• Journal of Microbiology and Biotechnology
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• v.33 no.5
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• pp.656-661
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• 2023

The aims of this study were to optimize the preparation of low-molecular-weight collagen using a proteolytic enzyme (alcalase) derived from the feet of Korean native chickens, and to characterize the process of collagen hydrolysis. Foreign bodies from chicken feet were removed using ultrasonication at 28 kHz with 1.36 kW for more than 25 min. The hydrolytic pattern and molecular weight distribution of enzyme-treated collagen from chicken feet were analyzed using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and high-performance liquid chromatography, respectively. Ideally, chicken feet should be treated at 100℃ for 8 h to obtain a high collagen content using hot water extraction. The collagen content of the chicken foot extract was 13.9 g/100 g, and the proportion of low-molecular-weight collagen increased with increasing proteolytic enzyme concentration and reaction time. When treated with 1% alcalase, the average molecular weight of collagen decreased rapidly to 4,929 Da within 5 h and thereafter decreased at a slower rate, reaching 4,916 Da after 7 h. Size exclusion chromatography revealed that low-molecular-weight collagen peptides of approximately 1,000-5,000 Da were obtained after hydrolysis with 1% alcalase for 1 h.

• FOCUS: LIFE SCIENCE
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• no.1
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• pp.1.1-1.4
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• 2024

This article provides comprehensive guidance on the maintenance, cleaning, regeneration, and storage of silica-based HPLC (High-Performance Liquid Chromatography) columns. The general considerations emphasize the importance of using in-line filters and guard cartridges to protect columns from blockage and irreversible sample adsorption. While these measures help, contamination by strongly adsorbed sample components can still occur over time, leading to an increase in back pressure, loss of efficiency, and other issues. To maximize column lifetime, especially with UHPLC (Ultra-High Performance Liquid Chromatography) columns, it is advisable to use ultra-pure solvents, freshly prepared aqueous mobile phases, and to filter all samples, standards, and mobile phases. Additionally, an in-line filter system and sample clean-up on dirty samples are recommended. However, in cases of irreversible compound adsorption or column voiding, regeneration may not be possible. The document also provides specific recommendations for column cleaning procedures, including the flushing procedures for various types of columns such as reversed phase, unbonded silica, bonded normal phase, anion exchange, cation exchange, and size exclusion columns for proteins. The flushing procedures involve using specific solvents in a series to clean and regenerate the columns. It is emphasized that the flow rate during flushing should not exceed the specified limit for the particular column, and the last solvent used should be compatible with the mobile phase. Furthermore, the article outlines the storage conditions for silica based HPLC columns, highlighting the impact of storage conditions on the column's lifetime. It is recommended to flush all buffers, salts, and ion-pairing reagents from the column before storage. The storage solvent should ideally match the one used in the initial column test chromatogram provided by the manufacturer, and column end plugs should be fitted to prevent solvent evaporation and drying out of the packing bed.

• Korean Journal of Food Science and Technology
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• v.29 no.4
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• pp.648-656
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• 1997

Thermal treatment of soymilk residue was carried out at 140, 150, and $160^{\circ}C$ for possible use as a raw material for dietary fiber, and some physicochemical properties of the extracts were investigated. Soluble dietary fiber(SDF) content of the extracts prepared under optimal conditions was more than 30% suggesting the conversion of insoluble dietary fiber to SDF. The main sugar components of the extracts were glucose, galactose, and arabinose. Analysis of the moelcular weight distribution by high performance size exclusion chromatography revealed that the proportion of high molecular weight fraction decreased and that of middle-sized polymer increased as the extraction temperature increased. The viscosity of aqueous solution of the extracts decreased with an increase in extraction temperature, but showed no trend as pH changed. The solubility increased with extraction temperature showing the highest at $160^{\circ}C$. The extract at $140^{\circ}C$ had the biggest calcium-binding capacity, which correlated with the changes in viscosity.

• Journal of Korea Technical Association of The Pulp and Paper Industry
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• v.43 no.5
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• pp.60-67
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• 2011

Hemicelluloses pre-extracted from Korean mixed wood chip were investigated as a wet-end additive. Hemicelluloses dissolved in hot water and alkali solution were isolated by ethyl alcohol precipitation from pre-extractives. They showed molecular weight of 9,000 ~ 27,000 g/mol as revealed by size exclusion chromatography. The reduction of molecular weight through hot water extraction was caused by autohydrolysis. Chemical composition of the hemicelluloses were analyzed with high-performance liquid chromatography and UV-Vis spectroscopy. As the surface charge of isolated hemicelluloses were negative, the adsorption of hemicelluloses onto softwood unbleached kraft pulp fiber was promoted by poly-DADMAC. The physical properties of handsheets increased as the molecular weight of hemicellulose increased. On the other hands, the optical property decreased with hemicellulose adsorption.

• Journal of Applied Biological Chemistry
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• v.59 no.2
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• pp.149-154
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• 2016

An endophytic bacterial strain was isolated from kimchi, a Korean traditional fermented Brassica campestris and identified as Bacillus subtilis MJMP2 based on the 16S rRNA sequence. This strain showed strong antagonistic activity against Xanthomonas oryzae pv. oryzae (Xoo) KACC10331, the pathogen of bacterial rice blight disease, as well as activity against some other rice phytopathogenic fungi. The active compound was purified through size-exclusion chromatography and preparative High-performance liquid chromatography. The molecular weight was determined as m/z 1043 by mass spectroscopy, which is identical to that of iturin A. Furthermore, a crude extract from the culture supernatant of Bacillus subtilis MJMP2 showed inhibitory activity against rice blight disease in both a rice leaf explant assay and a pot assay. The crude extract also enhanced the length of roots of Arabidopsis thaliana. These results suggest that the strain Bacillus subtilis MJMP2 could be used as a biological agent to control rice blight disease.

• Korean Journal of Food Science and Technology
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• v.31 no.3
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• pp.600-605
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• 1999

Methyl linoleate was oxidized at 60, 90, 120 and $150^{\circ}C$, respectively, with sparging oxygen for different periods of time. On the basis of the peroxide values determined at four temperatures, four heating times were chosen for the analysis of physicochemical parameters, such as peroxide value, total oxidation products, polymer content, viscosity, refractive index and characteristics of thermal degradation by DSC (Differential Scanning Calorimeter). The content of peroxide linkage (C-O-O-C) polymer and ether or carbon to carbon linkage (C-O-C/C-C) polymer were analyzed by High Performance Size Exclusion Chromatography (HPSEC). The polymer formed at four temperatures was qualitatively identified as dimer. The polymer with peroxide linkage (C-O-O-C) were detected from methyl linoleate oxidized at $60^{\circ}C\;and\;90^{\circ}C$, but they were not detected from methyl linoleate oxidized at $120^{\circ}C\;and\;150^{\circ}C$. The enthalpy changes increased as peroxide value increased whereas maximum degradation temperature decreased. The highest correlation coefficients were obtained between maximum degradation temperature $(T_m)$, exothermic enthalpy changes and peroxide value, peroxide linkage (C-O-O-C) polymer content.

• BMB Reports
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• v.39 no.5
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• pp.546-555
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• 2006

An acidic polysaccharide fraction that had high anti-complementary activity was isolated from the stems of Grey Mangrove in 0.15% yield. The final fractions was designated HAM-3-IIb-II. The polysaccharide fraction appeared to be homogenous by high performance size exclusion chromatography with an estimated molecular weight of 105 kDa. The isolated polysaccharide is more effective than polysaccharide K (PSK) in its anti-complementary activity at 58 ${\mu}g/ml$ of PSK and 23 ${\mu}g/ml$ of HAM-3-IIb-II that inhibit 50% of complement activity in the complement fixation assay. Structural studies indicated that HAM-3-IIb-II was rich in galacturonic acid along with arabinose, galactose and rhamnose, characterizing a pectin-type polysaccharide, which was also confirmed by FT-IR spectrum. The presence of rich neutral sugar side chains of arabinogalactans may have contributed to the expression of high activity. Traditionally, this mangrove plant is used for medicinal purposes and it appears to have some scientific applications.

• Journal of Microbiology and Biotechnology
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• v.19 no.10
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• pp.1201-1205
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• 2009

Corn starches with different amylose contents were enzymatically modified using Thermus aquaticus 4-$\alpha$-glucanotransferase ($TA{\alpha}GTase$). Upon the enzyme treatment, the chain-length distributions of isoamylolytically debranched products became broader [degree of polymerization (DP): 3-40] than those of untreated corn starches. In addition, a variety of cycloamyloses (CAs) with different sizes were formed by the glucanotransfer activity of $TA{\alpha}GTase$. CAs with DP 5-40 were detectable in all of the $TA{\alpha}GTase$-treated corn starches. From the results of high-performance anion-exchange chromatography and high-performance size-exclusion chromatography analyses, it was suggested that the amount of CAs produced by the enzyme treatment increased as the amylose content of the starches increased. Thus, we concluded that the extent of modification of starch molecules was enhanced in proportion to amylose content by the transfer activity of $TA{\alpha}GTase$. This finding could be useful for developing an efficient process of CA production using this enzyme.

• Environmental Engineering Research
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• v.12 no.5
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• pp.224-230
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• 2007

The objectives of this study were to evaluate the change of molecular weight (MW) profiles in natural organic matter (NOM) through various treatment processes (coagulation, granular activated carbon (GAC), and ozonation) using high performance size exclusion chromatography based on ultraviolet absorbance and dissolved organic detection (HPSEC-UVA-DOC). In addition, relationships between MW profiles and disinfection by-production (DBP) formation were evaluated. Each treatment process results in significant different effects on NOM profiles. Coagulation is effective to remove high molecular weight NOM, while GAC is effective to remove low molecular weight NOM. Ozonation removes only a small portion of NOM, while it induces a significant reduction of UV absorbance due to breakdown of the aromatic groups. All treated waters are chlorinated, and chlorination DBPs such as trihalomethanes (THMs) and haloacetic acids (HAAs) are measured under formation potential conditions. Both THM and HAA formation potentials were significantly reduced through the coagulation process. GAC was more effective to reduce THM formation compared to HAA formation reduction, while ozonation showed significant HAA reduction compared to THM reduction.

• Journal of Soil and Groundwater Environment
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• v.20 no.7
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• pp.70-79
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• 2015

A sample of soil humic acid (HA) was divided by ultrafiltration (UF) into five fractions of different molecular size (UF₁: > 300, UF₂: 100~300, UF₃: 30~100, UF₄: 10~30, UF₅: 1~10 kilodaltons). Apparent average molecular weight (M_w) of the HA fractions were measured using high performance size exclusion chromatography (HPSEC), and the chemical and structural properties of the five HA fractions were characterized by elemental compositions (H/C, O/C and w ((2O + 3NH)/ C)) and ultraviolet-visible absorption ratios (SUVA, A_4/6). The organic carbon normalized-sorption coefficients (Koc) for the binding of phenanthrene to the HA fractions were determined by fluorescence quenching and relationship between the sorption coefficients and structural characteristics of the HA fractions were investigated. The elemental analysis and UV-vis spectral data indicated that the HA fractions with higher molecular weights have grater aliphatic character and lower contents of oxygen, while the HA fractions with lower molecular size have greater aromatic character and molecular polarity that correspond to greater SUVA and internal oxidation values (w). The log Koc values (L/kg C) were gradual increased from 4.45 for UF₅ to 4.87 for UF1. The correlation study between the structural descriptors of the HA fractions and log Koc values of phenanthrene show that the magnitude of Koc values positively correlated with $M_w$ and H/C, while negatively correlated with the independent descriptors of the O/C, w, SUVA and A_4/6.