Cordyceps militaris, an entomopathogenic fungus belonging to the class Ascomycetes, has been reported to have beneficial biological activities such as hypoglycemic, anti-inflammatory, antitumor, antimetastatic, hypolipidemic, immunomodulatory, and antioxidant effects. In this study, the crude water-soluble polysaccharide CMP, which was obtained from the fruiting body of C. militaris by hot water extraction and ethanol precipitation, was fractionated by DEAE-cellulose and Sepharose CL-6B column chromatographies. This process resulted in three polysaccharide fractions, termed CMP Fr I, CMP Fr II, and CMP Fr III. Of these fractions, CMP Fr II, with an average molecular mass of 127 kDa, was able to upregulate effectively the phenotypic functions of macrophages such as NO production and cytokine expression. The chemical property of the stimulatory polysaccharide, CMP Fr II, was determined based on its monosaccharide composition, which consisted of glucose (56.4%), galactose (26.4%), and mannose (17.2%). Its structural characteristics were investigated by a combination of chemical and instrumental analyses, including methylation, reductive cleavage, acetylation, Fourier transform infrared spectroscopy (FTIR), and gas chromatography-mass spectrometry (GCMS). Results indicated that CMP Fr II consisted of the (1${\rightarrow}$4) or (1${\rightarrow}$2) linked glucopyranosyl or galactopyranosyl residue with a (1${\rightarrow}$2) or (1${\rightarrow}$6) linked mannopyranosyl, glucopyranosyl, or galactopyranosyl residue as a side chain. The configuration of the ${\beta}$-linkage and random coil conformation of CMP Fr II were confirmed using a Fungi-Fluor kit and Congo red reagent, respectively.

The biological synthesis of nanoparticles has gained considerable attention in view of their excellent biocompatibility and low toxicity. We isolated and purified rhamnolipids from Pseudomonas aeruginosa strain BS-161R, and these purified rhamnolipids were used to synthesize silver nanoparticles. The purified rhamnolipids were further characterized and the structure was elucidated based on one- and two-dimensional $^1H$ and $^{13}C$ NMR, FT-IR, and HR-MS spectral data. Purified rhamnolipids in a pseudoternary system of n-heptane and water system along with n-butanol as a cosurfactant were added to the aqueous solutions of silver nitrate and sodium borohydride to form reverse micelles. When these micelles were mixed, they resulted in the rapid formation of silver nanoparticles. The synthesized nanoparticles were characterized by UV-Visible spectroscopy, transmission electron microscopy, and energy dispersive X-ray spectroscopy (EDS). The nanoparticles formed had a sharp adsorption peak at 410 nm, which is characteristic of surface plasmon resonance of the silver nanoparticles. The nanoparticles were monodispersed, with an average particle size of 15.1 nm (${\sigma}={\pm}5.82$ nm), and spherical in shape. The EDS analysis revealed the presence of elemental silver signal in the synthesized nanoparticles. The formed silver nanoparticles exhibited good antibiotic activity against both Grampositive and Gram-negative pathogens and Candida albicans, suggesting their broad-spectrum antimicrobial activity.

A novel laccase from Tricholoma mongolicum was purified by using a procedure that entailed ion-exchange chromatographies on DEAE-cellulose, CM-cellulose, and Q-Sepharose, and FPLC-gel filtration on Superdex 75. The purified enzyme was obtained with a specific activity of 1,480 U/mg-protein and a final yield of 15%. It was found to be a monomeric protein with a molecular mass of 66 kDa as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Its N-terminal amino acid sequence was GIGPVADLYVGNRIL, similar to some but also different to other mushroom laccases. The optimum pH and temperature for the purified enzyme were pH 2 to pH 3 and $30^{\circ}C$, respectively. It displayed a low $K_m$ toward 2,7-azinobis (3-ethylbenzothiazolone-6-sulfonic acid) diammonium salt (ABTS) and high $k_{cat}/K_m$ values. The purified laccase oxidized a wide range of lignin-related phenols, but exerted maximal activity on ABTS. It was significantly inhibited by $Hg^{2+}$ ions, and remarkably stimulated by $Cu^{2+}$ ions. It inhibited HIV-1 reverse transcriptase and proliferation of hepatoma HepG2 cells and breast cancer MCF7 cells with an $IC_{50}$ of 0.65 ${\mu}M$, 1.4 ${\mu}M$, and 4.2 ${\mu}M$, respectively, indicating that it is also an antipathogenic protein.

With an emphasis on its thermal behavior with different pHs and salts, the kinetic and thermodynamic parameters of the purified polygalacturonase (PG) from E. carotovora subsp. carotovora (Ecc) BR1 were studied, as the characterization of an enzyme is significant in the context of burgeoning biotechnological applications. The thermodynamic parameters for polygalacturonic acid hydrolysis by the purified PG were ${\Delta}H^*$=7.98 kJ/mol, ${\Delta}G^*$=68.86 kJ/mol, ${\Delta}S^*$=-194.48 J/mol/K, ${\Delta}G_{E-S}$=-1.04 kJ/mol, and ${\Delta}G_{E-T}$=-8.96 kJ/mol. In addition, its turnover number ($k_{cat}$) was 21/sec. The purified PG was stable within a temperature range of $20-50^{\circ}C$ and was deactivated at $60^{\circ}C$ and $70^{\circ}C$. The thermodynamic parameters (${\Delta}H^*$, ${\Delta}G^*$, ${\Delta}S^*$) for the irreversible inactivation of the PG at different temperatures ($30-60^{\circ}C$) were determined, where the effectiveness of various salts and different pHs (4-8) for the thermal stability of the PG were also characterized. The efficacy of various salts for the thermal stability of the PG was in the following order: $MgCl_2$ > $BaCl_2$ > KCl > $CaCl_2$ >NaCl. Therefore, the present work presents the biochemical, substrate hydrolysis thermodynamics and the thermal stabilization parameters of the PG from Ecc.

This study explored the use of gellan gum as an immobilization matrix for the production of cyclosporin A (CyA) by immobilized spores and mycelia of Tolypocladium inflatum MTCC 557. Different carriers, such as gellan gum, sodium alginate, celite beads, and silica, were tested as immobilization carriers, along with the role of the carrier concentration, biomass weight, number of spore-inoculated beads, and repeated utilization of the immobilized fungus. The maximum CyA production was 274 mg/l when using gellan gum [1% (w/v)], and a mycelial weight of 7.5% (w/v) supported the maximum production of CyA. Additionally, the addition of a combination of $_L$-valine (6 g/l) and $_L$-leucine (5 g/l) after 48 h of fermentation produced 1,338 mg/l of CyA when using gellan gum. The immobilized mycelia beads were found to remain stable for four repetitive cycles, indicating their potential for semicontinuous CyA production.

Bacteria, fungi, and protozoa inhabiting the rumen, the largest chamber of the ruminants' stomach, release large quantities of hydrogen during the fermentation of carbohydrates. The hydrogen is used by coexisting methanogens to produce methane in energy-yielding processes. This work shows, for the first time, a fundamental possibility of using a hydrogen-rich fermentation gas produced by selected rumen ciliates to feed a low-temperature hydrogen fuel cell. A biohydrogen fuel cell (BHFC) was constructed consisting of (i) a bioreactor, in which a hydrogen-rich gas was produced from glucose by rumen ciliates, mainly of the Isotrichidae family, deprived of intra- and extracellular bacteria, methanogens, and fungi; and (ii) a chemical fuel cell of the polymer-electrolyte type (PEFC). The fuel cell was used as a tester of the technical applicability of the fermentation gas produced by the rumen ciliates for power generation. The average estimated hydrogen yield was ca. 1.15 mol $H_2$ per mole of fermented glucose. The BHFC performance was equal to the performance of the PEFC running on pure hydrogen. No fuel cell poisoning effects were detected. A maximum power density of $1.66\;kW/m^2$ (PEFC geometric area) was obtained at room temperature. The maximum volumetric power density was $128\;W/m^3$ but the coulombic efficiency was only ca. 3.8%. The configuration of the bioreactor limited the continuous operation time of this BHFC to ca. 14 h.

Pulsed electric field (PEF) of 1Hz, 1.5 kV, and 1ms increased the activities of catalase and inulinase over the whole range of applied Se concentrations compared with the non-treated cultures. A significant effect of selenium concentration (in the range of 5-14 ${\mu}g/ml$) on both intra- and extracellular enzyme activities was noted. At a Se concentration of 10 ${\mu}g/ml$, the activities of intra- and extracellular inulinases and extracellular catalase in the PEF-treated cultures reached the maximum of 71 U/g d.m., 46 U/g d.m., and approx. 8 U/ml, respectively. The maximum activity of intracellular catalase of approx. 6 U/ml (with and without PEF) was recorded at 5 ${\mu}g$ Se/ml. Further increasing of selenium concentration caused a decrease in the activity of the enzymes.

Because conventional methods for detecting emetic-toxin-producing B. cereus are laborious and costly, various PCR assays, which are easy and cheap, have recently been reported. Therefore, this study estimated and compared the ability of various PCR assays to detect emetic-toxin-producing B. cereus strains isolated in Korea. The PCR assays were performed on 160 B. cereus strains, including 40 emetic-toxin-producing strains. Although the species-specific PCR assays were all shown to be highly specific, the sensitivities varied greatly. The accuracies of the primers were 97.5% (CER), 95.6% (EM1), 96.3% (RE234), 89.4% (CES), and 83.1% (Ces3R/CESR2). Moreover, the CER primer had a higher sensitivity (100%) than all the other primers tested, and a specificity of 96.7%. Thus, the CER primer was shown to be the most effective for screening the emetic-toxin-producing B. cereus strains tested in this study. However, the ability of these PCR assays to identify emetic-toxin-producing B. cereus should also be confirmed using other methods.

A chitosan-degrading fungus, BSF114, was isolated from soil. The culture preparation showed strong chitosanolytic enzyme activity at an optimum pH of 4.0 and optimum temperature of $60^{\circ}C$ after 36-40 h fermentation. The rapid decrease in the viscosity of the chitosan solution early in the reaction suggested an endo-type cleavage of the polymeric chitosan chains. To identify the isolated fungus, molecular biological and morphological methods were used. The fungal internal transcribed spacer (ITS) region 1 was amplified, sequenced, and then compared with related sequences in the GenBank database using BLAST. The phylogenetic relationships were then analyzed, and the results showed that the fungus belongs to Aspergillus fumigatus. Morphological observations were also used to confirm the above conclusion. The chitooligosaccharides (COS) obtained through hydrolyzing the colloidal chitosan showed that A. fumigatus BSF114 is suitable for degrading chitosan and producing chitooligosaccharides on a large scale. High concentrations of the COS (1,000 and 500 ${\mu}g/ml$) significantly proliferated mice marrow cells.

This study was performed to investigate the effects of adding a dual starter on the chemical and sensory characteristics of red wine made of Campbell Early grape. The yeast starter, Saccharomyces cerevisiae, and lactic acid bacteria (LAB) starter, Oenococcus oeni, were used for inoculation in the winemaking process for alcoholic fermentation and malolactic fermentation (MLF), respectively. After 200 days of incubation, the chemical compositions of yeast/LAB-added wine (YL-wine) were compared with those of no-starter-added wine (control) and yeast-added wine (Y-wine). The results show that no significant differences were observed in pH, total sugar, and alcohol content among the wine samples, but the malic acid content in YL-wine was significantly reduced, and various esters and higher alcohols were synthesized. The sensory test revealed that the addition of dual starters resulted in improved overall acceptability in wine. This study emphasizes the importance of O. oeni in addition to yeast in making Campbell Early wine.

The diversity and abundance of ammonia-oxidizing bacteria (AOB) in activated sludge were compared using PCR-DGGE and real-time PCR assays. Activated sludge samples were collected from five different types of wastewater treatment plants (WWTPs) mainly treating textile, paper, food, and livestock wastewater or domestic sewage. The composition of total bacteria determined by PCR-DGGE was highly diverse between the samples, whereas the community of AOB was similar across all the investigated activated sludge. Total bacterial numbers and AOB numbers in the aerated mixed liquor were in the range of $1.8{\times}10^{10}$ to $3.8{\times}10^{12}$ and $1.7{\times}10^6$ to $2.7{\times}10^{10}$ copies/l, respectively. Activated sludge from livestock, textile, and sewage treating WWTPs contained relatively high amoA gene copies (more than $10^5$ copies/l), whereas activated sludge from food and paper WWTPs revealed a low number of the amoA gene (less than $10^3$ copies/l). The value of the amoA gene copy effectively showed the difference in composition of bacteria in different activated sludge samples and this was better than the measurement with the AOB 16S rRNA or total 16S rRNA gene. These results suggest that the quantification of the amoA gene can help monitor AOB and ammonia oxidation in WWTPs.

This study explored the chronic effects of different doses of the triazole fungicide tebuconazole on the growth, and metabolic and enzymatic functions of the filamentous paddy field cyanobacterium, Westiellopsis prolifica Janet. The growth of the cyanobacterium was determined by an estimation of the change in pigment contents. Chlorophyll-a, carotenoids, and accessory pigments such as phycocyanin, allophycocyanin, and phycoerythrin were shown to decline over a 16-day period by a factor of 92%, 93%, 83%, 95%, and 100%, respectively, with increasing doses of the fungicide. Metabolic and enzymatic activities were also adversely affected. Over the 16 days, a gradual rise in total phenol content was recorded when Westiellopsis prolifica Janet was treated with 60 ppm of the fungicide, despite the reduction in carbohydrates, proteins, and amino acids by 96%, 92%, and 90%, respectively. Moreover, the enzymes nitrate reductase (NR), glutamine synthetase (GS), and succinate dehydrogenase (SDH) also registered reductions of 93%, 90%, and 98%, respectively. This study indicates that tebuconazole, although an important fungicide used extensively in rice fields, exhibits an inhibitory effect on the growth and metabolic activities of Westiellopsis prolifica Janet and hence possibly on other varieties as well.

Differences in DNA banding patterns, obtained by ribosomal intergenic spacer analysis (RISA), and nitrification were followed in a moving-bed biofilm reactor (MBBR) receiving municipal landfill leachate. Complete nitrification (>99%) to nitrate was obtained in the two-stage MBBR system with an ammonium load of 1.09 g N-$NH_4/m^2{\cdot}d$. Increasing the ammonium load to 2.03 g N-$NH_4/m^2{\cdot}d$or more caused a decline in process efficiency to 70-86%. Moreover, at the highest ammonium load (3.76 g N-$NH_4/m^2{\cdot}d$), nitrite was the predominant product of nitrification. Community succession was evident in both compartments in response to changes in ammonium load. Nonmetric multidimensional scaling (NMDS) supported by similarity analysis (ANOSIM) showed that microbial biofilm communities differed between compartments. The microbial biofilm was composed mainly of ammonia-oxidizing bacteria (AOB), with Nitrosomonas europeae and N. eutropha being most abundant. These results suggest that high ammonium concentrations suit particular AOB strains.

In order to establish an axenic (bacteria-free) culture of Microcystis aeruginosa NIER 10039 isolated from a Korean reservoir, the culture was subjected to sequential treatment, including ultrasonication, washing, and addition of antibiotics. Three broad-spectrum antibiotics, namely, kanamycin, ampicillin, and imipenem, were applied separately in that order. Axenicity of the culture was confirmed by cultivation on bacterial media and observation under epifluorescence and scanning electron microscopes. We are the first to establish an axenic culture of a Microcystis strain isolated from Korean reservoirs and can be used in physiological and molecular studies to control toxic Microcystis blooms.

During ethanol fermentation, bacterial strains may encounter various stresses, such as ethanol and acid shock, which adversely affect cell viability and the production of ethanol. Therefore, ethanologenic strains that tolerate abiotic stresses are highly desirable. Bacteria of the genus Deinococcus are extremely resistant to ionizing radiation, ultraviolet light, and desiccation, and therefore constitute an important pool of extreme resistance genes. The irrE gene encodes a general switch responsible for the extreme radioresistance of D. radiodurans. Here, we present evidence that IrrE, acting as a global regulator, confers high stress tolerance to a Zymomonas mobilis strain. Expression of the gene protected Z. mobilis cells against ethanol, acid, osmotic, and thermal shocks. It also markedly improved cell viability, the expression levels and enzyme activities of pyruvate decarboxylase and alcohol dehydrogenase, and the production of ethanol under both ethanol and acid stresses. These data suggest that irrE is a potentially promising gene for improving the abiotic stress tolerance of ethanologenic bacterial strains.