• Journal of Life Science
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• v.27 no.2
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• pp.202-210
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• 2017

As an effort to find a potential biopreservative, we isolated bacterial strains producing bacteriocin from fermented foods. A strain was finally selected and characteristics of the bacteriocin were investigated. The selected strain was identified as Bacillus subtilis E9-1 based on the 16S rRNA gene analysis. The culture supernatant of B. subtilis E9-1 showed antimicrobial activity against Gram-positive bacteria. Subtilisin A, ${\alpha}$-chymotrypsin, trypsin and proteinase K inactivated the antimicrobial activity, which means its proteinaceous nature, a bacteriocin. The bacteriocin activity was fully retained at the pH range from 2.0 to 8.0 and stable at up to $100^{\circ}C$ for 60 min. Solvents such as ethanol, isopropanol and methanol had no effect on the antimicrobial activity at the concentration of 100% but acetone and acetonitrile reduced the activity at up to 100% concentration. Cell growth of four indicator strains was dramatically decreased in dose-dependent manner. Listeria monocytogenes was the most sensitive, but Enterococcus faecium was the most resistant. Bacillus cereus and Staphylococcus aureus showed the medium sensitivity. The bacteriocin showed its antimicrobial activity against B. cereus and L. monocytogenes via bactericidal action. The number of viable cells of L. monocytogenes started to reduce after addition of bacteriocin to the minced beef. The bacteriocin was purified through acetone concentration, gel filtration chromatography and RP-HPLC. The whole purification step led to a 6.82 fold increase in the specific activity and 6% yield of bacteriocin activity. The molecular weight of the purified bacteriocin was determined to be 3.3 kDa by MALDI-TOF/TOF mass spectrometry.

• Korean Journal of Food Science and Technology
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• v.9 no.4
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• pp.251-254
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• 1977

Pectin, hesperidin and naringin were extracted from hot air-dried peel ($60^{\circ}C$, 1 hr and air velocity 160 fpm) of citrus produced in Korea in order to see the amount of each component contained in the peel. Pectin was extracted by three different methods and the quality and contents of the pectins were determined respectively. 1. The pectin yield by the total pectic substance method was the highest (26.0% for unshiu (U) and 28.5% for natsudaidai (N) expectedly and the soluble pectic substance method the least (13.5%(U) and 15.6% (N)) The yield by method III (extraction by water at pH 1.5 followed by isopropanol precipitation) was intermediate (18.1% (U) and 20.8%(N)). Anhydrouronic acid (AUA) content was the highest (92.0% (U) and 90.3%(N)) in those by method III. The AUA contents of the other pectins were 80.0% for soluble pectin (for both U and N), 71.6% for the commercial pectin (Sunkist Groups Inc., U.S.A.), 58.0%(N) and 63.4%(U) for total pectic substance. 2. The methoxyl content of total pectic substance was the lowest (4.81%(U) and 4.88%(N)). However, there was no significant difference in methoxyl content among the rest which were found to have low levels(5.27-7.20%). 3. The pectin by method III gave the highest jelly strength. The commercial pectin, soluble pectice substance and total pectic substance were next in order. 4. The hesperidin content of unshiu was 5.07% (dry basis) and the naringin content of natsudaidai 3.03% (dry basis).

• Journal of Food Hygiene and Safety
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• v.20 no.4
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• pp.267-271
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• 2005

Lecithin is a naturally occurring group of phospholipids found in nearly every living cell and has been widely used as the ingredient of functional foods. Lecithin has high content of phosphatidylcholine(PC), pharmaceutical material which promotes metabolism through the cell membrane. This study was carried out to improve the present inconvenient analytical method of PC in law for health & functional foods. The commodities used in this experiment, were two kinds of egg yolk and eight kinds of soybean lecithin functional foods. PC was separated with isocratic elution with hexane : isopropanol : D.W (30:60:8) through silica column (2.1$\times$150 mm) by HPLC with Evaporative Light-Scattering Detector (ELSD). The flow rate of the eluent was 0.5 ml/mim and infect volume was 10ul. The neubilizer temperature of detector was $60^{\circ}C$, drift tube temperature of that was $75^{\circ}C$ and gas flow was 30 psi. Quantification was carried out by external standardization. Limit of quantification was 0.15ppm. Lecithin contents of egg yolk and soybean Products were > $66\%$ and > $81\%$), respectively. Phosphatidylcholine contents of egg yolk and soybean products were > $74\%$ and > $18\%$, respectively.

• Journal of Microbiology and Biotechnology
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• v.25 no.11
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• pp.1880-1893
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• 2015

In this study, Pseudomonas R0-14, which was isolated from Arctic soil samples, showed a clear halo when grown on M9 medium agarose plates containing olive oil-rhodamine B as substrate, suggesting that it expressed putative lipase(s). A putative lipase gene, lipR, was cloned from R0-14 by genome walking and Touchdown PCR. lipR encodes a 562-amino-acid polypeptide showing a typical α/β hydrolase structure with a catalytic triad consisting of Ser₁₅₃-Asp₂₀₂-His₂₆₀ and one α-helical lid (residues 103-113). A phylogenetic analysis revealed that LipR belongs to the lipase subfamily I.3. LipR was successfully expressed in Escherichia coli, purified, and biochemically characterized. Recombinant LipR exhibited its maximum activity towards p-nitrophenyl butyrate at pH 8.5 and 60℃ with a K_m of 0.37 mM and a k_cat of 6.42 s^-1. It retained over 90% of its original activity after incubation at 50℃ for 12 h. In addition, LipR was activated by Ca²⁺, Mg²⁺, Ba²⁺, and Sr²⁺, while strongly inhibited by Cu²⁺, Zn²⁺, Mn²⁺, and ethylenediaminetetraacetic acid. Moreover, it showed a certain tolerance to organic solvents, including acetonitrile, isopropanol, acetone, methanol, and tert-butanol. When algal oil was hydrolyzed by LipR for 24 h, there was an enrichment of n-3 long-chain polyunsaturated fatty acids, including eicosapentaenoic acid (1.22%, 1.65-fold), docosapentaenoic acid (21.24%, 2.04-fold), and docosahexaenoic acid (36.98%, 1.33-fold), and even a certain amount of diacylglycerols was also produced. As a result, LipR has great prospect in industrial applications, especially in food and/or cosmetics applications.

• Journal of Microbiology and Biotechnology
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• v.24 no.6
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• pp.748-755
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• 2014

In the genome annotation of Escherichia coli MG1655, the orf382 (1,149 bp) is designated as a gene encoding an alcohol dehydrogenase that may be Fe-dependent. In this study, the gene was amplified from the genome by PCR and overexpressed in Escherichia coli BL21(DE3). The recombinant $6{\times}$His-tag protein was then purified and characterized. In an enzymatic assay using different hydroxyl-containing substrates (n-butanol, $\small{L}$-threonine, ethanol, isopropanol, glucose, glycerol, $\small{L}$-serine, lactic acid, citric acid, methanol, or $\small{D}$-threonine), the enzyme showed the highest activity on $\small{L}$-threonine. Characterization of the mutant constructed using gene knockout of the orf382 also implied the function of the enzyme in the metabolism of $\small{L}$-threonine into glycine. Considering the presence of tested substrates in living E. coli cel ls and previous literature, we believed that the suitable nomenclature for the enzyme should be an $\small{L}$-threonine dehydrogenase (LTDH). When using $\small{L}$-threonine as the substrate, the enzyme exhibited the best catalytic performance at $39^{\circ}C$ and pH 9.8 with $NAD^+$ as the cofactor. The determination of the Km values towards $\small{L}$-threonine (Km = $11.29{\mu}M$), ethanol ($222.5{\mu}M$), and n-butanol ($8.02{\mu}M$) also confirmed the enzyme as an LTDH. Furthermore, the LTDH was shown to be an ion-containing protein based on inductively coupled plasma-atomic emission spectrometry with an isoelectronic point of pH 5.4. Moreover, a circular dichroism analysis revealed that the metal ion was structurally and enzymatically essential, as its deprivation remarkably changed the ${\alpha}$-helix percentage (from 12.6% to 6.3%).

• Journal of Pharmaceutical Investigation
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• v.35 no.4
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• pp.265-271
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• 2005

A rapid, selective and sensitive reversed-phase HPLC method for the determination of etodolac in human serum was developed, validated, and applied to the pharmacokinetic study of etodolac. Etodolac and internal standard, ibuprofen were extracted from human serum by liquid-liquid extraction with hexane/isopropanol (95:5, v/v) and analyzed on a Luna C18(2) column with the mobile phase of 1% aqueous acetic acid-acetonitrile (4:6, v/v). Detection wavelength of 227 nm and flow rate of 1.0 mL/min were fixed for the study. The assay robustness for the changes of mobile phase pH, organic solvent content, and flow rate was confirmed by $3^3$ factorial design using a fixed etodolac concentration $(1\;{\mu}g/mL)$ with respect to its peak area and retention time. And also, the ruggedness of this method was investigated at three different laboratories using same quality control (QC) samples. This method showed linear response over the concentration range of $0.05-40\;{\mu}g/mL$ with correlation coefficients greater than 0.999. The lower limit of quantification using 0.5 mL of serum was 0.05 ${\mu}g/mL$, which was sensitive enough for pharmacokinetic studies. The overall accuracy of the quality control samples ranged from 92.00 to 110.00% for etodolac with overall precision (% C.V.) being 1.08-10.11%. The percent recovery for human serum was in the range of 76.73-115.30%. Stability studies showed that etodolac was stable during storage, or during the assay procedure in human serum. The peak area and retention time of etodolac were not significantly affected by the changes of mobile phase pH, organic solvent content, and flow rate under the conditions studied. This method showed good ruggedness (within 15% C.V.) and was successfully used for the analysis of etodolac in human serum samples for the pharmacokinetic studies of orally administered Lodin XL tablet (400 mg as etodolac) at three different laboratories, demonstrating the suitability of the method.

• Journal of the Korean Ceramic Society
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• v.42 no.7 s.278
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• pp.475-482
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• 2005

Zn$_{l-x}$Co$_{x}$O (x = 0.05 - 0.20) films were grown on Coming 7059 glass by sol-gel process. A homogeneous and stable Zn$_{l-x}$Co$_{x}$O sol was prepared by dissolving zinc acetate dihydrate (Zn(CH$_{3}$COO)$_{2}$$\cdot$2H$_{2}$O), cobalt acetate tetrahydrate ((CH$_{3}$)$_{2}$$\cdot$CHOH) and aluminium chloride hexahydrate (AlCl$_{3}$ $\cdot$ 6H$_{2}$O) as solute in solution of isopropanol ((CH$_{3}$)$_{2}$$\cdot$CHOH) and monoethanolamine (MEA:H$_{2}$NCH$_{2}$CH$_{2}$OH). The films grown by spin coating method were postheated in air at 650°C for 1 h and annealed in the condition of vacuum (5 $\times$ 10$^{-6}$ Torr) at 300$^{\circ}C$ for 30 min and investigated the nature of c-axis preferred orientation and physical properties with different Co concentrations. Znl_xCOxO thin films with different Co concentrations were well oriented along the c-axis, but especially a highly c-axis oriented Zn$_{l-x}$Co$_{x}$O thin film was grown at 10 at$\%$ Co concentration. The transmittance spectra showed that Zn$_{l-x}$Co$_{x}$O thin films occur typical d-d transitions and sp-d exchange interaction became activated with increasing Co concentration. The electrical resistivity of the films at 10 at$\%$ Co had the lowest value due to the highest c-axis orientation. X-ray photoelectron spectroscopy and alternating gradient magnetometer analyses indicated that no Co metal cluster was formed, and the ferromagnetic properties appeared, respectively. The characteristics of the electrical resistivity and room temperature ferromagnetism of Zn$_{l-x}$Co$_{x}$O thin films suggested the possibility for the application to dilute magnetic semiconductors.

• Journal of the Microelectronics and Packaging Society
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• v.10 no.2
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• pp.1-5
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• 2003

In this project, the efficiency of localized heating for micro systems packaging is developed by using a forced potential scheme microheater. Less than 0.2 Mpa contact pressure was used for bonding with a 200 mA current input for $50{\mu}m$ width, $2{\mu}m$ height and $8mm{\times}8mm$, $5mm{\times}5mm$, $3mm{\times}3mm$ sized phosphorus-doped poly-silicon microheater. The temperature can be raised at the bonding region to $800^{\circ}C$, and it was enough to achieve a strong and reliable bonding in 3 minutes. The IR camera test results show improved uniformity in heat distribution compared with conventional microheaters. For performing the gross leak check, IPA (Isopropanol Alcohol) was used. Since IPA has better wetability than water, it can easily penetrate small openings, and is more suitable for conducting a gross leak check. The pass ratio of bonded dies was 67%, for conventional localized heating, and 85% for our newly developed FP scheme. The bonding strength was more than 25Mpa for FP scheme packaging, which shows that FP scheme can be a good candidate for micro-scale hermetic packaging.

• Journal of Pharmaceutical Investigation
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• v.36 no.1
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• pp.23-29
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• 2006

A rapid, selective and sensitive reversed-phase HPLC method for the determination of promethazine in human serum was developed, validated, and applied to the pharmacokinetic study of promethazine. Promethazine and internal standard, chlorpromazine, were extracted from human serum by liquid-liquid extraction with n-hexane containing 0.8% isopropanol and analyzed on a Capcell Pak CN column with the mobile phase of acetonitrile-0.2 M potassium dihydrogen phosphate (42:58, v/v, adjusted to pH 6.0 with 1 M NaOH). Detection wavelength of 251 nm and flow rate of 0.9 mL/min were fixed for the study. The assay robustness for the changes of mobile phase pH, organic solvent content, and flow rate was confirmed by $3^{3}$ factorial design using a fixed promethazine concentration (10 ng/mL) with respect to its peak area and retention time. In addition, the ruggedness of this method was investigated at three different laboratories using same quality control (QC) samples. This method showed linear response over the concentration range of 1-40 ng/mL with correlation coefficients greater than 0.999. The lower limit of quantification using 1 mL of serum was 1 ng/mL, which was sensitive enough for pharmacokinetic studies. The overall accuracy of the quality control samples ranged from 96.15 to 105.40% for promethazine with overall precision (% C.V.) being 6.70-11.22%. The relative mean recovery of promethazine for human serum was 63.54%. Stability (freeze-thaw and short-term) studies showed that promethazine was stable during storage, or during the assay procedure in human serum. However, the storage at $-80^{\circ}C$ for 4 weeks showed that promethazine was not stable. Extracted serum sample and stock solution were not allowed to stand at ambient temperature for 12 hr prior to injection. The peak area and retention time of promethazine were not significantly affected by the changes of mobile phase pH, organic solvent content, and flow rate under the conditions studied. This method showed good ruggedness (within 15% C.V.) and was successfully used for the analysis of promethazine in human serum samples for the pharmacokinetic studies of orally administered Himazin tablet (25 mg as promethazine hydrochloride) at three different laboratories, demonstrating the suitability of the method.

• Journal of Life Science
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• v.15 no.3 s.70
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• pp.337-343
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• 2005

Tapioca alcoholic distillery waste was utilized as dual purposes to produce citric acid and to reduce the amount of waste to be treated. Primarily an attempt was made to optimize the process conditions by Aspergillus niger in shake bath. The effects of pH, temperature, nitrogen and phosphorus sources on citric acid production were investigated. Maximum concentration of citric acid was made at temperature of $30^{\circ}C$ and pH of 4.3, while maximum cell dry weight was obtained at $35^{\circ}C$. The addition of methanol or ethanol to culture medium promoted citric acid production remarkably, but the addition of $NH_4NO_3,\;KH_2PO_4$ and Manganese as mineral source decreased the acid production.