas y bioquímicas variables. El estudio muestra que los efluentes industriales son una gran fuente de bacterias de interés para la producción de diversos polímeros microbianos; principalmente aquellos que producen polímeros tipo biopoliésteres intracelulares como PHA, debido a su variabilidad físico-química y nutricional permitiendo que los microorganismos se adapten a diversas características medioambientales y de composición.

ABSTRACT A physical-chemical characterization of effluents from industries in the education, metal-mechanic, dairy and confectionery sectors of the city of Manizales, Caldas; Later isolates were obtained, in differential media supplemented, from bacteria with potential for the production of biopolymers to which they were applied tests for morphological, biochemical and molecular characterization. The physical chemical parameters obtained from the industrial effluents show a difference between them, since each industry generates different waste contributing a certain contamination to the effluent, 73 isolates producing exopolysaccharides (EPS) and 101 producers of polyhydroxyalkanoates (PHA) were obtained, with morphological characteristics and variable biochemistry. The study shows that industrial effluents are a great source of bacteria of interest for the production of various microbial polymers; mainly those that produce polymers like intracellular biopolyesters such as PHA, due to their physical-chemical and nutritional variability allowing the microorganisms to adapt to diverse environmental and compositional characteristics.

In vitro evaluation of α-glucosidase inhibitor and antioxidant activity of Lactobacillus isolates and their antidiabetic potential

Aims@#This study aimed to evaluate antidiabetic potential of indigenous Lactobacillus isolates by measuring the ability of α-glucosidase inhibitory (AGI) and antioxidant activity. The mechanism of probiotics as antidiabetic can occur through the AGI and antioxidant activity of LAB, which is able to suppress oxidative stress that causes chronic inflammation and pancreatic β cell apoptosis, and then through the ability to produce exopolysaccharide (EPS) and short chain fatty acids (SCFA).@*Methodology and results@#MRS broth enriched with 10% glucose was selected as the growth medium for Lactobacillus. The growth medium was then centrifuged to obtain CFS and CFE was produced by extracting the medium with 96% ethanol as a solvent. The results showed that Lactobacillus pentosus MK42 had the highest AGI activity of 80.32 ± 2.20%. Antioxidant activity was not significantly different (P>0.05) among the tested Lactobacillus isolates. Lactobacillus paracasei RK41 produced the highest EPS (360.13 ± 50.01 mg/L), which was not significantly different (P>0.05) from Lactobacillus plantarum1 RB210. All Lactobacillus isolates were able to produce acetic acid, but not all were able to produce propionic and butyric acid. The highest propionic acid was produced by L. plantarum1 RB210 at 0.40 ± 0.31 mmol/L and the highest butyric acid was produced by L. plantarum1 MK2 at 0.22 ± 0.08 mmol/L.@*Conclusion, significance and impact of study@#The results show definitively that indigenous Lactobacillus isolates have considerable α-glucosidase inhibitor, antioxidant activity and the ability to produce of EPS and SCFA. This preliminary study suggests the use of indigenous Lactobacillus isolates which have the potential as antidiabetic agent, although the responsible compounds are unknown.

Additive effect of the probiotics Lactobacillus exopolysaccharides and the Satureja calamintha extracts on enteropathogenic Escherichia coli adhesion

Abstract This study assessed the inhibitory potential of the probiotics Lactobacillus (LB) exopolysaccharides (EPS) with or without extracts of Satureja calamintha on enteropathogenic Escherichia coli (EPEc) responsible for gastroenteritis. Methanolic and hydromethanolic extracts were prepared by cold maceration and subjected to phytochemical screening. The compounds of the extracts were determined with the colorimetric assays and identified using high-performance liquid chromatography coupled with diode array detector (HPLC-DAD). Antioxidant activities of the extracts were also evaluated by using 2,2-diphenyl-1-picrylhydrazil (DPPH) radical scavenging. Antibacterial effect on EPEc was evaluated by using both agar disc diffusion and microdilution methods. The in vitro test of auto-aggregation was investigated. Microbiological analysis showed that 63% of the isolated LB were producing EPS, with the amount ranging from 8.21 to 43.13 mg/L. Chemical analysis of the extracts revealed the presence of polyphenols and flavonoids, more abundant in the hydromethanolic extract, which presented the highest content with 2.11 mg EGA/g of polyphenol and 1.64 mg EC/g of flavonoids and 1.71 mg EGA/g of polyphenol and 1.15 mg EC/g of flavonoids in the methanolic extract. Hydromethanolic extracts and EPS exhibited a more important activity than did the methanolic extract against EPEc. The combined action of EPS and extracts reduced the aggregation ability of EPEc and decreased the rate of their adhesion.

Potential Biotechnological Applications of Cyanobacterial Exopolysaccharides

Abstract The cyanobacterial exopolysaccharides (EPSs) are considered as one of the important group of biopolymers having significant ecological, industrial, and biotechnological importance. Cyanobacteria are regarded as a very abundant source of structurally diverse, high molecular weight polysaccharides having variable composition and roles according to the organisms and the environmental conditions in which they are produced. Due to their structural complexity, versatility and valuable biological properties, they are now emerging as high-value compounds. They are possessing exceptional properties and thus are being widely explored for various applications like in food and pharmaceutical industries, in bioremediation for removal of heavy metals, for soil conditioning, as biopolymers, bioadhesives, and bioflocculants. However, poor understanding of their complex structural properties, lack of concrete information regarding the genes encoding the proteins involved in the EPS biosynthetic pathways, their process of production and about the associated factors controlling their structural stability, strongly limits their commercialization and applications in the various fields of biotechnology. Owing to the above context, the present review is aimed to organize the available information on applications of cyanobacterial EPSs in the field of biotechnology and to identify the research gaps for improved industrial utilization and commercialization of these biomaterials.

Production of antioxidative exopolysaccharides of Cordyceps militaris with Vernonia amygdalina leaves in substrate / 生物工程学报

Cordyceps militaris exopolysaccharides (EPS) have many pharmacological activities such as boosting immunity and antifatigue. To obtain EPS efficiently, we added moderate Vernonia amygdalina leaf powder as inducer to the fermentation medium to promote the production of Cordyceps militaris EPS and studied the infrared absorption spectrum and antioxidant activities of the EPS after optimization. The optimum liquid fermentation conditions were as follows: addition of Vernonia amygdalina leaf powder of 8 g/L, fermentation duration of 9 d, initial pH of 6.5, inoculation quantity of 5.0 mL. Under such a condition, the yield of Cordyceps militaris EPS reached (5.24±0.28) mg/mL, increased by 205.20% compared to the control group without adding Vernonia amygdalina leaf powder. Results of infrared analysis and antioxidant activity showed that the Vernonia amygdalina leaves had little effect on the structure and activities of Cordyceps militaris EPS. The results of this research suggest that Vernonia amygdalina leaf can enhance the production of Cordyceps militaris EPS effectively, and provides a novel method for efficient production of EPS in Cordyceps militaris.

Research Methods for Structural Analysis of Lactic Acid Bacteria Induced Exopolysaccharides / 分析化学

Exopolysaccharides (EPSs), produced by lactic acid bacteria (LAB), have been used primarily to improve the quality and taste of food, also possess a variety of unique biological functions, such as immunoregulation and anti-tumor activities. The diversity of molecular structural characteristics of LAB-generated EPSs represents one of the main factors responsible for this plethora of functions. Accordingly, the structural analysis of the EPSs produced by LAB is both a prerequisite and basis for examining its functional and structure-activity relationships. In this article, we summarized the current progress of key methodologies involved in the structural analysis of LAB-generated EPSs, including their isolation, purification, primary structure and advances in structural research. A comprehensive discussion regarding the application of chemical analysis, instrument analysis and computer aided technology in the structure analysis of LAB-generated EPSs was provided. Further, the future development of the research on the structure of LAB-generated EPSs was presented.

Function of sahH gene in exopolysaccharides synthesis of Streptococcus mutans LuxS null strain / 上海交通大学学报（医学版）

Objective·To investigate the function of sahH gene in exopolysaccharides synthesis of Streptococcus mutans LuxS null strain. Methods·sahH+luxS-SmUA159 was constructed by introducing sahH gene into Streptococcus mutans LuxS null strain,while PIB169+luxS-SmUA159 and its wild strain SmUA159 were used as vector control and blank control. Real-time qPCR and anthrone test were performed to investigate the differences in exopolysaccharides synthesis among sahH+luxS-SmUA159, PIB169+luxS-SmUA159 and the wide-type strain SmUA159. Results·The transcriptional levels of gtfA and gtfD were upregulated in the sahH+luxS-SmUA159,compared to SmUA159 and PIB169+luxS-SmUA159(both P<0.05),but there was no statistical difference in the transcriptional levels of gtfB and gtfC.The results of the anthrone test showed exopolysaccharides synthesis of the three groups was also similar. Conclusion·sahH gene affects the expression of some genes related to exopolysaccharides synthesis,but it has little effect on the whole content of bacterial exopolysaccharides.

Function of sahH gene in exopolysaccharides synthesis of Streptococcus mutans LuxS null strain / 上海交通大学学报（医学版）

Objective: To investigate the function of sahH gene in exopolysaccharides synthesis of Streptococcus mutans LuxS null strain. Methods: sahH+luxS-SmUA159 was constructed by introducing sahH gene into Streptococcus mutans LuxS null strain, while PIB169+luxS-SmUA159 and its wild strain SmUA159 were used as vector control and blank control. Real-time qPCR and anthrone test were performed to investigate the differences in exopolysaccharides synthesis among sahH+luxS-SmUA159, PIB169+luxS-SmUA159 and the wide-type strain SmUA159. Results: The transcriptional levels of gtfA and gtfD were upregulated in the sahH+luxS-SmUA159, compared to SmUA159 and PIB169+luxS-SmUA159 (both P<0.05), but there was no statistical difference in the transcriptional levels of gtfB and gtfC. The results of the anthrone test showed exopolysaccharides synthesis of the three groups was also similar. Conclusion: sahH gene affects the expression of some genes related to exopolysaccharides synthesis, but it has little effect on the whole content of bacterial exopolysaccharides.

The exopolysaccharides biosynthesis by Candida yeast depends on carbon sources

Background: The exopolysaccharides (EPS) produced by yeast exhibit physico-chemical and rheological properties, which are useful in the production of food and in the cosmetic and pharmaceutical industries as well. The effect was investigated of selected carbon sources on the biosynthesis of EPS by Candida famata and Candida guilliermondii strains originally isolated from kefirs. Results: The biomass yields were dependent on carbon source (sucrose, maltose, lactose, glycerol, sorbitol) and ranged from 4.13 to 7.15 g/L. The highest biomass yield was reported for C. guilliermondii after cultivation on maltose. The maximum specific productivity of EPS during cultivation on maltose was 0.505 and 0.321 for C. guilliermondii and C. famata, respectively. The highest EPS yield was found for C. guilliermondii strain. The EPS produced under these conditions contained 65.4% and 61.5% carbohydrates, respectively. The specific growth rate (µ) of C. famata in medium containing EPS as a sole carbon source was 0.0068 h-1 and 0.0138 h-1 for C. guilliermondii strain. Conclusions: The most preferred carbon source in the synthesis of EPS for both Candida strains was maltose, wherein C. guilliermondii strain showed the higher yield of EPS biosynthesis. The carbon source affected the chemical composition of the resulting EPS and the contribution of carbohydrate in the precipitated preparation of polymers was higher during supplementation of maltose as compared to sucrose. It was also found that the EPS can be a source of carbon for the producing strains.

Small scale production and characterization of xanthan gum synthesized by local isolates of Xanthomonas campestris.

Xanthan gum is a commercially important microbial exopolysaccharide (EPS) produced by Xanthomonas campestris. X. campestris is a plant pathogen causing various plant diseases such as black rot of crucifers, bacterial leaf blight and citrus canker disease resulting in crop damage. In this study, we isolated efficient local bacterial isolates which are capable to produce xanthan gum utilizing different sources of carbon (maltose, sucrose and glucose). Bacterial isolates from different plant leaves and fruits were identified as Xanthomonas campestris based on their morphological and biochemical characteristics. Among the 23 isolates, 70% were capable of producing gum. Taro plant, considered as new bacterial host, also have the capability to produce xanthan gum. Production conditions of xanthan gum and their relative viscosity by these bacterial isolates were optimized using basal medium containing commercial carbon and nitrogen sources and various temperature and rotation. Highest level of xanthan gum (18.286 g/l) with relative viscosity (7.2) was produced (Host, Citrus macroptera) at 28°C, pH 7.0, 150 rpm using sucrose as a carbon source at orbital shaker. Whereas, in lab fermenter, same conditions gave best result (19.587 g/l gum) with 7.8 relative viscosity. Chilled alcohol (96%) was used to recover the xanthan gum. FTIR studies also carried out for further confirmation of compatibility by detecting the chemical groups.

Plant Growth Promotion and Root Colonization by EPS Producing Enterobacter sp. RZS5 under Heavy Metal Contaminated Soil.

The heavy metal resistant bacterium isolated from field soil and identified as Enterobacter sp. RZS5 tolerates a high concentration (100-2000 mM) of various heavy metal ions such as Mn2+, Ni2+, Zn2+, Cu2+, CO2+ and Fe2+ when grown in such environment and produces exopolysaccharides (EPS). Here, we have demonstrated EPS production by Enterobacter sp. RZS5 during 60 h of growth in yeast extract mannitol broth (YEMB). The yield increased by two fold after the addition of 60 M of Ca2+; 50 M of Fe2+ and 60 M of Mg2+ ions in YEMB, and the optimization of physico-chemical parameters. EPS was extracted with 30% (v/v) of isopropanol as against the commonly used 50% (v/v) isopropanol method. EPS-rich broth promoted seed germination, shoot height, root length, number of leaves and chlorophyll content of wheat (Triticum aestivum) and peanut (Arachis hypogaea) seeds. The higher colony-forming unit of Enterobacter sp. in soil inoculated with EPS rich broth of Enterobacter sp. indicated the root colonizing potential and rhizosphere competence of the isolate. The FTIR spectra of the EPS extract confirmed the presence of the functional group characteristics of EPS known to exhibit a high binding affinity towards certain metal ions. This overall growth and vigour in plants along with the effective root colonization, reflected the potential of the isolate as an efficient bio-inoculant in bioremediation.

Microbial Exopolysaccharides: Natural Modulators of Dairy Products.

Microbial exopolysaccharides (EPS) are considered as natural bio-thickeners abundantly used in dairy and fermented food industries for quality improvement. In fermentation based dairy industries, researchers are seeking their attention on substitution of artificial food stabilizers with naturals by exploring EPS, especially for improving the rheology of fermented food products. Lactic acid bacteria (LAB) being a starter player for fermented food products are believed to be as one of the best natural producers of EPS. These EPS are basically sugar residues, secreted by microbes in their surrounding environment but their in-vitro production is not economical due to the media and processing costs involved. The process of production is also complex due to the involvement of various enzymes and regulatory proteins. With modernization in dairy sector, our understanding towards EPS needs improvement. The review focuses on a brief explanation on the behavior and functionality of EPS from lactic acid bacteria.

Optimization of cultural conditions for exopolysaccharides production by Frateuria aurentia.

The exopolysaccharides (EPS) producing ability was evaluated by nutritional defined media by the Frateuria aurentia was the highest yield EPS-producing strain, which was isolated from elephant dung in Srivilliputhur, Tamilnadu, India. It was identified as Frateuria aurentia with carbohydrate assimilation process and 16S rDNA gene sequencing. The favourable conditions for EPS yield by the microorganism growth in a chemically defined medium were tested with carbon, nitrogen, cultivation time, temperature, pH, hydrocarbon, surfactants, sodium chloride, metal ions and inoculums concentration in static and shaking conditions had the most significant influences. Among the various nutritional sources of interactive effects the maximum EPS production was recorded at pH 7.00 (1.431 ± 0.0096 OD); 30 C (1.661 ± 0.028 OD); Jaggery (1.185 ± 0.003 OD); ⁰ Tryptone (1.248 ± 0.011 OD); Ferric chloride (1.682 ± 0.022 OD); Glutamine (2.136 ± 0.067 OD); polyethyleneglycol (0.873 ± 0.041 OD); 2.5% NaCl (1.138 ± 0.023 OD); Neem oil (1.699 ± 0.176 OD); inoculum concentration 2.00% (1.625 ± 0.007 OD); The maximum EPS production was viewed at the 72hrs of incubation at static condition (1.58 ± 0.122 OD).

Optimization of exopolysaccharides production from a novel strain of Ganoderma lucidum CAU5501 in submerged culture

This study aimed at optimizing the medium of a new Ganoderma lucidum strain CAU5501 to enhance the yield of exopolysaccharides (EPS) and mycelial growth. Firstly, the suitable level of glucose, magnesium, phosphate and C/N ratio was determined by single factor experiment. Subsequently, the optimum concentrations of these medium components were investigated using the orthogonal matrix method. The results indicated that the higher levels of EPS were correlated with the level of cell growth when glucose concentration was studied (data no show). The optimum medium for EPS yield was found to be 70 g/l glucose, 5 C/N ratio, 2.5 g/l KH2PO4, 0.75 g/l MgSO4·7H2O, and for mycelial growth was 50 g/l glucose, 5 C/N ratio, 1.5 g/l KH2PO4, 0.5 g/l MgSO4·7H2O. When cultivated in the obtained optimal media in 3 L shake flask, compared to the basal medium, the EPS yield increased markedly from 1.003 to 1.723 g/l, and the mycelium formation was also markedly improved from 2.028 to 7.235 g/l. Results obtained in this study are beneficial to further study for enhancing the production of Ganoderma lucidum polysaccharides in large scale commercialized production.

Effect of cyanobacterial exopolysaccharides on salt stress alleviation and seed germination.

Effect of exopolysaccharides (EPS) produced by a consortium of cyanobacteria on germination of three crops wheat, maize and rice was studied at different salt concentrations. Production of EPS was found to be stimulated by salts, which in turn had a significant Na+ removal capability from aqueous solution. Seed germination, vigor index and mobilization efficiency in all the three crops remarkably improved when cyanobacterial EPS was applied. While germination improved significantly by 13 to 30%, mobilization efficiency increased marginally by 1.03 to 1.1 times and vigor index increased by 1.15 to 2.4 times in these crops in response to EPS under non-saline conditions. Salinity had an inhibitory effect on seed germination of all the species showing 18 to 54% reduction. However, in the presence of EPS, the salt induced inhibition diminished to 13 to 18%. Inhibitory effect of salt on chlorophyll concentration, vigor index and mobilization efficiency of the seedlings was much less in these crops in the presence of EPS, indicating the latter’s role in salt stress alleviation.

Production of biopolymers by Pseudomonas aeruginosa isolated from marine source

Two bacterial strains, Pseudomonas aeruginosa CMG607w and CMG1421 produce commercially important biopolymers. CMG607w isolated from the sediments of Lyari outfall to Arabian Sea synthesize the mcl-polyhydroxyalkanoates from various carbon sources. The production of PHAs was directly proportional to the incubation periods. Other strain CMG1421, a dry soil isolate, produced high viscous water absorbing extracellular acidic polysaccharide when it was grown aerobically in the minimal medium containing glucose or fructose or sucrose as sole source of carbon. The biopolymer had the ability to absorb water 400 times more than its dry weight. This property was superior to that of currently used non-degradable synthetic water absorbents. It acted as salt filter and had rheological and stabilizing activity as well.

Response of the Higher Basidiomycetic Ganoderma resinaceum to Sodium Chloride Stress

Ganoderma resinaceum tolerated sodium chloride salt stress within a range of 0 mM till 300 mM. It responded to salt stress with fluctuation in proline formation at different NaCl concentrations. However,the mycelial dry weight,total protein contents and exopolysaccharides did not changed considerably. Increasing sodium chloride concentration led to morphological alteration in fungal mycelia with disappearance of fungal cell wall,plasmolysis,and vacuolation as indicated with electron microscopic examination of the fungal growth.

Pleiotropic effects of purine auxotrophy in Rhizobium meliloti on cell surface molecules.

Rhizobial purine auxotrophs have earlier been shown to be defective in symbiosis, though the exact reason for this failure is not clear. Using various dyes that specifically bind different cell surface molecules, we show that there are multiple changes in the cell surface molecules associated with different purine auxotrophs. Affected molecules in different purine auxotrophs that were tested include (i) acidic exopolysaccharides, (ii) cellulose fibrils, and (iii) beta (1–3) glucans. Our results show that the symbiotic deficiency of purine auxotrophs is likely to be a result of these associated changes on the cell surface.

THE EFFECTS OF EXOPOLYSACCHARIDES FROM APHANOTHECE HALOPHYTICA (EAH ) ON IMMUNO-LOGICAL FUNCTION OF MICE / 中国海洋药物

The effects of exopolysaccharides from Aphanothece halophytica(.EAH) on immunological function were studied by immunological techniques. The results showed that EAH could not only enhance the mitogenic response of mice's thymus cells, murine bone marrow cells proliferation,murine mixed lymphocyte reaction and the activity of NK cells in mice,but promote the interleukinl(IL-1)production and increase the activity of 1L-2 as well. The study suggests that EAH can improve the immunological function of mice.