• Title/Summary/Keyword: extracellular production

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Production of extracellular protein from Bacillus sp. WY-60 (Bacillus sp. WY-60에 의한 균체외 단백질의 분비조건)

  • Park, Shin;Kwon, Oh-Jin
    • Applied Biological Chemistry
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    • v.36 no.1
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    • pp.11-16
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    • 1993
  • A bacteria strain producing extracellular protein was isolated and identified from soil samples, and the optimum conditions of producing protein were investigated. Eleven strains of bacteria were isolated from soil samples. Among which WY-60 strain showed a very strong capability of producing protein and identified as a Bacillus sp. The optimum composition of nutrient medium for the production of the protein by WY-60 was fructose 4.0%, polypeptone 1.0%, $NH_4NO_3$ 0.1%, $K_2HPO_4$ 0.1%, $MgSO_4{\cdot}7H_2O$ 0.005%, $CaCO_3$ 1.0% and the optimum pH and temperature were 8.0 and $30^{\circ}C$, respectively. The penicillin G and lincomycin added to the above medium were effective for the protein production of the WY-60, but other antibiotics were non-effective. The maximum production of protein was obtained after 5 days culture.

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Stimulation of the Extracellular Matrix Production in Dermal Fibroblasts by Areca catechu Extract (진피섬유모세포에서 대복피추출물의 세포외기질 합성 촉진 효과)

  • Lee, Min-Ho;Kim, Hyung-Jin;Jung, Hyun-Ah;Lee, Young-Keun
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.14 no.4
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    • pp.1857-1862
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    • 2013
  • Dermal fibroblasts produce the many components of the extracellular matrix (ECM) that are needed to maintain connective tissue integrity and repair tissue injuries. This study investigated the effects of Areca catechu extract (ACE) on dermal fibroblast cell activation. Cultured human dermal fibroblasts were treated with ACE, and then ECM production was determined by ELISA, Western blot and RT-PCR. ACE significantly accelerated the production of type 1 collagen, fibronectin, and transforming growth factor (TGF)-${\beta}1$ by ELISA and type 1 collagen by Western blot assay. ACE also increased the gene expression of COL1A1, TGF-${\beta}1$, keratinocyte growth factor (KGF) and insulin growth factor (IGF)-1. These results suggest that ACE has the potential to stimulate ECM production and that it might be suitable for maintaining skin texture.

Screening of Microorganisms Secreted High Efficient Enzymes and Properties of Enzymatic Deinking for Old Newsprint(IV) -Culture conditions of fungal extracellular enzyme production for biological deinking system- (고효율 효소를 분비하는 균주의 선발 및 신문고지의 효소탈묵 특성(제4보) -고지탈묵용 Fungal Cellulase와 Xylanase의 생산-)

  • Park Seong-Cheol;Kang Jin-Ha;Lee Yang-Soo
    • Journal of Korea Technical Association of The Pulp and Paper Industry
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    • v.37 no.1 s.109
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    • pp.38-46
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    • 2005
  • This study was focused on the optimum culture condition in CMCase, FPase and xylanase activities of two fungal strains that secret extracellular enzymes for using enzymatic deinking agent to old newsprint. The results of this study were as follows. When Fusarium pallidoroseum was grown on the medium, containing of rice bran+xylan $2.0\%,\;peptone\;0.6\%,\;KH_2PO_4\;0.075\%\;and\;MnSO_4\;0.06\%\;with\;pH\;9.0,\;at\;29^{\circ}C$ for 6 days, the quantitative degree of extracellular enzyme production was the highest. Optimum culture condition for Aspergillus niger was pH 5.0, $27^{\circ}C$ incubating temperature and 7 days incubation period on liquid medium, containing of CMC+xylan $2.5\%,\;yeast\;extract\;0.4\%,\;K_3PO_4\;0.05\%\;and\;CaCl_2+FeSO_4\;0.08\%$. Aspergillus niger was fairly higher FPase and xylanase activities than Trichoderma reesei ATCC 28217.

A Productive Replication of Hyphantria cunea Nucleopolyhedrovirus in Lymantria dispar Cell Line

  • Demir, Ismail;Demirbag, Zihni
    • Journal of Microbiology and Biotechnology
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    • v.16 no.10
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    • pp.1485-1490
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    • 2006
  • In this study, comparative replicational properties of Hyphantria cunea nucleopolyhedrovirus (HycuNPV) in Lymantria dispar (IPLB-LdElta) and Spodoptera frugiperda (IPLB-Sf21) cell lines were investigated. Our microscopic observations showed that cytopathic effects (CPEs) in LdElta cells appeared 12 h later than those in Sf21 cells. Whereas polyhedral inclusion bodies (PIBs) formed at 48 h postinfection (p.i.) in LdElta cells, it formed at 36 h p.i. in Sf21 cells. Extracellular virus production determined according to the 50% tissue culture infective dose ($TCID_{50}$) method in LdElta cells started about 12 h later when compared with Sf21 cells. Titers of extracellular virus in LdElta and Sf21 cells were calculated as $1.77{\times}10^9$ plaque forming units (PFU)/ml and $5.6{\times}10^9PFU/ml$, respectively, at 72 h p.i. We also showed that viral DNA replication began at 12 h p.i. in both cell lines. Viral protein synthesis was determined by SDS-polyacrylamide gel electrophoresis (PAGE) and polyhedrin synthesis was observed at 12 h p.i. in both cell lines. The results indicate that while the synthesis of macromolecules is 12 h later and production of extracellular virus is almost 3-fold lower in LdElta cells compared with those in Sf21 cells, the LdElta cell line is still a productive cell line for infection of HycuNPV.

Andrographolide Promotes the Stemness of Epidermal Cells through the Extracellular Signal-regulated Kinase (ERK) Pathway (Andrographolide의 Extracellular Signal-regulated Kinase Pathway (ERK)를 통한 상피 세포 줄기세포능 향상)

  • You, Jiyoung;Roh, Kyung-Baeg;Shin, Seoungwoo;Park, Deokhoon;Jung, Eunsun
    • Korean Journal of Pharmacognosy
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    • v.50 no.1
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    • pp.18-24
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    • 2019
  • Andrographolide, the main compound of Andrographis paniculata (A. paniculata), shows various biological properties including anti-viral, anti-inflammatory, anti-diabetic, and hepatoprotective effects. Our previous study has shown that A. paniculata extract exerts antiaging effects by activation of stemness in epidermal stem cells (EpSCs). In this study, we investigated the effect of andrographolide as a main compound of A. paniculata on EpSCs and its mechnism of action using several in vitro assays. Andrographolide increased the proliferation of EpSCs and induced cell cycle progression. Additionally, andrographolide increased VEGF production and the expression of stem cell markers integrin ${\beta}1$ and p63. Furthermore, phosphorylation levels of extracellular signal-regulated kinase 1/2 (ERK1/2), S6 ribosomal protein (S6RP) and Akt were increased by andrographolide. Taken together, these results indicate that andrographolide-induced proliferation of EpSCs is mediated by the ERK1/2, Akt-dependent pathway with increased production of VEGF and upregulated stemness through integrin ${\beta}1$ and p63.

Production of Ethanol Directly from Potato Starch by Mixed Culture of Saccharomyces cerevisiae and Aspergillus niger Using Electrochemical Bioreactor

  • Jeon, Bo-Young;Kim, Dae-Hee;Na, Byung-Kwan;Ahn, Dae-Hee;Park, Doo-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.18 no.3
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    • pp.545-551
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    • 2008
  • When cultivated aerobically, Aspergillus niger hyphae produced extracellular glucoamylase, which catalyzes the saccharification of unliquified potato starch into glucose, but not when grown under anaerobic conditions. The $K_m\;and\;V_{max}$ of the extracellular glucoamylase were 652.3 mg/l of starch and 253.3 mg/l/min of glucose, respectively. In mixed culture of A. niger and Saccharomyces cerevisiae, oxygen had a negative influence on the alcohol fermentation of yeast, but activated fungal growth. Therefore, oxygen is a critical factor for ethanol production in the mixed culture, and its generation through electrolysis of water in an electrochemical bioreactor needs to be optimized for ethanol production from starch by coculture of fungal hyphae and yeast cells. By applying pulsed electric fields (PEF) into the electrochemical bioreactor, ethanol production from starch improved significantly: Ethanol produced from 50 g/l potato starch by a mixed culture of A. niger and S. cerevisiae was about 5 g/l in a conventional bioreactor, but was 9 g/l in 5 volts of PEF and about 19 g/l in 4 volts of PEF for 5 days.

Effect of nitric oxide on the expression of matrix metalloproteinases by the UV irradiated human dermal fibroblasts

  • Taeboo Choe;Lee, Bumchun;Park, Inchul;Seokil Hong
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.28 no.1
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    • pp.31-41
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    • 2002
  • The production of matrix matalloproteinases(MMPs) by the UV irradiated skin fibroblast and the degradation of extracellular matrix(ECM) by these enzymes is known as one of the main reasons of photoaging. Recently, Fisher group showed that the MMP expression is mainly regulated by the mitogen-activated protein(MAP) kinas family, such as extracellular signal-regulated kinase(ERK), c-Jun amino-terminal kinase(JNK) and p38, each of which forms a signaling pathway. In this work we first examined the effect of nitric oxide (NO) on the production of MMP-1 and MMP-2 by the human dermal fibroblasts (HDFs). NO is a multifunctional messenger molecule generated from L-arginine and involved in many kinds of signaling pathway. We found that the treatment of HDF with NO donor, sodium nitroprusside (SNP) enhanced the expression of MMPs with or without UV irradiation and the treatment with nitric oxide synthase (NOS) inhibitors resulted in the significant decrease of MMPs production. From these results, we concluded that the production of MMPs by the UV irradiated HDF is regulated through the signaling pathway involving NO and cyclic GMP.

Biotransformation of Protopanaxadiol-Type Ginsenosides in Korean Ginseng Extract into Food-Available Compound K by an Extracellular Enzyme from Aspergillus niger

  • Jeong, Eun-Bi;Kim, Se-A;Shin, Kyung-Chul;Oh, Deok-Kun
    • Journal of Microbiology and Biotechnology
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    • v.30 no.10
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    • pp.1559-1566
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    • 2020
  • Compound K (C-K) is one of the most pharmaceutically effective ginsenosides, but it is absent in natural ginseng. However, C-K can be obtained through the hydrolysis of protopanaxadiol-type ginsenosides (PPDGs) in natural ginseng. The aim of this study was to obtain the high concentration of food-available C-K using PPDGs in Korean ginseng extract by an extracellular enzyme from Aspergillus niger KACC 46495. A. niger was cultivated in the culture medium containing the inducer carboxymethyl cellulose (CMC) for 6 days. The extracellular enzyme extracted from A. niger was prepared from the culture broth by filtration, ammonium sulfate, and dialysis. The extracellular enzyme was used for C-K production using PPDGs. The glycoside-hydrolyzing pathways for converting PPDGs into C-K by the extracellular enzyme were Rb1 → Rd → F2 → C-K, Rb2 → Rd or compound O → F2 or compound Y → C-K, and Rc → Rd or compound Mc1 → F2 or compound Mc → C-K. The extracellular enzyme from A. niger at 8.0 mg/ml, which was obtained by the induction of CMC during the cultivation, converted 6.0 mg/ml (5.6 mM) PPDGs in Korean ginseng extract into 2.8 mg/ml (4.5 mM) food-available C-K in 9 h, with a productivity of 313 mg/l/h and a molar conversion of 80%. To the best of our knowledge, the productivity and concentration of C-K of the extracellular enzyme are the highest among those by crude enzymes from wild-type microorganisms.

Secretory production of prosubtilisin YaB by a six extracellular protease-deficient mutant of Bacillus subtilis

  • Byun, Dae-Seok;Chang, Young-Chae;Kang, Myung-Hwa
    • Journal of Life Science
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    • v.11 no.1
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    • pp.42-46
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    • 2001
  • Subtilisin YaB, produced by alkalophilic Bacillus strain YaB, is an extracellular alkaline serine protease having 55% homology to subtilisin BPN'. It is synthesized as a 378-amino acid preproenzyme and secreted into the culture medium as a 265-amino acid mature protease. To examine the role of pro-sequence for the secretion of subtilisin YaB, we have studied the expression, in Bacillus subtilis, of a mutant preprosubtilisin YaB in which active site Ser214 is substituted with Cys. The use of a six protease-deficient strain, WB600, was required for its efficient production. The prosubtilisin YaB, thus produced, was indeed secreted into the culture medium and was processed to its mature form upon treatment with exogenously added active subtilisin YaB. From these results, we have concluded that the processing of pro-sequence is not essential for the secretion of the enzyme.

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