• Title/Summary/Keyword: microbial cell growth

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Bioconversion of Pinoresinol Diglucoside from Glucose Using Resting and Freeze-Dried Phomopsis sp. XP-8 Cells

  • Gao, Zhenhong;Rajoka, Muhammad Shahid Riaz;Zhu, Jing;Zhang, Zhiwei;Zhang, Yan;Che, Jinxin;Xu, Xiaoguang;Shi, Junling
    • Journal of Microbiology and Biotechnology
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    • v.27 no.8
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    • pp.1428-1440
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    • 2017
  • Phomopsis sp. XP-8 (an endophytic fungus) was previously found to produce pinoresinol diglucoside (PDG), a major antihypertensive compound of Tu-Chung (the bark of Eucommia ulmoides Oliv.), which is widely used in Chinese traditional medicines. In the present study, two bioconversion systems were developed for the production of PDG in Tris-HCl buffer containing glucose and Phomopsis sp. XP-8 cells (both resting and freeze-dried). When other factors remained unchanged, the bioconversion time, glucose concentration, cell ages, cell dosage, pH, temperature, and stirring speed influenced PDG production in a similar and decreasing manner after an initial increase with increasing levels for each factor. Considering the simultaneous change of various factors, the optimal conditions for PDG production were established as 70 g/l cells (8-day-old), 14 g/l glucose, $28^{\circ}C$, pH 7.5, and 180 rpm for systems employing resting cells, and 3.87 g/l cells, 14.67 g/l glucose, $28^{\circ}C$, pH 7.5, and 180 rpm for systems employing freeze-dried cells. The systems employing freeze-dried cells showed lower peak PDG production ($110.28{\mu}g/l$), but at a much shorter time (12.65 h) compared with resting cells (23.62 mg/l, 91.5 h). The specific PDG production levels were 1.92 and $24{\mu}g$ per gram cells per gram glucose for freeze-dried cells and resting cells, respectively. Both systems indicated a new and potentially efficient way to produce PDG independent of microbial cell growth.

Development of Transgenic Carrot Oral Vaccine to Protect against Diarrhea of Piglets (자돈 설사병 방지를 위한 경구백신용 형질전환 당근 개발)

  • 이영선;황철호
    • Journal of Plant Biotechnology
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    • v.29 no.4
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    • pp.287-293
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    • 2002
  • We are trying to develop a transgenic carrot with aims of production and delivery of oral vaccine against microbial enteropathogen using a K88ac pilin gene. A K88ac antigen (pilin) gene was isolated by PCR from the K88ac genomic DNA. The pilin gene was constructed in pGA748 and introduced via Agrobacterium tumefaciens to the explants of carrot hypocotyl and then 494 transgenic lines were established. The amounts of the K88ac antigen produced in each of the cell lines were determined by western and two elite cell lines (M1-17, Y14-1) were selected based on higher levels of expression of the antigens as well as rate of cell growth and efficiency of embryogenesis. In order to test an immunization induced by oral administration of the transgenic carrot, serum of the mice fed with the carrot vaccine were tested in ELISA. It tumed out that the mice fed with 3 g of transgenic carrot showed a similar level of antibody compared to those applied with 10 $\mu\textrm{g}$ of the purified recombinant pilin protein. Besides, various clinical responses were measured after challenging with ETEC K88ac strain to the piglets experiencing an oral immunization with the transgenic carrot. The piglets fed with carrot vaccine showed a lower level of diarrhea in fecal score compared to those fed with non-transgenic carrot. A higher level of increase in weight of the piglets fed with the transgenic carrot vaccine was observed comparing to those fed with non-transgenic carrot as control.

The Effects of Environmental Factors on Biodegradability Test for Lubricant Products (환경인자가 윤활제품의 생분해도 시험에 미치는 영향)

  • Cho, Eun-Hye;Park, Keun-Hyoung;Han, Seung-Ock;Kim, Eui-Yong;Ryu, Jae-Sang;Jang, Sun-Bok;Lee, Un-Gi;Chae, Hee-Jeong
    • KSBB Journal
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    • v.23 no.4
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    • pp.285-290
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    • 2008
  • Various methods (OECD 301B, ISO 9439 and ASTM 5864) for biodegradability test of lubricants were reviewed, and a standard procedure was developed. Most lubrication products are released in rivers or sea then is degraded by microbial action in aerobic condition. Most international method are based on $CO_2$ evolution test. Inoculum obtained from a sewage disposal plant and test compound are cultivated in a mineral medium. Organic carbon of the test compound is degraded and oxidized through the enzymatic actions of inoculum, and ultimately mineralized to carbon dioxide. Biodegradability test conditions of lubricant oils were optimized. The highest biodegradability was achieved when the same medium as in ASTM 5864 and inoculum concentration of $10^4{\sim}10^5$ cell/L were used. The optimum standard materials were selected as aniline and sodium acetate. Additionally the effects of inoculum type on microbial growth and biodegradability were examined. Finally the standard operating procedure (SOP) for biodegradability test method was proposed.

Antimicrobial Activities of Botanical Antimicrobial Agent-Grapefruit Seed Extract Mixture for the Preparation of Anitimicrobial Packaging Paper (항균포장지 제조용 식물성 자몽종자추출물제재의 항균특성)

  • Cho Sung-Hwan;Kim Chul-Hwan;Park Woo-Po
    • Food Science and Preservation
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    • v.11 no.3
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    • pp.411-416
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    • 2004
  • Botanical antimicrobial agent-grapefruit seed extract mixture(BAAG), which could be applied to the preparation of antimicrobial packaging paper, was investigated in order to prove the preservative function of fruits and vegetables. HAAG showed remarkable antimicrobial effects against Fusarium solani Botrytis cinerea, Pencillium crustosum, Erwinia carotovora, Phoma destructiva and Alternaria radicina causing the postharvest decay of fruits and vegetables. We have examined that HAAG could inhibit the growth of microorganims when treated with more than 500 $\mu$g/mL concentration. The activities of HAAG were stable in the wide spectrum of pH and temperature. Direct visualization of microbial cells by using scanning electron microscope showed the loss of microbial cell membrane function, which was destroyed by treating with the dilute solutions of HAAG. We could confirm that HAAG be an antimicrobial agent for the preparation of antimicrobial packaging paper.

Wastewater Treatment Characteristics by Pseudomonas sp. BLP2052 and Flavobacterium sp. BLP20515 Isolated from Sewage (선별된 Pseudomonas sp. BLP2052와 Flavobacterium sp. BLP20515의 폐하수 처리 특성)

  • 박철환;최광근;임지훈;이상훈;김상용;이진원
    • KSBB Journal
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    • v.14 no.2
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    • pp.153-159
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    • 1999
  • Fifteen microbes have been isolated from Jangja pond in Kuri-Si, Kyeonggi-Do. Among them, two strains showed excellent COD removal from wastewater, which were named Pseudomonas sp. BLP2052 and Flavobacterium sp. BLP20515, respectively. Optimal pH and temperature for the cell growth were 7.0 and $30^{\circ}C$ for both strains. Pseudomonas sp. BLP2052 and Flavobacterium sp. BLP20515 was applied to the reactor to treat wastewater and 66.0% and 65.7% COD (chemical oxygen demand) removal was achieved, respectively. Comparing these results to the case of applying mixed microbes present in Jangja pond, COD removal rate was 15% less. But when adding the selected microbes to the wastewater containing mixed microbes, COD removal rate increased by 5%. After 84 hour operation, we achieved 85.6% COD removal. When inhibitors were added less than 100 ppm, during the microbial wastewater treatment, Fe, Zn, Al, phenol and Cr influenced microbial activity more deterioratively in order. In the case of over 300 pm, Cr, Fe, Zn, Al and phenol showed severe deteriorative effect in order.

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Effect of Botanical Antimicrobial Agent-Citrus Products on the Quality Characteristics during Kimchi Fermentation (식물성 천연항균소재를 첨가한 김치의 숙성 중 품질변화)

  • Cho Sung-Hwan;Lee Seung-Gheol;Park Wan-Soo
    • Food Science and Preservation
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    • v.12 no.1
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    • pp.8-16
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    • 2005
  • To develop natural antimicrobial agents for extending the self-life of Kimchi, the effect of botanical antimicrobial agent-citrus products(BAAC) on microorganisms related to Kimchi spoilage was investigated. The inhibitory effect of BAAC on microorganisms related to Kimchi spoilage was increased according to the concentration of BAAC. Antimicrobial activities of BAAC against microoiganisms related to Kimchi spoilage were remarkably high. The effect of BAAC on the cellular membrane function of microorganisms showed the perturbation of cells in the presence of BAAC. Direct isualization of microbial cells by using both transmission md scanning electron microscope showed microbial cell membrane was destroyed by treating with BAAC. It could be confirmed that BAAC completely inhibit the growth of the test strains. The pH of BAAC-added Kimchi was a little higher than that of the control through the fermentation period. Titratable acidify, vitamin C and viable cells in BAAC-added Kimchi were changed more slowly than those in the control. Sensory evaluation did not show any significant difference between $0.01\%$ BAAC-added Kimchi and the control that showed the best palatabilities during fermentation.

Effect of Organic Acid Treatment on the Quality Attributes of Buckwheat Sprout during Storage (유기산 전처리에 따른 메밀 새싹의 저장중 품질변화)

  • Chang, Su-Kyung;Lee, Hyun-Hee;Hong, Seok-In;Han, Young-So
    • Korean Journal of Food Science and Technology
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    • v.42 no.2
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    • pp.190-197
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    • 2010
  • Changes in the quality of buckwheat sprout treated with organic acid solutions such as ascorbic, citric and acetic acid were examined during storage in order to judge the feasibility of acid dipping as a pretreatment for extending shelf-life. Quality attributes of buckwheat sprout stored at $5^{\circ}C$ were measured in terms of microbial cell count, moisture content, soluble solids content, chromaticity, and sensory evaluation over different storage times. For microbial growth inhibition, combinations of 0.05% acetic acid and 0.5% citric acid or 1% ascorbic acid and 0.5% citric acid were more effective than other treatments. Slight changes were observed in moisture content among the treatments, whereas soluble solids content of each treatment was increased during storage. In chromaticity, the Hunter's a value of buckwheat sprout treated with acetic acid alone or acetic acid combinations showed the largest increase. Dipping treatment with ascorbic acid and citric acid produced only slight changes in color of the sprout. In the sensory evaluation of discoloration (head, stem and root), wilting and overall quality, ascorbic acid and citric acid treatments gave the highest scores. These results suggest that dipping treatment of buckwheat sprout with citric acid can confer more positive effects on storage stability than others.

Inhibitory Effects of Stewartia koreana Extracts on Pseudomonas aeruginosa Biofilm Formation (노각나무 추출물이 Pseudomonas aeruginosa의 바이오필름 형성에 미치는 영향)

  • Sang Gyun Lee;Hye Soo Kim;Soo Jeong Cho
    • Journal of Life Science
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    • v.33 no.11
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    • pp.936-943
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    • 2023
  • This study was conducted to investigate the potential of Stewartia koreana as anti-microbial materials. The branches, stems and leaves of S. koreana were extracted into 70% ethanol and their antibacterial activity against P. aeruginosa was confirmed. The leaf, branch and stems extracts (1 mg/disc) showed the antibacterial activity against P. aeruginosa and leaf extracts showed higher antibacterial activities than those from branch extracts. The MIC against P. aeruginosa was 0.8 mg/ml and showed bacteriostatic action. The inhibitory effects of extract on biofilm formation and gene expression related to biofilm formation of P. aeruginosa was determined by biofilm biomass staining, SEM and qRT-PCR analysis. The biofilm biomass and cell growth of P. aeruginosa in the cultures treated with 0.2~2.0 mg/ml of S. koreana leaf extracts were significantly decreased in a concentration-dependent manner. We observed that the extract had an inhibitory effect on the formation of P. aeruginosa biofilms at concentrations of 0.8 mg/ml by SEM. qRT-PCR analysis showed that the lasI and rh1I gene expression associated to quorum sensing (QS) in the cultures treated with 0.2~2.0 mg/ml of S. koreana leaf extracts were suppressed in a concentration-dependent manner. Based on the above results, it can be concluded that S. koreana leaf extracts can be used as anti-microbial material derived from natural materials, as demonstrated by the antibacterial action and inhibition of biofilm formation of P. aeruginosa by QS inhibition.

The Interactive Effect of These Bacterial Substrates on the Growth of Streptococcus gordonii, Fusobacterium nucleatum and Porphyromonas gingivalis (Streptococcus gordonii, Fusobacterium nucleatum 및 Porphyromonas gingivalis의 상호작용이 성장에 미치는 영향)

  • Kim, A-Reum;Jeong, Moon-Jin;Ahn, Yong-Soon;Kim, Mi-Na;Kim, Sung-Im;Lim, Do-Seon
    • Journal of dental hygiene science
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    • v.15 no.2
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    • pp.209-219
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    • 2015
  • In order to explore an effect of interaction of Streptococcus gordonii, Fusobacterium nucleatum and Porphyromonas gingivalis that are bacteria relevant to periodontal disease on its growth, the bacteria were incubated in trypticase soy hemin menadione broth at $37^{\circ}C$ $CO_2$ incubator for 7 days through anaerobic jar by single and co-culture with heat treated dead bacteria under anaerobic gas pack. In order to confirm growth level, absorbance was measured and for confirming colony structure and form, it was observed with scanning electron microscope. In order to confirm an effect on pathogenicity of P. gingivalis, real time reverse transcriptase polymerase chain reaction was implemented for expression analysis for rgpA gene that produces HRgpA which is gingipain. As a result, the following conclusion was obtained. Colony formation of S. gordonii and P. gingivalis was increased by other dead bacteria and in case of F. nucleatum, its colony formation was showed an aspect of being increased by dead bacterium of P. gingivalis but decreased by dead bacterium of S. gordonii. Therefore, it is considered that the strains being used for this study would affect interactively through bacterial cell itself as well as their interaction factor at the time of colony formation.

The Experimental Study on Antioxidant, Anti-inflammatory, Antipruritic and Antibacterial Effects of the Banchong-san (BCS) (반총산의 항산화, 항염증, 항소양증, 항균효능에 관한 실험 연구)

  • Cho, Eun-Jin;Jo, Seong-Hui;Yang, Seung-Jeong
    • The Journal of Korean Obstetrics and Gynecology
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    • v.34 no.3
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    • pp.29-48
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    • 2021
  • Objectives: Banchong-san (BCS) is a herbal formula composed of 13 korean medicinal herbs and is traditionally used to treat inflammatory diseases and pain. The object of this study was to research the antioxidant, anti-inflammatory, antipruritic and antimicrobial effects of the BCS in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. Methods: In this experiment, effects of BCS on the following four were measured as follows: (1) Anti-oxidative effects were evaluated by 1,1-diphenyl-2-picryl-hydrazyl (DPPH) Radical scavenging activity, 2,2-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) Radical scavenging activity. (2) Anti-inflammatory effects were evaluated by the production amount of Reactive oxygen species (ROS), Nitric oxide (NO), Interleukin-1β (IL-1β), Interleukin 6 (IL-6), tumor necrosis factor-α (TNF-α), Prostaglandin E2 (PGE2), inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2)(the previous two are "mRNA"), extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), p38 mitogen-activated protein kinases (p38), inhibitor of nuclear factor kappa B (IκBα), nuclear factor kappa B (NF-κB) (the previous five are "Protein") in LPS-Stimulated RAW 264.7 cells. (3)Antipruritic effects were evaluated by the production amount of histamine, Leukotriene B4 (LTB4), LeukotrieneC4 (LTC4) Levels in phorbol 12-myristate 13-acetate(PMA)/ionomycin-stimulated MC/9 mast cell. (4) Anti-microbial effects were evaluated by the growth suppression of Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa and Aspergillus niger. Results: The following results were obtained through each measurement: (1) DPPH Radical Scavenging Activity, ABTS Radical Scavenging Activity evoked a significant concentration-dependent increase. (2) ROS, NO, IL-1β, IL-6, TNF-α, PGE2 production amount, iNOS, COX-2 mRNA expression were significantly reduced in the BCS extraction group compared with the control group and significantly decreased the amount of ERK, JNK, p38, NF-κB Protein expression. The amount of IκB-α Protein Expression have increased significantly. (3) The amounts of histamine, LTB4, LTC4 were significantly decreased. (4) The antibacterial efficacy, BCS inhibited the growth of Escherichia coli, Pseudomonas aeruginosa at concentrations of 5 ㎍/ml, but did not suppress the growth of staphylococcus aureus and aspergillus niger. Conclusions: The experimental results show that BCS has anti-oxidant, anti-inflammatory, antipruritic and antimicrobial properties.