Seven isolates showing strong proteolytic activity, KYC 1028, 1100, 1134, 1139, 1151, 1159, and 1182, were collected. Out of them, the broth of KYC 1134 and KYC 1139 showed the high proteolytic activity measured by azocazein. To determine 16S rDNA sequences for identification, 16S rDNA of seven isolates were amplified and compared with the 16S rDNA sequences of other myxobacteria at NCBI. It is evident from the phylo-genetic tree that the isolates belong to the genus Myxococcus. Sharing high percentage similarity values with myxobacteria, the 16S rDNA sequences were involved in two species, Myxococcus macrospores and M. Fulvus. Biochemical characteristics of KYC 1134 broth, which showed the highest proteolytic activity, showed increased activity 8 times to seven days after culture, and protein production were increased gradually and stopped at five days. The broth had optimal temperature at for proteolytic activity, and stability of pH was ranged from pH 5 to 10, at and 60, respectively. To classify proteases being in the broth, ten inhibitors were determined and only bestatin showed 27% inhibition effect. The inhibition result demonstrates that the broth contains kinds of amino peptidases and other exopeptidases.

Bacteria were isolated from roots of plants belonging to family Solanaceae and Gramineae, inhabited in Dokdo island. Fifty six endospore-forming bacteria grown on tryptic soy broth (TSB) agar medium and 23 nitrogen-fixing bacteria (NFB) grown on nitrogen free agar medium were isolated, respectively. The isolates were partially identified by analyzing the 16S rDNA and categorized into phylogenetic groups. The 16S rDNA sequences of each identified isolates were compared with sequences of each type strains to analyze phylogenetic relationship by phylogenetic tree. As a result, endospore-forming bacteria and nitrogen-fixing bacteria were classified into 4 and 6 lineage groups, respectively. Among these isolated, 18 were presumed to be novel species candidates based on the similarity (lower than 98%) analysis of the l6S rDNA sequences.

An indole oxygenase originated from Rhodococcus sp. RHA1 was cloned into the expression vector, pTrc99A, in Escherichia coli, and designated pTCAN1. The pTCAN2 was constructed from pTCAN1 by the deletion of for the constitutive expression of indole oxygenase without adding IPTG in the medium. The complete open reading frame of indole oxygenase was 1,224 bp long, which encodes a protein of 407amino acids. Crude extracts of E. coli /pTCAN1 and pTCAN2, respectively, were prepared and subjected to SDS-PAGE analysis. A band corresponding to molecular mass of about 43 kDa was appeared and this result correlated with the predicted molecular mass of cloned indole oxygenase. The E. coli harboring pTCAN1 and pTCAN2, respectively, showed blue color colony in LB plate. The pigment showing blue color was prepared from E. coli /pTCAN2, and identified as indigo by experiments using spectrophotometer, HPLC, and TLC. The indigo-forming activity of indole oxygenases from the whole cell of E. coli cultured at LB medium added 1mM of IPTG and that of E. coli/pTCAN2 showed about 1.75nmol/min/mg DCW (dry cell weight) and 3.85 nmol/min/mg DCW, respectively. Also, the E. coli /pTCAN2 produced about of indigo after 48 hours incubation in TB medium supplemented with 2.5 mM of tryptophan.

A strain producing -galactosidase and -glucosidase was isolated from Kimchi. The isolated strain was identified as Weissella cibaria by 16S rDNA analysis and designated as Weissella cibaria K-M1-4. The enzyme activity of -galactosidase and -glucosidase reached the maximum in the soy medium at for 24 hr. The enzymes were purified by ethanol fractionation, DEAE sepharose fast flow, and sephacryl S-100HR column chromatography. -Galactosidase specific activity was shown by 576 Units/mg protein and the yield was 3.5% of the total activity of crude extracts. -glucosidase specific activity was shown by 480 Units/mg protein and the yield was 2.9% of the total activity of crude extracts. The optimum temperature for -galactosidase was and 43% of its original activity remained when it was treated at for 30 min. For -galactosidase shows the optimum pH of 8.0 and is fairly stable between pH5.0 and pH9.0. The enzyme activity was increased in the presence of and . The value of Km and Vmax for the enzyme were 0.98 mM and mole/min, respectively. The -glucosidase has the optimum temperature of and 46% of its original activity remained when it was treated at for 30min. Its optimum pH of 7.0 and is fairly stable between pH5.0 and pH9.0. The enzyme activity was increased in the presence of and . The value of Km and Vmax for the enzyme were 1.24 mM and mole/min, respectively.

The goal of this study was to develop new functional pear wine using six Asian pears (Pyrus pyrifolia, Nakai), namely Wonhwang, Niitaka, Whangkeumbae, Whasan, Gamcheonbae and Chuwhangbae. To select optimal yeast and pear, we investigated the physicochemical properties of the pear wines from fermentation of musts of six pear cultivars at for 7 days by several yeasts. of ethanol from musts of 'Wonhwang', 'Whangkeumbae' and 'Whasan' were produced by Saccharomyces cerevisiae K-7 and 12.8% of ethanol was also produced from 'Niitaka' by commercial Saccharomyces cerevisiae C-2. 9.9% and 11.4% of ethanol were produced from musts of 'Gamcheonbae' and 'Chuwhangbae' by Saccharomyces cerevisiae KCTC 7904, respectively. Among several pear wines, Niitaka pear wine showed the best acceptability in the sensory evaluation, and Niitaka pear wine and Whangkeumbae pear wine showed 31.1% and 27.8% of antihypertensive angiotensin I-converting enzyme(ACE) inhibitory activity, respectively. However, the other functionalities were not detected or very low. Furthermore, Niitaka-strawberry mixed fermentation wine was showed the excellent acceptability and high antihypertensive ACE inhibitory activity of 64.9%.

A bacterial strain, JE-34, producing a high concentration of red pigment was isolated from a sediment in East China Sea. It was identified as Zooshikella sp. JE-34 based on the 16S rRNA gene sequence analysis. The red pigment was purified by solvent extraction and HPLC was identified as prodigiosin-like compound. Nutritional and cultural conditions were optimized for the production of prodigiosin-like pigment in the flask level. Optimal culture conditions were at initial medium pH , and 4 days incubation. For carbon and, nitrogen sources were soluble starch and malt extract.

Rice straw was pretreated using dilute sulfuric acid at reaction conditions covering two levels of reaction temperature (140, ) and five levels of acid concentrations (). The production and decomposition rates of major components of rice straw indicating glucose, xylose, galactose and arabinose were investigated. The production rate of arabinose and the decomposition rate of xylose were greatest among them. The maximum attainable hemicellulose (xylose+galactose+arabinose) yield was about 80%. High acid concentration appears to favor the maximum yield but high temperature does not. The optimum condition was found to be , 2.5% and 20 minutes. The maximum glucose yields were almost same, around , regardless of reaction conditions.

This paper aimed to develop a solvent extraction and purification process to recover high-purified -carotene from recombinant Escherichia coli. Cells harvested from the culture broth were treated through numerous steps: dehydration, solvent extraction, crystal formation and separation. To optimize the extracting condition, experiments were carried out to investigate the effect of cell disruption, temperature, organic solvents, solvent-biomass ratio on the yield of -carotene extracted from cells. The result indicated that no significant differences of extraction yield were observed from cells with or without step of cell disruption. Among different extracting solvents, the highest extraction yield of -carotene, 30.3 mg--carotene/g-dry cells, was obtained with isobutyl acetate at solvent-biomass ratio 25 mL/g-dry cells at . Notably, in case of acetone, the extraction yield was quite low when using acetone itself, but increased almost up to the highest value when combining this solvent and olive oil. The purity of -carotene crystals obtained from crystallization and separation was 89%. The purity degree was further improved up to 98.5% by treating crude crystals with additional ethanol washing.

Pediococci selective medium (PSM) supplemented with ampicillin (A) reported as valid for the detection and enumeration of pediococci included in foods and animal feed was evaluated for the selective detection of the genus Pediococcus in kimchi. PSM is based on the complex basal medium MRS supplemented with cysteine hydrochloride, vancomycin, novobiocin, and nystatin. In the medium evaluation with known species, the growth inhibition of leuconostocs, Pediococcus pentosaceus, Lactobacillus casei, Lactobacillus curvatus, Oenococcus oeni, and Streptococcus thermophilus was not confirmed. In the application of kimchi samples on the selective medium, leuconostocs, P. pentosaceus, Weissella koreensis, Lb. curvatus, Lactobacillus brevis, and Lactobacillus sakei were detected. PSM+A was proved to be not applicable for the detection of pediococci in kimchi.

The usage of ethanol (EtOH)-blended gasoline (gasohol), has been increasing in recent years. EtOH has influence on the distribution and biodegradation of aromatic compounds such as BTEX (benzene (B), toluene (T), ethylbenzene (B), and xylene (X)) that are gasoline compositions. In this study, the effect of EtOH on the aerobic biodegradation of B, T and E was investigated using a BTEX and EtOH-degrading bacterium, Rhodococcus sp. EH831. The degradation rates of B in the conditions of 1:1, 1:4, and 1:0.25 mixtures with EtOH (B:EtOH, mol:mol) were ranged from to cell wight . The degradation rate of T was the fastest in the 1:0.25 mixture (), and it was the lowest in the 1:4 mixture (). The degradation rates of E were increased with increasing the addition amount of EtOH: The degradation rate of E was the highest in the 1:4 mixture (), and the rates were , , and in the 1:1, 1:0.25, 1.0 mixtures, respectively. In conclusion, the biodegradation of B, T, E by Rhodococcus sp. EH831 was not significantly inhibited by the co-existence of EtOH.

The inhibitors against Vibrio harveyi quorum sensing (QS) signaling were developed by modifying the molecular structure of the major signal, N-3-hydroxybutanoyl-L-homoserine lactone (3-OH--HSL). A series of structural derivatives, N-(3-hydroxysulfonyl)-L-homoserine lactones (HSHLs) were synthesized by the solid-phase organic synthesis method. The in vivo QS inhibition by these compounds was measured by a bioassay system using the V. harveyi bioluminescence, and all showed significant inhibitory effects. To analyze the interaction between these compounds and LuxN, a 3-OH--HSL receptor protein of V. harveyi, we tentatively determined the putative signal binding domain of LuxN based on the sequence homology with other acyl-HSL binding proteins, and predicted the partial 3-D structure of the putative signal binding domain of LuxN by using ORCHESTRA program, and further estimated the binding poses and energies (docking scores) of 3-OH--HSL and HSHLs within the domain. In comparison of the result from this modeling study with that of in vivo bioassay, we suggest that the in silica interpretation of the interaction between ligands and their receptor proteins can be a valuable way to develop better competitive inhibitors, especially in the case that the structural information of the protein is limited.

A study on livestock environment improving agents was conducted; top two brands (A and B) in the market, bottom two brands (E and F) based on market shares and two newly developed agents (C and D) were measured for viable count and tested for resistance towards antibiotics prohibited against livestock feeds. Test results revealed that the measured viable count of agents A and B matched those on the labels were identical; however agent E lacked information on viable counts nor the intended usage, while the measured viable count of agent F was less than the label-stated count. No correlation was found between the antibiotic-resistance test and market share, and most of the agents excluding B were found to display resistance case of Lincosimides such as Lincomycine and Clindmycin, resistant bacteria were found, with the except of agent B. Amoxicillin, Ampicillin and Penillin (type-Penecillins) and Erythromycin (type-Macrolide) were shown to contain resistant bacteria, with the except of agents Band E; the same for Norploxacin (type-Quinoline) and Neomycin antibiotics. Aminoglycosides such as Gentamycin and Streptomycin contained resistant bacteria, excluding agent B. Oxytetracyclin (type-Tetracycline), which is banned for use as resistant bacteria showed the highest sensitivity among the 12 antibiotics, revealed positive results in the test for resistant bacteria; again excluding of agents Band E. These results reveal that many agents contained resistant bacteria despite the fact that they were prohibited; this calls for a more accurate display of the facts and specifications, systematic distributions and strict verification processes of environment improving agents.

The annual productions of yam and its aerial bulbils are estimated to 5,000 and 2,500 ton, respectively. But the majority of bulbils had been discarded without specific use. In this study, methanol extract and its organic solvent fractions were prepared from bulbils of Dioscorea batatas Decne, and their antimicrobial, antithrombin, and antioxidant activities were evaluated, respectively. The methanol extract contained 58.98% of water-soluble materials as like yam's extract. But the bulbils's extract contained 12-folds of total polyphenol and 3.4-folds of total flavonoids compared than yam's extract, respectively. For antimicrobial activity the hexane and ethyl acetate fraction showed strong antibacterial activity at concentration against gram-negative and gram-positive bacteria. Antifungal activity was not observed in any fractions. Strong antithrombin activity was found in the hexane fractions. At 4.8 mg/mL concentration thrombin time (TT) was over 300 sec, which is 4-folds extended than the TT of yam. In a while, the ethyl acetate fraction showed strong DPPH scavenging activity ( of ), SOD-like activity and reducing power, which are comparable to vitamin C or BHT. Our results suggest that the bulbils of yam as yam tuber have useful bio-activities, such as antibacterial, antithrombosis, and antioxidant activity.

Hyungbangjihwang-Tang (HT), an Oriental herbal formula, has been known to play a role which helps to recover vigor of human in the Orient. In this study, antifungal substance (HTI) was purified from the ethyl-acetate extracts of HT by using column chromatography and HPLC, and the antifungal effects of HTI and its mode of action were investigated. By using a broth micro-dilution assay, the activity of HTI was evaluated against fungi. HTI showed antifungal activities without hemolytic effect against human erythrocytes. To confirm antifungal activity of HTI, we examined the accumulation of intracellular trehalose as stress response on toxic agents and effect on dimorphic transition in Candida albicans. The results demonstrated that HTI induced the accumulation of intracellular trehalose and exerted its antifungal effect by disrupting the mycelial forms. To understand its antifungal mode of action, cell cycle analysis was performed with C. albicans, and the results showed HTI arrested the cell cycle at the S phase in yeast. The present study indicates that HTI has considerable antifungal activity, deserving further investigation for clinical applications.

To investigate anti-phytopathogenic fungal activity of Coptis chinensis, the methanol extract and its organic solvent fractions were prepared. Using the extract and the fractions, in-vitro spore-germination inhibition and mycelial-growth inhibition activities were evaluated against Colletotrichum gloeosporioides, Phytohpthora capsici, Pyricularia grisea, Rhizoctonia solani, Botryosphaeri dothidea, Glomerella cingulata, respectively. Treatment of the methanol extract (500 mg/mL) into the spore of phytopathogenic fungi completely inhibited germinations for 5 days, except B. dothidea, and showed strong antifungal activities against P. grisea and B. cinerea, and antioomycetes activity against P. capsici. The minimal growth inhibition concentrations of the methanol extract against P. grisea, B. cinerea and P. capsici were 300, 300, and 500 mg/mL, respectively. For practical application of C. chinensis in red-pepper field, the hot-water extract (1,000 mg/mL) was prepared in commercial facility, after evaluation of heat stability and solvent-extraction yields of antifungal substances. The 3-times leaf-spray of the extract from June to August, 2008 did not show any deleterious effect to red-pepper. In fact, the leaf-spray promoted plant growth including leaf, root and fruit. The average weight and rind of each fruit were increased to 119% and 117% comparison to those of without treatments. Our results suggest that C. chinensis is a useful source for control of red-pepper diseases and plant growth.

The nucleocapsid (N) protein of porcine reproductive and respiratory syndrome virus (PRRSV) is a basic multifunctional protein which has been reported to be a serine phosphoprotein with yet-identified functions. As a first step towards understanding the general role of N protein phosphorylation during virus replication, the non-phosphorylated mutant N gene was constructed by mutating all serine residues to alanine. This recombinant N protein was identified to be unphosphorylated, confirming that serine residues truly function as core amino acids responsible for N protein phosphorylation. The PRRSV N protein has been shown to possess the biological features of nuclear localization and N-N homodimerization which individually play critical roles in virus infection. In the present study, therefore, it was attempted to investigate whether these two properties of the N protein are modulated by its phosphorylation status. However, experimental results showed that the non-phosphorylated N protein was still present in the nucleus and nucleolus, and was able to associate with itself by non-covalent interactions. Taken together, the data suggest phosphorylation-independent regulation of N protein nuclear transport or oligomerization, thereby implying the potential involvement of phosphorylation in regulating the activities of the N protein at other levels including RNA-binding capacity.