• KSBB Journal
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• v.14 no.6
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• pp.651-654
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• 1999

Using recombinant human growth hormone as a model protein, we carried out unfolding by adding a denaturant such as urea, guanidine HCl, or SDS followed by refolding by dilution and dialysis. The objectives were to monitor the structural changes during in vitro refolding process and, based on the results, to develop a quantitative method of refolding progress assessment. The changes in surface hydrophobicity were measured by fluorescence tagging of 1-anilinonaphthalene-8-sulfonate(1,8-ANS) to the hydrophobic portions, and those in the secondary structure were monitored by using far UV-CD(circular dichroism) spectroscopy. Also, we used RP-HPLC to separate and quantify the folded and unfolded proteins to correlate the result with the structure analysis. Our results indicate the surface hydrophobicity are well correlated with the formations of the secondary structure, primarily ${\alpha}$-helices, as well as the disulfide bridges. We expect this monitoring technique can be applied in industrial fields as a means to quantitatively assess the progress of in-vitro refolding of recombinant proteins.

• Korean Journal of Food Science and Technology
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• v.35 no.3
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• pp.503-509
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• 2003

For the development of the anti-obesity natural drug, the inhibitor of adipocyte differentiation was screened from Korean traditional medicinal plants. Phellodendri Cortex was selected as a candidate of adipocyte differentiation inhibitor. An inhibitory compound PC-4 was purified from the methanol (MeOH) extract of Phellodendri Cortex using silica gel and ODS RP-18 column chromatography and HPLC. PC-4 was obtained as yellow powder; UV ${\lambda}_{max}$ (MeOH): 230, 260, 340 and 430 nm. The chemical structure of PC-4 was determined as an isoquionoline alkaloid, berberine, on the basis of various NMR experiments including $^1H-\;and\;^{13}C-NMR$. The PC-4 inhibited the differentiation of preadipocyte NIH-3T3 L1 cells at a concentration of $1\;{\mu}g/mL$.

• Journal of the Korean Applied Science and Technology
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• v.38 no.6
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• pp.1568-1576
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• 2021

Here, we conducted the study on the possibility of using FK-13, a short analog of human-derived antibacterial peptide LL-37, as a cosmetic preservative to discover a natural cosmetic preservative that is safe for human body. For the purpose, FK-13 composed of 13 amino acids was synthesized by solid-phase peptide synthesis, and purified using reversed phase-high performance liquid chromatography (RP-HPLC). The purity and molecular weight were confirmed by liquid chromatography-mass spectrometry (LC-MS) analysis. FK-13 showed high antimicrobial activity on the three gram-positive bacteria (Staphylococcus aureus, Bacillus subtilis, and Staphylococcus epidermidis), the three gram-negative bacteria (Escherichia coli, Salmonella typhimurium, and Pseudomonas aeruginosa), and also even the fungus Candida glabrata. FK-13 had a broad spectrum of antibacterial activity, showing a suitability as a cosmetic preservative. In addition, FK-13 showed high thermostability and higher antibacterial activity in a comparative test with existing natural herbal cosmetic and chemical preservatives. Therefore, as FK-13 is a safe material and has high antibacterial activity at a low concentration, it is likely to be applied as a peptide natural cosmetic preservative that can replace existing chemical preservatives.

• Journal of Life Science
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• v.34 no.8
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• pp.576-587
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• 2024

In this study, diverse bacterial strains were isolated from fermented foods to screen those with antibacterial activity. Among them, one strain, identified as Bacillus vallismortis MCBL 1012 through 16S rRNA gene sequence analysis, was selected for its bacteriocin production. The culture supernatant of B. vallismortis MCBL 1012 showed antibacterial activity, mainly against Gram-positive bacteria. Scanning electron microscopy (SEM) revealed that bacteriocin treatment led to cellular content leakages in Listeria monocytogenes KCCM 40307, Enterococcus faecium KCCM 12118, and Streptococcus mutans KCTC 3065. PCR analysis confirmed B. vallismortis MCBL 1012 harbored subtilosin A gene (sbo A). Antibacterial activity was decreased by proteolytic enzymes like proteinase K, subtilisin A, and α-chymotrypsin. The bacteriocin demonstrated stability at 40℃ and 60℃ for 120 min, and up to 80℃ for 60 min, with rapid activity loss at 100℃. It retained full antibacterial activity within a pH range of 4.0 to 8.0 and was not affected by up to 100% organic solvents like ethanol, methanol, acetonitrile, and tetrahydrofuran. Nevertheless, activity decreased with more than 40% isopropanol and 80% acetone. Most tested inorganic salts and detergents had no effect on antibacterial activity except, CuSO₄ and NiSO₄ at specified concentrations. The bacteriocin exerted its antibacterial effect through bactericidal action against L. monocytogenes KCCM 40307. The bacteriocin was purified by ammonium sulfate precipitation, DEAE anion exchange chromatography, and RP-HPLC. The purification resulted in a final yield of 0.03% and a 283.7-fold increase in specific activity. MALDI-TOF MS analysis determined the exact molecular weight of purified bacteriocin to be 3,326.1 Da.

• Korean Chemical Engineering Research
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• v.53 no.6
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• pp.723-729
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• 2015

The moment analysis of cyclic adenosine monophosphate (cAMP) was performed using chromatograms that were obtained with the pulse input method from an octadecyl silica (ODS) high-performance liquid chromatography (HPLC) column. The general rate (GR) model was employed to calculate the first absolute moment and the second central moment. Three important coefficients for moment analysis, which are molecular diffusivity ($D_m$), external mass transfer coefficient ($k_f$), and intra-particle diffusivity ($D_e$), were estimated by the Wilke-Chang equation, Wilson-Geankoplis equation, and comparing van Deemter equation to theoretical plate number equation, respectively. Experiments were conducted by various conditions of flow rates, methanol volume ratio of the mobile phase, and solute concentration. After the moment analysis, results were organized by van Deemter plots. Also van Deemter coefficients were compared each other to effect $H_{ax}$, $H_f$, and $H_d$ on height equivalent to a theoretical plate (HETP, $H_{total}$). The value of intraparticle diffusion ($H_d$) was the primary factor which makes for HETP whereas external mass transfer ($H_f$) was disregardable factor.

• Korean Journal of Food Science and Technology
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• v.41 no.3
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• pp.231-237
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• 2009

Trans free margarine stock (TFMS) was produced by lipase-catalyzed synthesis of fully hydrogenated soybean oil (FHSBO), avocado oil (AO) and palm oil (PO). A blend of FHSBO, AO, and PO with a 1:5:4 (30:150:120 g, respectively) ratio was interesterified with lipozyme RM IM(from Rhizomucor miehei) in a 1 L-batch type reactor at 65 for 12 hr, and the physicochemical and melting properties of TFMS were compared with commercial margarine. The solid fat content (%) of the TFMS was analyzed at 25, 30, and $35^{\circ}C$, respectively, while its melting point was $37.8^{\circ}C$. The trans fatty acid content of the TFMS was below 0.1%. It also had acid, saponification, and iodine values of 0.4, 173.9, and 58.6, respectively. In HPLC chromatograms of the TFMS, newly synthesized peaks of triacylglycerol molecules were observed by using reverse-phase HPLC with evaporative light-scattering detection. Normal-phase HPLC with UV detection was used to quantify tocopherols in the TFMS, indicating that its ${\alpha}-$, ${\gamma}-$ and ${\delta}$-tocopherol contents were 5.7, 2.1, and 1.7 mg/100 g, respectively.

• Analytical Science and Technology
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• v.29 no.4
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• pp.202-208
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• 2016

본 연구에서는 사료 첨가제로 이용되고 있는 유기산 7종(formic acid, malic acid, lactic acid, acetic acid, citric acid, fumaric acid, propionic acid)의 동시분석법 개발을 위한 연구를 실시하였다. 7종의 화합물은 표준물질의 Retention time과 UV spectra를 통해 구별하였고, 분석법 검증은 직선성, 민감성, 선택성, 정확성, 정밀성을 통하여 검증하였다. 그 결과로 LOD와 LOQ의 범위가 각각 43~26,755 μg/kg, 12-8,026 μg/kg으로 설정하였고, 평균 회수율이 79.3~95.2%로 우수하게 보였으며, intra-day, inter-day에 대한 전반적인 상대 표준 편차(%RSD)는 3.2% 미만으로 나타났다. 이와 같이 검증된 자료를 통해 유기산의 동시분석에 대한 직선성, 민감성, 선택성, 정확성 및 정밀성을 확인하였고, 높은 수준을 나타냄을 알 수 있었다. 이를 바탕으로 유기산이 검출되는 단미사료 46 가지를 분석에 적용하여 진행하였고, 정량과 동시분석 검출을 위한 방법은 RP-HPLC/UV 검출기를 이용하여 성공적으로 개발되었다. 따라서 본 연구결과를 바탕으로 하여 사료 중의 유기산의 분석이 신속하고 정확해졌을 뿐 아니라, 다른 종류의 사료 또한 이를 적용하여 효율적으로 이용할 수 있을 것으로 판단된다.

• Applied Biological Chemistry
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• v.50 no.4
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• pp.316-320
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• 2007

Three phytotoxic compounds were isolated from the culture broth of Chaetomium sp. through silica gel column chromatography and HPLC (RP-18). Their chemical structures were elucidated as chaetoglobosin F, chaetoglobosin C and chaetoglobosin E on the basis of instrumental analyses such as $^1H-NMR,\;^{13}C-NMR$, and HMQC. They inhibited the root growth of barnyard grass with the $IC_{50}$ values of 66, 65, and $67{\mu}g/ml$, respectively.

• Korean Journal of Food Science and Technology
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• v.36 no.4
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• pp.537-541
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• 2004

Based on reaction time and substrate molar ratio, structured lipid (SL-corn) was produced at 1:2:2(corn oil/capric acid/CLA) and 4% immobilized lipase from Rhizomucor miehei (RM IM). Reaction was carried out for 24 hr at $55^{\circ}C$ in 1-L stirred-batch reactor. After reaction, 13.3 mol% capric acid and 8.9%, CLA were incorporated into corn oil. Iodine and saponification values of SL-corn were 68 and 202, respectively. Tocopherol content decreased after reaction (about 39%). SL-corn showed more yellowish color than corn oil (p<0.05). Reversed-phase HPLC indicated triacylglycerol species containing capric acid in SL-corn resulted in faster crystallization than that of corn oil.

• Korean Chemical Engineering Research
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• v.47 no.2
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• pp.258-261
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• 2009

In this work, we the ettect on extraction amounts and general composition content of phytoestrogen genistein and formononetin extracted from Sophoraflavescens Aiton by various ultrasonic waves(35, 72, and 170 KHz) and extraction time(30, and 60 min) were compared using extraction solvent water 100%. The pretreatment step was composed of ultrasonic waves extraction, filtration, concentration, and membrane filtration. The extracted sample was analyzed by reversed-phase high performance liquid chromatography(RP-HPLC). And the mobile phase applied was linearly changed with A/B of 80/20~65/35 vol% for 60 min(A water/acetic acid, 99.9/0.1 vol%, B acetonitrile/acetic acid, 99.9/0.1 vol%). The experimental results, general composition carbohydrate(0.255 to 0.413%) excepts, other ingredients was confirmed almost similarly. Also, The highest yield of extraction amount 3.17g was obtained by ultrasonic waves with a frequency of 170 KHz and an extraction time of 60 min. This work offers would be useful for chemical and biological studies of natural plants and its products.