• Journal of Marine Bioscience and Biotechnology
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• v.2 no.3
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• pp.142-148
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• 2007

An agar-degrading marine bacterium, strain AS-3 was isolated from the seawater. The strain AS-3 was identified as Algibacter lectus AS-3 by 16S rDNA sequence. The optimum medium for agarase activity of the isolated strain was determined to be marine medium, marine broth 2216 containing 0.1% agar as carbon source. An extracellular agarase was purified 6.9-fold from the culture supernatant by ammonium sulfate precipitation, ion exchange chromatography and gel filtration methods. The optimum pH and temperature for this enzyme were 7.0 and $40-50^{\circ}C$, respectively. Antioxidative activity of the strain AS-3 was 62.4% in the supernatant cultured for 12 h.

• The Korean Journal of Food And Nutrition
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• v.9 no.4
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• pp.404-408
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• 1996

One of the major objectives of the food industry is the enrichment of the functional properties and nutritional value of soybean protein. To attain this goal, an expression system of cDNA encoding native and protein-engineered soybean proteins in a microorganism must be developed and the function then ability of self-assembly and the functionalities of the expressed proteins should be evaluated before the modified genes are transfered to soybean plants. The pro-$\beta$-conglycinin synthesized in E. coli BL21(DE3) comprised approximately 20% of the total bacterial proteins and the expressed protein are formed soluble and trimer such as native protein in E. coli cells. The highly expressed protein was purified to homogeneity by salt precipitation with 20~40$ Ammonium sulfate ion-exchange chromatography with Q-Sepharose and hydrophobic column chromatography with Butyltoyopearl. Therefore, we concluded that the high-level expression system of $\beta$-conglycinin cDNA was established and a relatively simple and rapid method for purifying pro-$\beta$-conglicinin was also developed.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.41 no.4
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• pp.456-464
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• 1996

Fructans are the major vegetative storage carbohydrate in wheat(Triticum aestivum L.). The depolymerization of fructans occurs by the sequential removal of terminal fructosyl residues by a specific fructan exohydrolase(FEH). The objective of this study was to isolate and characterize this enzyme in wheat. From stems and sheaths of field-grown wheat(cv. Clark), FEH was purified 356-fold using salt precipitation and a series of chromatographic procedures including size exclusion, anion exchange, and affinity chromatography. FEH had a molecular weight of 63.7 kD and an optima at pH 5.5 and 3$0^{\circ}C$. The $K_{m}$s for $\beta$(2 longrightarrow1) linked oligofructans varied, from 10 to 37mM, with the lowest $K_{m}$ for tetrasaccharide. The $V_{max}$ increased as degree of polymerization (DP) increased. Wheat FEH hydrolyzed only, $\beta$(2 longrightarrow1) linked fructans but not, $\beta$(2 longrightarrow6) linked timothy fructan or sucrose. The role of this FEH in fructan metabolism in wheat is discussed.sed.

• Journal of Microbiology and Biotechnology
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• v.17 no.11
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• pp.1751-1757
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• 2007

Elastin-like polypeptides (ELPs) undergo a reversible inverse phase transition upon a change in temperature. This thermally triggered phase transition allows for a simple and rapid means of purifying a fusion protein. Recovery of ELPs-tagged fusion protein was easily achieved by aggregation, triggered either by raising temperature or by adding salt. In this study, levansucrase has been used as a model enzyme in the development of a simple one-step purification method using ELPs. The levansucrase gene cloned from Pseudomonas aurantiaca S-4380 was tagged with various sizes of ELPs to functionally express and optimize the purification of levansucrase. One of two ELPs, ELP[V-20] or ELP[V-40], was fused at the C-terminus of the levansucrase gene. A levansucrase-ELP fusion protein was expressed in Escherichia coli $DH5{\alpha}$ at $37^{\circ}C$ for 18 h. The molecular masses of levansucrase-ELP[V-20] and levansucrase-ELP[V-40] were determined as 56 kDa and 65 kDa, respectively. The phase transition of levansucrase-ELP[V-20] occurred at $20^{\circ}C$ in 50 mM Tris-Cl (pH 8) buffer with 3 M NaCl added, whereas the phase transition temperature ($T_t$) of levansucrase-ELP[V-40] was $17^{\circ}C$ with 2 M NaCl. Levansucrase was successfully purified using the phase transition characteristics of ELPs, with a recovery yield of higher than 80%, as verified by SDS-PAGE. The specific activity was measured spectrophotometrically to be 173 U/mg and 171 U/mg for levansucrase-ELP[V-20] and levansucrase-ELP[V-40], respectively, implying that the ELP-tagging system provides an efficient one-step separation method for protein purification.

• Microbiology and Biotechnology Letters
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• v.41 no.4
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• pp.462-468
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• 2013

Jeotgal is a generic term given to the high-salt-fermented seafood of Korea. This study aimed at developing an overview of the bacterial community present in Ojingeo-jeotgal, a highly consumed type of jeotgal, which is made with squid. Bacteria were isolated and purified from two samples on six different kinds of media and identified by 16S rRNA gene sequence analysis. Among the 121 total isolates, the most dominant genus was Bacillus, followed by coagulase-negative staphylococci (CNS) and lactic acid bacteria (LAB). CNS were detected in both samples, but LAB were observed in only a single sample. Six strains of Bacillus species inhibiting the growth of food pathogens, Staphylococcus aureus and Vibrio parahaemolyticus, were selected from the 121 isolates. These were found to inhibit the growth of both pathogens in addition to displaying proteolytic activities on media containing 6% NaCl and 2% skim milk.

• Journal of Microbiology and Biotechnology
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• v.19 no.1
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• pp.108-112
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• 2009

A simplified method for the purification of cholera toxin was developed. The 569B strain of Vibrio cholerae, a recognized hyper-producer of cholera toxin, was propagated in a bioreactor under conditions that promote the production of the toxin. The toxin was separated from the bacterial cells using 0.2-${\mu}m$ crossflow microfiltration, the clarified toxin was passed through the membrane into the permeate, and the bacterial cells were retained in the retentate. The 0.2-${\mu}m$ permeate was then concentrated 3-fold and diafiltered against 10 mM phosphate buffer, pH 7.6, using 30-kDa crossflow ultrafiltration. The concentrated toxin was loaded onto a cation exchange column, the toxin was bound to the column, and most of the impurities were passed unimpeded through the column. The toxin was eluted with a salt gradient of phosphate buffer, pH 7.0, containing 1.0 M NaCl. The peak containing the toxin was assayed for cholera toxin and protein and the purity was determined to be 92%. The toxin peak had a low endotoxin level of $3.1\;EU/{\mu}g$ of toxin. The purified toxin was used to prepare antiserum against whole toxin, which was used in a $G_{M1}$ ganglioside-binding ELISA to determine residual levels of toxin in an oral inactivated whole-cell cholera vaccine. The $G_{M1}$ ganglioside-binding ELISA was shown to be very sensitive and capable of detecting as little as 1 ng/ml of cholera toxin.

• Journal of Applied Biological Chemistry
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• v.60 no.3
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• pp.257-263
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• 2017

A cellulolytic strain Y2 was isolated from soil obtained in the Canadian Alpine region. The isolate was identified as Paenibacillus sp. Y2 by 16S rRNA sequencing. When grown in LB medium supplemented with carboxymethyl-cellulose (CMC), CMCase production increased to 122.0% of that observed in LB without CMC. Culture supernatant was concentrated by ultrafiltration and 80% ammonium sulfate precipitates were separated by Hi-Trap Q and CHT-II chromatography. The purified enzyme (EG-PY2) showed a homogeneous single band and the molecular mass was estimated to be 38 kDa by SDS-PAGE. Optimum pH and temperature of the enzyme were 4.5 and $30^{\circ}C$, respectively. The half-life of enzyme activity at 50 was 140.7 min, but the enzyme was drastically inactivated within 5 min at $55^{\circ}C$. The enzyme was highly activated to 135.7 and 126.7% by 5.0 mM of $Cu^{2+}$ or $Mg^{2+}$ ions, respectively, and moderately activated by $Ba^{2+}$ and $Ca^{2+}$ ions, whereas it was inhibited to 76.8% by $Fe^{2+}$, and to ${\leq}50%$ by $Mn^{2+}$, $Co^{2+}$, $Zn^{2+}$, and EDTA. The enzyme was activated to 211.5% in the presence of 0.5 M of NaCl and greatly tolerant to 3.15M of NaCl. The enzyme showed 2.98 times higher ${\beta}$-glucanase activity than CMCase activity. Based on these results, it can be concluded that EG-PY2 is an acidophilic, cold-active, and halotolerant endoglucanase. The authors suggest it is considered to be useful for various industrial applications, such as, fruit juice clarification, acidic deinking processes, high-salt food processing, textile and pulp industries, and for biofuel production from seaweeds.

• Food Science and Preservation
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• v.19 no.2
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• pp.229-234
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• 2012

The antioxidant substance in black onion was identified. The assays that used 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzthiazo line-6-sulphonic acid) ($ABTS^+$) radicals showed that the ethyl acetate fraction of black onion methanol extract had a higher level of radical-scavenging activity than the other fractions. Two antioxidative compounds were purified and isolated from the ethyl acetate fraction via column chromatographies of silica gel and Sephadex LH-20 using the guided DPPH radical-scavenging assay. The isolated compounds were identified as 3,4-dihydroxybenzoic acid (1) and quercetin (2) based on mass spectrometry and nuclear magnetic resonance. The isolated compounds showed a high level of DPPH and ABTS+ radical-scavenging activity. Compound 2 had a higher level of radical-scavenging activity than 1.

• Microbiology and Biotechnology Letters
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• v.39 no.3
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• pp.218-223
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• 2011

To obtain eicosapentaenoic acid (EPA)-producing bacteria, some 600 strains of bacteria were isolated from Antarctic sediment and marine organisms during the summer expedition of 1999-2000 and 7 EPA-producing bacteria were obtained through screening with TLC and GC. A strain designated as L93 showed the highest EPA production, which was gram-negative, rod-shaped bacterium. L93 strain was identified as Shewanella sp., from the sequence analysis of 16S rDNA. Optimal conditions temperature and pH for the growth and EPA production were about $4^{\circ}C$ and pH 7. In addition, its production was optimized by 50%(w/v) sea salt. We establish the optimal production system to produce about 320 mg per liter by using this optimal EPA production conditions. EPA-methyl ester was purified from cultured L93 strain to a purity of higher than 97% and typical purification yield is greater than 72% of the input amount via urea complexation and HPLC.

• Journal of Microbiology and Biotechnology
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• v.24 no.1
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• pp.87-96
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• 2014

In the present study, a yeast species isolated from CETP, Vellore, Tamilnadu was identified as Cryptococcus sp. VITGBN2 based on molecular techniques and was found to be a potent producer of acidic diacetate sophorolipid in mineral salt media containing vegetable oil as additional carbon source. The chemical structure of the purified biosurfactant was identified as acidic diacetate sophorolipid through GC-MS analysis. This sophorolipid was used as a stabilizer for synthesis of zinc oxide nanoparticles (ZON). The formation of biofunctionalized ZON was characterized using UV-visible spectroscopy, XRD, scanning electron microscopy (SEM), and Fourier transform infrared spectroscopy. The antimicrobial activities of naked ZON and sophorolipid functionalized ZON were tested based on the diameter of inhibition zone in agar well diffusion assay, microbial growth rate determination, protein leakage analysis, and lactate dehydrogenase assay. Bacterial pathogen Salmonella enterica and fungal pathogen Candida albicans showed more sensitivity to sophorolipid biofunctionalized ZON compared with naked ZON. Among the two pathogens, S. enterica showed higher sensitivity towards sophorolipid biofunctionalized ZON. SEM analysis showed that cell damage occurred through cell elongation in the case of S. enterica, whereas cell rupture was found to occur predominantly in the case of C. albicans. This is the first report on the dual role of yeast-mediated sophorolipid used as a biostabilizer for ZON synthesis as well as a novel functionalizing agent showing antimicrobial property.