• Title/Summary/Keyword: bioprocess

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Optimal Culture Conditions and XAD Resin on Tropane Alkaloid production in Scopolia parviflora Hairy Root Cultures (미치광이풀 모상근의 배양조건 구명 및 XAD Resin 처리에 의한 Tropane Alkaloid 생산)

  • 정희영;강민정;강영민;윤대진;박정동;정영관;최명석
    • KSBB Journal
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    • v.17 no.6
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    • pp.525-530
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    • 2002
  • The optimum culture conditions for tropane alkaloid production in hairy root cultures of Korea native Scopolia paviflora Nak. were investigated. Hairy root was induced from the rhizome of the mother plant on B5 medium containing 1.0 mg/L IBA. Among the culture media examined, 1/2 B5 medium was the best for tropane alkaloid production, whereas the growth of hairy root increased in SH medium. The best result on the growth of hairy root was obtained in 1.0 mg/L NAA, and tropane alkaloid production was obtained in plant growth regulator-free medium. Of the carbone sources tested, 3% sucrose promoted the growth of hairy root, whereas 5% sucrose increased tropane alkaloid production. Optimum inoculum densities for root growth and tropane alkaloid production were 0.5 g and 1 g, respectively. The addition of XAD resins (1 % w/v) to hairy root cultures led to increases in tropans alkaloid production, and the release of alkaloid into the medium and its adsorption by the resin accounted for about 50 to 80% of total production. It is concluded that optimized culture conditions and the addition of XAD resins could be used in the development of a bioprocess for tropane alkaloid production in hairy root cultures of S. paviflora Nak.

Optimal Production of Xylooligosaccharide by Using Recombinant Endoxylanase from Bacillus subtilis (Bacillus subtilis 유래 재조합 endoxylanase를 이용한 xylooligosaccharide의 최적 생산)

  • Kim, Yeon-Hee;Heo, Sun-Yeon;Kim, Mi-Jin;Lee, Jae-Hyung;Kim, Young-Man;Nam, Soo-Wan
    • Journal of Life Science
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    • v.18 no.1
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    • pp.52-57
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    • 2008
  • Xylan is a major hemicellulose component of the cell walls of monocots and hardwood, representing up to 30% of the dry weight of these plants. To efficiently hydrolyze xylan, the endoxylanase gene from Bacillus sp. was expressed in B. subtilis DB431 by introducing the plasmid pJHKJ4. The total activity of the recombinant endoxylanase reached about 857 unit/ml by batch fermentation of B. subtilis DB431/pJHKJ4 in LB maltose medium. The majority (>92%) of endoxylanase was efficiently secreted into the culture medium. The recombinant endoxylanase hydrolyzed more the birchwood xylan efficiently than the other xylans. When 4 % concentration of xylan was used, the highest production of xylooligosaccharide was observed, and xylobiose and xylotriose were the major products. Optimal amount of enzyme and reaction time for producing xylooligosaccharide were found to be 10 unit and 1 hr, respectively. In addition, the temperature of $40^{\circ}C{\sim}50^{\circ}C$ gave the highest production of xylooligosaccharide. Consequently, the optimized conditions for the production of xylooligosaccharide through the hydrolysis of xylan were determined as follows: 10 unit endoxylanase, $50^{\circ}C$, 4% birchwood xylan, 1 hr reaction.

Process Parameters on Quality Characteristics of Jacopever (Sebastes schlegeli Hilgendorf) under Treatment of Hydrostatic Pressure (고압처리 공정변수가 조피볼락의 초기 품질특성에 미치는 영향)

  • Kim, Min-Ji;Lee, Soo-Jeong;Kim, Chong-Tai
    • The Korean Journal of Food And Nutrition
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    • v.29 no.3
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    • pp.371-381
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    • 2016
  • The present study investigated the effects of processing parameters such as time (10, 20, 30, 40 min), pressure (25, 50, 75, 100 MPa), and the salinity of brine (0~10%(w/v)) on jacopever (Sebastes schlegeli Hilgendorf) in order to establish optimization of the three factors using a high hydrostatic pressure (HHP) machine. To do so, it analyzed the quality characteristics of volatile basic nitrogen (VBN), trimethylamine (TMA), total bacterial counts, dynamic viscoelasticities, and differential scanning calorimetry (DSC) properties. First, when the time increased to 40 mins, by 10 min intervals, the total bacterial counts in HHP groups under $25^{\circ}C$, 100 MPa, and 4%(w/v) brine were significantly decreased except for the first 10 min in comparison to the control group. In regards to DSC properties, the onset temperature ($T_O$) of the first endothermal curve was significantly reduced. Second, when the pressure level increased up to 100 MPa by 25 MPa increments, the total bacterial counts in the HHP samples significantly decreased for 20 min at 50 MPa or higher. As the pressure increased, G', G" and the slope of tan ${\delta}$ decreased (except for 50 MPa). Third, in regards to the salinities of brine, when the HHP processing was treated at 100 MPa, $25^{\circ}C$ for 20 min, the total bacterial counts of all the HHP groups significantly decreased in comparison to those of the control group. A significant difference was found in the enthalpy of the second endothermic curve in the 6~10%(w/v) (except 7%(w/v)) HHP groups. Therefore, the salinity of the immersion water under the HHP condition was appropriate when it was lower than 6%(w/v). The present study demonstrated that the optimum parameter condition according to/under the condition of the microbial inhibition and economic effects using an HHP would be the reaction time for 20 min, reaction pressure at 100 MPa, and the salinity of 4%(w/v) brine.

Antiproliferative Effect of Extracts from Corydalis heterocarpa on Human Cancer Cells (염주괴불주머니 (Corydalis heterocarpa) 추출물의 암세포 성장 억제 효과)

  • Kim, You-Ah;Lee, Jung-Im;Kong, Chang-Suk;Yea, Sung-Su;Seo, Young-Wan
    • KSBB Journal
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    • v.24 no.2
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    • pp.201-206
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    • 2009
  • Whole plants of Corydalis heterocarpa were extracted twice with $CH_2Cl_2$ and MeOH in turn. The combined crude extracts were concentrated in vacuo and then partitioned between $CH_2Cl_2$ and $H_2O$. The organic layer was fractionated with n-hexane and 85% aq. MeOH, and the aqueous fraction was also further fractionated with n-BuOH and $H_2O$, successively. Growth inhibition effects of crude extracts and their solvent fractions were evaluated in AGS, HT1080, U-937, MCF-7 and HT-29 human cancer cells using MTT assay. The inhibitory effects of solvent fractions were increased in a dose-dependent manner. Among these tested samples, 85% aq. MeOH fraction showed the most potent inhibitory effect on the growth of human cancer cells. These results suggest that active compounds having much stronger anticancer effect can be isolated from Corydalis heterocarpa.

Optimization for Effective Bioproduction of Natural (-)-1-Octen-3-ol by Lipoxygenase and Hydroperoxide Lyase from Agaricus bisporus (Agaricus bisporus의 Lipoxygenase와 Hydroperoxide Lyase를 이용한 천연 (-)-1-Octen-3-o1 생산 공정의 최적화)

  • Kim, Kyoung-Ju;Kim, Yong-Hwi
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.6
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    • pp.899-903
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    • 2005
  • One of the most important volatile aroma compounds responsible for mushroom flavor is 1-octen-3-ol. To meet the demand for natural mushroom flavor, a study was needed for the production of natural chiral specific (-)-1-octen-3-ol that has higher flavor intensity than synthetic chiral mixtures of (+), and (-)-1-octen-3-ol. The biosynthesis of (-)-1-octen-3-ol was achieved by an aerobic oxidation using lipoxygenase (LOX) and hydroperoxide lyase (HPOL) isolated from commercially available mushrooms in Korean market. Safflower oil from Uiseong, Gyeongsangbuk-do, that contains $\geq75\%$ of linoleic acid, was hydrolyzed using lipase. The recovered linoleic acid was biotransformed to stereo-specific 10-hydroperoxy linoleic acid by LOX. 10- hydroperoxy linoleic acid was further cleaved to (-)-1-octen-3-ol by HPOL. A commercial bioprocess for the production of (-)-1-octen-3-ol was developed using a 5-liter jar fermenter with fruiting bodies of Agaricus bisporus harvested from Buyeo, Chungcheongnam-do. The maximum production of (-)-1-octen-3-o1 was achieved at $4^{\circ}C$, pH 6.5 and 800 rpm yielding 748 mg/kg of mushroom.

GABA-enriched fermented Laminaria japonica improves cognitive impairment and neuroplasticity in scopolamine- and ethanol-induced dementia model mice

  • Reid, Storm N.S.;Ryu, Je-kwang;Kim, Yunsook;Jeon, Byeong Hwan
    • Nutrition Research and Practice
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    • v.12 no.3
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    • pp.199-207
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    • 2018
  • BACKGROUND/OBJECTIVES: Fermented Laminaria japonica (FL), a type sea tangle used as a functional food ingredient, has been reported to possess cognitive improving properties that may aid in the treatment of common neurodegenerative disorders, such as dementia. MATERIALS/METHODS: We examined the effects of FL on scopolamine (Sco)- and ethanol (EtOH)-induced hippocampus-dependent memory impairment, using the Passive avoidance (PA) and Morris water maze (MWM) tests. To examine the underlying mechanisms associated with neuroprotective effects, we analyzed acetylcholine (ACh) and acetylcholinesterase (AChE) activity, brain tissue expression of muscarinic acetylcholine receptor (mAChR), cAMP response element binding protein (CREB) and extracellular signal-regulated kinases 1/2 (ERK1/2), and immunohistochemical analysis, in the hippocampus of mice, compared to current drug therapy intervention. Biochemical blood analysis was carried out to determine the effects of FL on alanine transaminase (ALT), aspartate transaminase (AST), and triglyceride (TG) and total cholesterol (TC) levels. 7 groups (n = 10) consisted of a control (CON), 3 Sco-induced dementia and 3 EtOH-induced dementia groups, with both dementia group types containing an untreated group (Sco and EtOH); a positive control, orally administered donepezil (Dpz) (4mg/kg) (Sco + Dpz and EtOH + Dpz); and an FL (50 mg/kg) treatment group (Sco + FL50 and EtOH + FL50), orally administered over the 4-week experimental period. RESULTS: FL50 significantly reduced EtOH-induced increase in AST and ALT levels. FL50 treatment reduced EtOH-impaired step-through latency time in the PA test, and Sco- and EtOH-induced dementia escape latency times in the MWM test. Moreover, anticholinergic effects of Sco and EtOH on the brain were reversed by FL50, through the attenuation of AChE activity and elevation of ACh concentration. FL50 elevated ERK1/2 protein expression and increased p-CREB (ser133) in hippocampus brain tissue, according to Western blot and immunohistochemistry analysis, respectively. CONCLUSION: Overall, these results suggest that FL may be considered an efficacious intervention for Sco- and EtOH-induced dementia, in terms of reversing cognitive impairment and neuroplastic dysfunction.

Proteome Profiling of Murine Macrophages Treated with the Anthrax Lethal Toxin (탄저 치사독소 처리에 의한 생쥐 대식세포의 단백질체 발현 양상 분석)

  • Jung Kyoung-Hwa;Seo Giw-Moon;Kim Sung-Joo;Kim Ji-Chon;Oh Seon-Mi;Oh Kwang-Geun;Chai Young-Gyu
    • Korean Journal of Microbiology
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    • v.41 no.4
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    • pp.262-268
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    • 2005
  • Intoxication of murine macrophages (RAW 264.7) with the anthrax lethal toxin (LeTx 100 ng/ml) results in profound alterations in the host cell gene expression. The role of LeTx in mediating these effects is unknown, largely due to the difficulty in identifying and assigning function to individual proteins. In this study, we have used two-dimensional polyacrylamide gel electrophoresis to analyze the protein profile of murine macrophages treated with the LeTx, and have coupled this to protein identification using MALDI-TOF mass spectrometry. Interpretation of the peptide mass fingerprint data has relied primarily on the ProFound database. Among the differentially expressed spots, cleaved mitogen-activated protein kinase kinase (Mek1) and glucose-6-phosphate dehydrogenase were increased in the LeTx treated macrophages. Mek1 acts as a negative element in the signal transduction pathway, and G6PD plays the role for the protection of the cells from the hyper-production of active oxygen. Our results suggest that this proteomic approach is a useful tool to study protein expression in intoxicated macrophages and will contribute to the identification of a putative substrate for LeTx.

Identification and Molecular Characterization of Three Isoforms of Iturin Produced by Endophytic Bacillus sp. CY22 (식물 내생균 Bacillus sp. CY22가 생성하는 iturin isoform의 분리 및 특성)

  • Cho, Soo-Jeong;Yun-Han-Dae
    • Journal of Life Science
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    • v.15 no.6 s.73
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    • pp.1005-1012
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    • 2005
  • Endophytic Bacillus sp. CY22 was previously isolated from the interior of balloon flower root and showed strong antifungal activity against phytopathogenic fungi such as Rhizoctonia solnni, Fusarium oxysporum, and Phythium ultimum. Many Bacillus strains produce antifungal compound such as iturin, fengycin, and mycosubtilin. We isolated and identified antifungal compound from cell supernatant of the endophytic strain. By the MALDI-TOF mass result, the antifungal compound was similar to the known antifungal lipopeptide iturin. It was found that the purified iturin had three isoforms with protonated masses of m/z 1,043.39, 1,057.42, and 1,071.42 and different structures in combination with $Na^{+}$ ion using MALDI-TOF MS. The ita22 gene, which transacylase gene is associated with production of antifungal iturin, had an open reading frame (ORF) of 1,200 bp encoding 400 amino acids. Results of deduced amino acids sequence homology search, Ita22 was homologous with FenF (BAB69697) of Bacillus subtilis 168.

Cloning of α-Amylase Gene from Unculturable Bacterium Using Cow Rumen Metagenome (소 반추위 메타게놈에서 비배양 세균의 α-amylase 유전자 클로닝)

  • Cho, Soo-Jeong;Yun-Han-Dae
    • Journal of Life Science
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    • v.15 no.6 s.73
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    • pp.1013-1021
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    • 2005
  • The metagenomes of complex microbial communities are rich sources of novel biocatalysts. The gene encoding an extracellular $\alpha$-amylase from a genomic DNA of cow rumen was cloned in Escherichia coli DH5$\alpha$ and sequenced. The $\alpha$-amylase (amyA) gene was 1,893 bp in length, encoding a protein of 631 amino acid residues with calculated molecular weight of 70,734 Da. The molecular weight of the enzyme was estimated to be about 71,000 Da by active staining of a SDS-PACE. The enzyme was 21 to $59\%$ sequence identical with other amyloyltic enzymes. The AmyA was optimally active at pH 6.0 and $40\%$. The AmyA had a calculated pI of 5.87. AmyA expressed in E. coli DH5$\alpha$ was enhanced in the presence of $Mg^{2+}$ (20 mM) and $Ca^{2+}$ (30 mM) and inhibited in the presence of $Fe^{2+}$ and $Cu^{2+}$. The origin of amyA gene could not be confirmed by PCR using internal primer of amyA gene from extracted genomic DNA of 49 species rumen culturable bacteria so far. An amyh is supposed to obtained from unculturable rumen bacterium in cow rumen environment.

Status of Employment-Related Qualifications Similar to a Medical Laboratory Technology Major (임상병리기술학 전공 유사 취업연계 자격 현황)

  • Sung, Hyun Ho;Kim, Dae Sik;Cho, Young Kuk;Yoon, Ki Nam
    • Korean Journal of Clinical Laboratory Science
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    • v.50 no.4
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    • pp.525-534
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    • 2018
  • This study evaluated the various qualifications that can be linked to employment similar to the medical laboratory technology major. The qualifications for employment outside the medical laboratory technology major are industrial health instructor, doping control officer, audiologist, sign language interpreter, health education specialist, claim adjuster, Korea life underwriter, administrative manager, hospital administrator, insurance review manager, and hospital coordinator. The qualifications for employment similar to the medical laboratory technology major include the clinical research coordinator, clinical research associate, anatomist, analytical chemistry analysts, hazardous materials industry engineers, biotechnologist, biosafety managers, biotechnology technicians, medical device quality officers, animal care nurse, industrial engineer hazardous material, bioprocess engineer, biosafety officer, certified technology consultant, director of medical device quality control, laboratory animal technician, animal nursing technician, and cruise medical manager. Therefore, it is necessary to conduct a social analysis survey of the various qualifications currently held by medical technologists. In addition, it will be necessary to investigate the current status of medical technologists working in other fields. In the future, medical technologists should expand the scope of their work through efforts to strengthen their individual work capacity, share cases, and strengthen their expertise.