• Journal of Biosystems Engineering
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• 제33권4호
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• pp.276-281
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• 2008

A flow injection analysis method (FIA), which analyzes sample conditions after injecting a sample and reagents into a continuous stream, are recognized as the most adequate analyzing method according to the increase of sampling frequency, the decrease of measuring time and the diversity of measuring targets. Specially, the FIA is considered to be used effectively for the control of a fermentation process to produce fermentation food and useful microbial production by activation of a fermentation industry for development of biological materials. In this study, a flow injection analysis sensor unit was developed for on-line monitoring of the fermentation process. The performance was verified by on-line measuring the concentration of glucose of the fermentation process of wine. The glucose concentrations of the samples were measured every 12 hours during the whole fermentation process and compared with those by a HPLC. The concentration relative errors of glucose on the fermentation process of wine showed below 30% within 72 hours and over 50% after the 72 hours. The sensor unit had potential to on-line monitoring of the fermentation process but some problems to overcome for an commercial application.

• 한국환경과학회지
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• 제7권2호
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• pp.203-208
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• 1998

A simple on-line measurement system consisting of a conventional peristaltic pump, a HPLC-type heater, and a flow-through spectrophotometer is introduced for the determination of chemical oxygen demand(CODI. The system was configured such that the reaction mixture in the highly concentrated surffuric acrid medium flowing through the PTFE reaction tubing was heated at 150℃ and the absorbance of dichromate was continuously moutored at 445 m. The same addation principle as in the standard procedure was employed akcept the use of CoSO4 as a new effective catalyst. To test the system, potassium hydrogen phthalate was selected as a COD standard material. With suitably optimized reaction condition, the applicable concentration range depends on the concentration of potassium dichromate in the oxidizing reagent. With 2.0×10-3 M and 5.0×10-4M dichromate, the linear dynamic range was observed up to 400 ppm and 100 ppm, respectively. The standards in the Unear ranges were shown to be completely oxidized, which was confirmed with sodium oxalate or Mohr's salt. In all cases, the typical reproduclbility for betweenruns was 2% or less. The proposed measurement system provides the valuable in- formation for the further development of automated analysis system based on the present standard procedure.

• KSBB Journal
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• 제32권1호
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• pp.35-45
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• 2017

In this work the optical fiber glucose and lactate biosensors were developed by using fluorescent dye and enzyme immobilized on the end tip of an optical fiber. 3-Glycidyloxypropyl)methyldiethoxysilane (GPTMS), (3-Aminopropyl) trimethoxysilane (APTMS) and Methyltrimethoxysilane (MTMS) were used to immobilize glucose oxidase (GOD), lactate oxidase (LOD) and ruthenium(II) complex (tris(4,7-diphenyl-1,10-phenanthroline) ruthenium(II), $Ru(dpp)_3^{2+}$) as oxygen sensitive fluorescent dye. MTMS sol-gel was an excellent supporting material for the immobilization of $Ru(dpp)_3^{2+}$, GOD, and LOD on the optical fiber. Storage stability of the optical fiber glucose sensor was kept constant over 20 days, while the optical fiber lactate sensor had constant storage stability over 17 days. The optical fiber glucose and lactate biosensors also maintained good operational stability for 20 hours and 14 hours, respectively. The activities of the immobilized enzymes were most excellent at pH 7 and at $25^{\circ}C$. On-line monitoring of glucose and lactate in a simulated process was performed with the optical fiber glucose and lactate biosensors. On-line monitoring results were agreed with those of off-line data measured with high performance liquid chromatography (HPLC).

• 한국식품영양과학회지
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• 제30권4호
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• pp.732-738
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• 2001

두충잎에서 인간대장암에세포 HCT-116에 세포증식 억제효과를 나타내는 성분을 silica gel column chromatography TLC로 분리하고 HPLC로 정제한 다음 UV-visual spectro-photometer에 의한 흡수 spectra 특성 HPLC에 의한 retention time과 분리패턴 및 FAB/MS로 분자량을 잠정적으로 동정한 결과 TLC 상에서 분리된 Rf 0.19 band 의 화합물은 chlorophyll a에서 $Mg^{2+}$이 제거된 pheophytin a로 Rf 0.25 band의 화합물은 pyropheophytin a로 확인되었다. 동정된 화합물들의 암세포 억제작용을 확인하기 위해 두충잎의 petroleum ether extract column chromatography로 분리한 분획물 및 TLC에서 분리된 각 band들의 cytotoxic activity을 in vitro에서 HCT-116 cancer cell을 사용하여 측정한 결과 모든 시료에서 암세포 증식억제 현상을 보였으며 특히 Rf 0.19와 Rf 0.25 band에서 분리된 화합물들에서 강하게 나타났다. 이 결과로 볼 때 두충잎에서 분리한 클로로필 유도체는 인간대장암세포인 HCT-116 cell에 대해 강한 항암작용이 있는 것으로 생각된다.

• Journal of Ginseng Research
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• 제37권3호
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• pp.332-340
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• 2013

In this study, gamma rays were used to irradiate embryogenic calli induced from cotyledon explants of Panax ginseng Meyer. After the embryogenic calli were irradiated, they were transferred to adventitious roots using an induction medium; next, mutated adventitious root (MAR) lines with a high frequency of adventitious root formations were selected. Two MAR lines (MAR 5-2 and MAR 5-9) from the calli treated with 50 Gy of gamma rays were cultured on an $NH_4NO_3$-free Murashige and Skoog medium with indole-3-butyric acid 3 mg/L. The expression of genes related to ginsenoside biosynthesis was analyzed using reverse transcription polymerase chain reaction with RNA prepared from native ginseng (NG), non-irradiated adventitious root (NAR) and 2 MAR lines. The expression of the squalene epoxidase and dammarenediol synthase genes was increased in the MAR 5-2 line, whereas the phytosterol synthase was increased in the MAR 5-9 line. The content and pattern of major ginsenosides (Rb1, Rb2, Rc, Rd, Re, Rf, and Rg1) were analyzed in the NG, NAR, and 2 MAR lines (MAR 5-2 and MAR 5-9) using TLC and HPLC. In the TLC analysis, the ginsenoside patterns in the NG, NAR, and 2 MAR lines were similar; in contrast, the MAR 5-9 line showed strong bands of primary ginsenosides. In the HPLC analysis, compared with the NG, one new type of ginsenoside was observed in the NAR and 2 MAR lines, and another new type of ginsenoside was observed in the 2 MAR lines irradiated with gamma rays. The ginsenoside content of the MAR 5-9 line was significantly greater in comparison to the NG.

• 한국축산식품학회지
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• 제37권3호
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• pp.402-409
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• 2017

A novel peptide having free radical scavenging activity was separated, using an on-line high-performance liquid chromatography (HPLC) - ABTS screening method, from bovine skim milk fermented by Lactococcus lactis SL6 (KCTC 11865BP). It was further purified using reverse phase-HPLC (RP-HPLC) and sequenced by RP-HPLC-tandem mass spectrometry. The amino acid sequence of the identified peptide was determined to be Phe-Ser-Asp-Ile-Pro-Asn-Pro-Ile-Gly-Ser-Glu-Asn-Ser-Glu-Lys-Thr-Thr-Met-Pro-Leu-Trp (2,362 Da), which is corresponding to the C-terminal fragment of bovine ${\alpha}_{s1}$-casein (f179-199). The hydroxyl radicals scavenging activity ($IC_{50}$ $28.25{\pm}0.96{\mu}M$) of the peptide chemically synthesized based on the MS/MS data showed a slightly lower than that of the natural antioxidant Trolox ($IC_{50}$ $15.37{\pm}0.52{\mu}M$). Furthermore, derivatives of the antioxidant peptide were synthesized. The antioxidative activity of the derivatives whose all three proline residues replaced by alanine significantly decreased, whereas replacement of two proline residues in N-terminal region did not affect its antioxidative activity, indicating that $3^{rd}$ proline in C-terminal region is critical for the antioxidative activity of the peptide identified in this study. In addition, N-terminal region of the antioxidant peptide did not show its activity, whereas C-terminal region maintained antioxidative activity, suggesting that C-terminal region of the peptide is important for antioxidative activity.

• Food Science and Biotechnology
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• 제18권3호
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• pp.672-681
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• 2009

The aim of this study was to evaluate the antioxidant activities of enzymatic digests from citrus by-products (CBPs) prepared by high speed drying (HSD). HSD needs a short time (60 min) for drying and can be used in a commercial scale. Enzymatic digests were prepared from the CBPs using 6 enzymes such as aminoglucosidase (AMG), celluclast, pectinex, termamyl, ultraflo, and viscozyme. Antioxidant activities of AMG digest from CBPs were evaluated by different in vitro models such as 1,1-diphenyl-2-picrylhydrazyl (DPPH), hydroxyl, alkyl, $H_2O_2$ scavenging, metal chelating, lipid peroxidation, and comet assays, and exhibited strong activities. The antioxidant compounds were detected by an high performance liquid chromatography (HPLC) coupled on-line to an 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid ($ABTS^+$) scavenging detection system, and found that hesperidin was the key compound. Therefore, the results obtained in this study suggest that HSD is an effective method to transform wet CBPs into dried form, and CBPs are potential source of natural antioxidant.

• KSBB Journal
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• 제18권2호
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• pp.100-106
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• 2003

의학, 약학적으로 작용이 뛰어난 약리성분이 인체 내에서 상처 부위에 얼마나 빨리 도달하여 치료를 하는지를 알기 위하여 혈장 단백질과 약리성분의 결합력을 연구하는 새로운 연구 분야가 HPFA이다. 본 연구는 인체 내에 존재하는 혈장단백질과 항암제로서 알려져 있는 (S)-perillyl alcohol의 결합력과 결합 매개변수를 구하기 위하여 on-line frontal analysis HPLC system을 적용하였다. Develosil 100 Diol 5 (10 cm $\times$ 4.6 mm I.D.)의 HPFA 컬럼을 이용하였고, 이동상은 인산완충용액(pH = 7.4, I = 0.17)을 이용하였다. UV wavelength 205 nm에서 실험을 수행하였고, 주입 부피는 본 실험 조건인 혈장 단백질이 350 rM일 경우에 최대의 혈장 단백질과 결합되지 않은 약리성분의 농도를 갖게 되는 600 ${\mu}\ell$으로 정하였다. Scatchard analysis를 통한 연구 결과로 혈장 단백질인 HSA와 (S)-perillyl alcohol의 결합 매개변수(K)와 단위 HSA에 대한 S-POH의 결합 위치의 수(n)는 각각 K : 2.05 $\times$ $10^{6}$ [M$^{-1}$], n : 0.00428로 실험적으로 구하였다

• Biotechnology and Bioprocess Engineering:BBE
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• 제10권3호
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• pp.198-204
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• 2005

Seeds of neem were collected from different parts of India and analyzed for their azadirachtin content by High Performance Liquid Chromatography (HPLC). In order to assess the effects of genotypic and geographical variation on azadirachtin content in cell cultures, callus development was attempted in the seeds containing high and low concentration of azadirachtin. The concentration of azadirachtin in callus cultures was significantly affected by the explant source. Seed kernels with higher azadirachtin content produced higher azadirachtin content in callus cultures and lower azadirachtin content was seen in callus cultures produced from seed kernels with low azadirachtin content. The protocol for development of elite stock culture of Azadirachta indica was established with the objective of selecting a high azadirachtin-producing cell line. The highest azadirachtin-producing cell line was selected and the effects of different media and illumination conditions on growth and azadirachtin production were studied in shake flask suspension culture. Detailed batch growth kinetics was also established. These studies provided elite starter culture and associated protocols for cultivation of A. indica plant cell culture in the bioreactor.

• KSBB Journal
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• 제18권5호
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• pp.377-384
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• 2003

본 연구에서는 푸마르산 또는 숙신산을 생산/소비하는 생물공정에서 각 물질의 농도를 온라인 모니터링을 하기 위한 흐름주입분석기술 (FIA)을 개발하였다. 각 효소반응에 필요한 효소 (Fum/MDH, ICL/ICDH)를 VA-Epoxy Biosynth E3-carrier에 공유 결합에 의해 고정화하였고 소형 고정화 효소반응기를 흐름주입분석 장치에 도입하였다. 푸마르산-FIA 및 숙신산-FIA 시스템에서 고정화 효소 반응기의 특성 (예를 들면, 고정화 Fum/MDH 효소 반응기와 ICL/ICDH 효소 반응기는 최적 pH가 9.0이었으며 35$^{\circ}C$의 반응온도에서 최대 활성을 보임) 을 연구하였고 실제 생물공정에의 적용 가능성을 조사하기 위해 각 FIA 시스템의 성능을 조사하였다. 또한 모사공정과 실제 발효공정에서 푸마르산과 숙신산의 농도를 온라인 모니터링하여 오프라인 분석값과 비교하였고 잘 일치함을 볼 수 있었다.