• Microbiology and Biotechnology Letters
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• v.26 no.1
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• pp.55-60
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• 1998

For an ideal extractive bioconversion in aqueous two-phase systems, the product has to be preferentially partitioned into the phase opposite to the one in which the biocatalyst is located. Partitioning behaviors of Lactobacillus helveticus IAM 11090 and lactic acid in aqueous two-phase systems composed a polycation, poly(ethylenimine) (PEI), and an uncharged polymer (hydroxyethyl)cellulose (HEC) were investigated. L. helveticus cells were preferentially partitioned to the HEC-rich top phase while about 85% of lactic acid was partitioned to the PEI-rich bottom phase. These results indicate that extraction of charged, low molecular weight products in an aqueous two-phase systems can be promoted by using an oppositely charged polymer as one of the phase-forming polymer. By the ideal partitioning of the cells and lactic acid, an aqueous PEI/HEC two-phase system can be used as a potential system for the extractive lactic acid fermentation of cheese whey.

• Molecular & Cellular Toxicology
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• v.4 no.4
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• pp.307-310
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• 2008

AAD16034 is an alginate lyase from Pseudoalteromonas sp. IAM14594. A very close homologue with known 3D structure exists (marine bacterium Pseudoalteromonas sp. strain no. 272). A three-dimensional structure of AAD16034 was generated based on this template (PDB code: 1J1T) by comparative modeling. The modeled enzyme exhibited a jelly-roll like structure very similar to its template structure. Both enzymes possess the characteristic alginate sequence YFKhG+Y-Q. Since AAD16034 displays enzymatic activity for poly-M alginate, docking of a tri-mannuronate into the modeled structure was performed. Two separate and adjacent binding sites were found. The ligand was accommodated inside each binding site. By considering both binding sites, a plausible binding pose for the poly-M alginate polymer could be deduced. From the modeled docking pose (i.e., the most important factor that attracts alginate polymer into this lyase) the most likely interaction was electrostatic. In accordance with a previous report, the hydroxyl group of Y345 was positioned close to the ${\alpha}$-hydrogen of ${\beta}$-mannuronate, which was suitable to initiate a ${\beta}$-elimination reaction. K347 was also very near to the carboxylatemoiety of the ligand, which might stabilize the dianion intermediate during the ${\beta}$-elimination reaction. This implies that the characteristic alginate sequence is absolutely crucial for the catalysis. These results may be exploited in the design of novel enzymes with desired properties.

• Analytical Science and Technology
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• v.30 no.1
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• pp.26-31
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• 2017

Background and purpose: This study was carried out to learn more about the potential prophylactic or antibacterial activity of the plant Acalypha indica against selective pathogenic bacteria. Experimental: The test organisms were Sarcina lutea IFO 3232, Bacillus subtilis IFO 3026, Pseudomonas denitrificans, Escherichia coli IFO 3007, Klebsiella pneumoniae ATTC 10031, Xanthomonas campestris IAM 1671, and Proteus vulgaris. Leaf, stem, and bud powder of Acalypha indica were dissolved in various solvents, and the extracts were tested for antimicrobial activity through the disc diffusion method. GC-MS profiling was performed to characterize active chemical compounds in the essential oil of Acalypha indica. Results: The ethanol extract showed the highest activity against all bacteria, while the petroleum ether extract yielded the highest zone of inhibition against Proteus vulgaris ($11.83{\pm}1.75mm$). The minimum inhibitory concentration (MIC) of the ethyl acetate extract against Bacillus subtilis was 16 µg/mL. Phytochemical screening by GC-MS revealed a total of 12 bioactive compounds. Conclusion: Extracts of Acalypha indica may be useful in formulating and synthesizing new antibacterial drugs.

• Korean Journal of Food Science and Technology
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• v.16 no.1
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• pp.41-46
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• 1984

The pectic enzymes produced from Aspergillus niger were separated into three fractions (F-A, F-I and F-II) by means of Sephadex and DEAF-Sephadex column chromatography. Each enzyme fraction was characterized by determining viscosity change and reducing surgar of the pectic acid-enzyme mixture and analyzing thin layer chromatogram of the reaction products. F-I rapidly reduced the viscosity of pectic acid solution and released reducing groups in a random manner so that appeared to be an endo-polygalacturonase (endo-PG). The optimum pH of endo-PG for viscosity reducing activity was 4.2 and that for releasing reducing surgar was 4.7. In the thermal inactivation of endo-PG of $30-45^{\circ}C$, the enthalpy of activation was 217.3 kj/mole and z-value was $7.5^{\circ}C$. F-II and F-A were determined as endo-polymethylgalacturonase and exo-polygalacturonase, respectively.

• Archives of Pharmacal Research
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• v.3 no.1
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• pp.7-12
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• 1980

Ampliciline was synthesized from 6-amino-pencillanic acid (6-APA) and D-.alpha. phenylglycine methyl ester by using amplicilin synthesizing enzyme from Peudomonas melanogenum (IAM 1655). The whole cell enzyme was immobilized by entrapping it in the polyacrylamide gel lattices. The polymer used in the enzyme entrapment was made from 150 mg per ml of acrylamide monomer and 8 mg per ml of N, N'-methylenebisacrylamide. About 200 mg/whole cell enzyme was mixed in the polymer for entrapment. The maximal activity retention after immobilization was 56%. The optimal pH values for the whole cell enzyme and the immobilized whole cell enzyme were 6.0 and 5.9, respectively. The optimal temperature for the enzyme activity were the same for both type of preparations. The enzyme stabilities against pH and heat increased for immobilized whole cell enzyme. Immobilized cell was more stable especially in the acidic condition while both type were found to be very suceptible to thermal inactivation at a temperature above 4.deg.C. The kinetic constants obtained from Lineweaver-Burk plot based on two substate reaction mechanism showed somewhat higher value for immobilized whole cell enzyme as compared to the whole cell enzyme : the Km value for 6-APA were 7.0 mM and 12.5 mM while Km values for phenylglycine methyl ester were 4.5 mM and 8.2 mM, respectively. Using the immobilized whole cell enzyme packed in a column reactor, the productivity of ampiciline was studied by varying the flow rate of substrate solution. At the space velocity, SV, 0.14 hr$^{-1}$ the conversion was 45%. Operational stability found in terms of half life was 30 hr at SV = 0.2 hr.

• Korean Journal of Veterinary Research
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• v.54 no.1
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• pp.39-48
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• 2014

The prevalence of thermophilic Campylobacter (C.) spp. in stray, breeding, and household dogs was 25.2, 12.0, and 8.8%, respectively. C. jejuni and C. upsaliensis were the predominant Campylobacter spp. from household dogs. cdtA, cdtB, and cdtC were detected by PCR in all isolates. Despite the high cytolethal distending toxin (CDT) gene prevalence, only 26 (31%) C. jejuni strains and one (15.3%) C. coli strain showed evidence of CDT production in HEp-2 cell cytotoxicity assays. Virulence-associated genes detected in the C. jejuni and C. coli isolates were cadF, dnaJ, flaA, racR, ciaB, iamA, pldA, virB11, ceuE, and docC. cadF, dnaJ, flaA, and ceuE were found in all C. jejuni and C. coli isolates. When detecting Guillain-Barr$\acute{e}$ syndrome-associated genes (galE, cgtB, and wlaN), galE was identified in all isolates. However, cgtB and wlaN were more prevalent in C. jejuni isolates from humans than those from dogs. Adherence and invasion abilities of the C. jejuni and C. coli strains were tested in INT-407 cells. A considerable correlation (adjusted $R^2$= 0.678) existed between adherence and invasion activities of the Campylobacter spp. isolates.

• KSBB Journal
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• v.6 no.2
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• pp.143-146
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• 1991

The liquifying $\alpha-amylase$ production from B. subtilis, A. oryzae and B. natto using wheat and rice bran as low cost culture medium was investigated. Among 3 strains, B. natto showed heights productivity of $\alpha-amylase$ in the outer wheat bran medium. And the optimum culture condition is pH 6.8 and $37^{\circ}C$ for the production of $\alpha-amylase$. The $\alpha-amylase$ activity of the crude enzyme and the purified enzyme are 256 unit/ml and 10,700 unit/ml, respectivitly. The $\alpha-amylase$ from B. natto cultrtured in outer wheat bran medium was purified into nearly a pure state(98.7%). And the molecular weight of the purified $\alpha-amylase$ was 34,000.

• Microbiology and Biotechnology Letters
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• v.43 no.2
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• pp.134-141
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• 2015

An agarolytic marine bacterium (H9) was isolated from the coastal seawater of the West Sea, South Korea. The isolate, H9, was gram-negative and rod-shaped with a smooth surface and polar flagellum. Cells grew at 20-30℃, between pH 5.0 and 9.0, and in ASW-YP (Artificial Sea Water-Yeast extract, Peptone) media containing 1-5% (w/v) NaCl. The G+C content was 41.56 mol%. The predominant isoprenoid quinone in strain H9 was ubiquinone-8. The major fatty acids (>10%) were C_16:1ω7c (34.3%), C_16:0 (23.72%), and C_18:1ω7c (13.64%). Based on 16S rRNA gene sequencing, and biochemical and chemotaxonomic characterization, the strain was designated as Pseudoalteromonas sp. H9 (=KCTC23887). In liquid culture supplemented with 0.2% agar, the cell density and agarase activity reached a maximum level of OD = 4.32 (48 h) and OD = 3.87 (24 h), respectively. The optimum pH and temperature for the extracellular crude agarases of H9 were 7.0 and 40℃, respectively. Thin-layer chromatography analysis of the agarase hydrolysis products revealed that the crude agarases hydrolyze agarose into neoagarotetraose and neoagarohexaose. Therefore, the new agar-degrading strain, H9, can be applicable for the production of valuable neoagarooligosaccharides and for the complete degradation of agar in bio-industries.

• Applied Biological Chemistry
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• v.36 no.4
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• pp.277-283
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• 1993

In order to brew foxtail millet wine, a folk wine of Cheju, properties of raw materials, optimum brewing conditions were inveatigated. Carbohydrate and crude fat content of glutinuous foxtail millet are 71.27% and 3.47%, respectively. Since the ratio of water to steamed millet and ethanol concentration of wine showed negative correlation, less than 250% water had to added to steamed millet to maintain ethanol concentration in wine above 13%, Sugar consumption and ethanol production increased rapidly for the first 2 days, and main fermentation was done in 4 days. Ethanol concentrations were $13.0{\sim}13.4%$ when foxtail millet was used, and they were $14.0{\sim}14.3%$ for the mixture substrates of 90% millet and 10% rice or barley. Organic acids in millet wine were lactic acid, malic acid and succinic acid. The residual carbohydrates after fermentation were mainly xylose and oligosaccharides. A trace of methanol was detected in millet wine. The content of fusel oil was low, while the concnetration of organic acids was high. Optimum conditions for millet wine-making were as follows. Glutinuous foxtail millet with 10% rice as fermentation source need to be soaked in water and steamed for enough time. Water was added to steamed millet with the ratio of 2 : 1. The resulting mixture was stmnultaneously saccharified and fermented by Aspergillus orzae and Saccharomyces cerevisiae IAM 4274 at $20^{\circ}C$ for a week. Millet wine was prepared after filtering fermented broth while pressing for a week.

• Korean Journal of Microbiology
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• v.17 no.4
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• pp.165-178
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• 1979

Phages of Lactobaciilus casei (PLC) isolated from plant drainage were classified and characterized. The results are as follows : 1. On the basis of host range pattern, phages could be divided into 2 groups (PLC-B and PLC-C). PLC-B group phages could be further divided into 5 sub-groups $(B_1, \;B_2, \;B_3, \;B_4, \;and\;B_5)$. Although PLC-C group phages had the same host range, they could be also divided into 2 sub-groups $(C_1\;and\;C_2)$ by morphlogical type. 2. It was $B_3$ group phages that represented a major proportion (44.4%) of phages tested. However, $B_1$ group phages were shown to have the widest host range. 3. Electron micrographs revealed that the phages fell into three different morphological types. $(B_1, \;B_2, \;and\;B_3)$ group phages hd a hexagonal head (52nm in diameter) and a sheathless noncontractile (245 nm in length). $B_4\;and\;C_2$ group phages had a hexagonal head (56 nm) and a short flexible tail (169nm) having no sheath. $B_5\;and\;C_1$ group phages were shown to have a hexagonal head (81 nm) and a contractile tail (140 nm) having a sheath, a base plate and tail fibers. 4. The inactivation of the phages by antisera indicated that serological relationships correlated completely with morphological types. 5. $B_1, \;C_1\;and\;C_2$ group phages produced a large (1, 2 mm in diameter) plaque with a clear ring. The morphology of plaques of $B_3\;and\;B_5$ group phages was the same as those produced by the above, but the average plaque sizes for $B_3\;and\;B_5$ were 0.8 mm abd 0.5 mm, respectively. $B_2\;and\;B_4$ group phages produced a small (0.5 mm) turbid plaque with an irregular edge. 6. The latent period and the average burst size of $B_1\;and\;B_3$ group phages were 90 min and 100, respectively. These phages reuqired calcium ions for their miltiplication. 7. $B_3$ group phages could not be absrobed to R-variant $KC_1$. 8. The order of resistance of phages to heat was $B_2\;>\;B_1, B_4\;and\;B_5\;>\;B_3\;and\;C_2, \;B_5$ group phages were more stable than $B_3$ in various pH values. $C_2$ group phages were more sensitive to UV irradiation than $B_1\;and\;B_3$ group phages. 9. Strains YIT9018 and IAM 1043 were induced by mitomycin C treatment. Phage particles detected in the lysates had a hexagonal head (38 and 49 nm, respectively), but no tail. Any sensitive indicator strain could not be isolated in spite of repaeated trials.