• Title/Summary/Keyword: extracellular protein

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Seasonal Fluctuations of Heterotrophic Activity and Bacterial Extracellular Enzyme Activity in Paldang Lake (팔당호에서 종속영양 활성도의 계절적 변화 및 세균의 세포외 효소활성)

  • 김상진
    • Korean Journal of Microbiology
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    • v.31 no.1
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    • pp.93-98
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    • 1993
  • To investigate the organic matter transformation in aquatic environment, seasonal fluctuations of heterotrophic activity and microbia] extracellular enzyme activity were studied in Paldang Lake, Korea. The turnover time in the water column and the sediment at the station I fluctuated between 3 -I ,300 hrs and 17-170 hrs for glucose, 5 -1.900 hrs and 15-240 hrs for protein hydrolysate and 4-350 hrs and 15-230 hrs for acetic acid, respectively, indicating that the seasonal turnover time of organic substrates fluctuated drastically. The respiration ratios of glucose. protein hydrolysate and acetate were 23-32%, 38-41% and 22-28% in the water column and 34%, 61% and 41% in the sediment. respectively. These results showed that the respiration ratios in the sediment were higher than those in the water column regardless of kinds of organic substrates. The bacterial extracellular enzyme activities of $\alpha$-glucosidase. $\beta$-glucosidase, N-acetyl-$\beta$-D-glucosaminidase and aminopeptidase were 32-44%. 31-32%, 18-34% and 61-67% in the water column, and 34%. 40%, 23% and 65% in the sediment. respectively.

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Optimization of Extracellular Production of Recombinant Human Bone Morphogenetic Protein-7 (rhBMP-7) with Bacillus subtilis

  • Kim, Chun-Kwang;Rhee, Jong Il
    • Journal of Microbiology and Biotechnology
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    • v.24 no.2
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    • pp.188-196
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    • 2014
  • Extracellular production of recombinant human bone morphogenetic protein-7 (rhBMP-7) was carried out through the fermentation of Bacillus subtilis. Three significant fermentation conditions and medium components were selected and optimized to enhance the rhBMP-7 production by using the response surface methodology (RSM). The optimum values of the three variables for the maximum extracellular production of rhBMP-7 were found to be 2.93 g/l starch, 5.18 g/l lactose, and a fermentation time of 34.57 h. The statistical optimization model was validated with a few fermentations of B. subtilis in shake flasks under optimized and unoptimized conditions. A 3-L jar fermenter using the shake-flask optimized conditions resulted in a higher production (413 pg/ml of culture medium) of rhBMP-7 than in a shake flask (289.1 pg/ml), which could be attributed to the pH being controlled at 6.0 and constant agitation of 400 rpm with aeration of 1 vvm.

The Extracellular Enzyme Activities in Culture Broth of Sparassis crispa. (꽃송이버섯(Sparassis crispa)의 세포외 효소활성)

  • Kim Ji-Young;Lim Chang-Soo;Kim Jae-Yong;Han Yeong-Hwan
    • Korean Journal of Microbiology
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    • v.40 no.3
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    • pp.230-231
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    • 2004
  • The mycelia of Sparassis crispa DSMZ 5201 were cultivated at $24^{\circ}C$ for 15 days in yeast-malt extract-glucose broth (pH 4.0) and the filtrate was used as crude enzyme solution to determined the extracellular enzyme activity. The specific activity of $\alpha$-amylase was 44.27 unit/protein. The specific activities of protease, CMCase, $\beta$-glucosidase, chitinase, exo-$\beta$-l,4-glucanase were relatively high. However, a very little activity of xylanase was found.

Expression and Purification of Recombinant Human Bone Morphogenetic Protein-7 (rhBMP-7) in Bacillus subtilis (고초균을 이용한 재조합 인간 골 형성 단백질-7의 발현과 정제)

  • Kim, Chun-Kwang;Oh, Sung-Duk;Rhee, Jong-Il
    • KSBB Journal
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    • v.25 no.3
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    • pp.257-264
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    • 2010
  • Bone morphogenetic protein-7 (BMP-7) is one of important growth factors for skeletal development and bone growth. In this work, BMP-7 was efficiently expressed in recombinant Bacillus subtilis. The mature BMP-7 protein indicated molecular weight of 15.4 kDa by Western blot assay and was secreted into culture medium with 0.35 ng/mL. The extracellular and intracellular rhBMP-7 proteins were purified by using a FPLC system with an ion exchange column and a gel filtration column. The extracellular and intracellular rhBMP-7 proteins had finally a 57.1% purity and a 36.2% purity, respectively. The purified rhBMP-7 proteins showed an intact biological activity which stimulated alkaline phophatase (ALP) activity in MC3T3-E1 cells.

Transforming Stimulated Clone 22 (TSC-22) Interacts Directly with Bromodomain-Containing Protein 7 (BRD7) to Enhance the Inhibition of Extracellular Signal-Regulate Kinase (ERK) Pathway in Ovarian Cancer

  • Lee, Seung-Hoon;Choi, Donchan
    • Development and Reproduction
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    • v.26 no.3
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    • pp.117-126
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    • 2022
  • Bromodomain-containing protein 7 (BRD7) participates in many cellular processes and embryo development. BRD7 is down-regulated in various cancers and evidence of its tumor suppressor function has been accumulating. Here, we identified transforming stimulated clone 22 (TSC-22) as a novel BRD7 interacting protein and show its novel function as a positive regulator of BRD7. We found that TSC-22 expression potentiated the inactivation of the extracellular signal-regulate kinase (ERK) pathway by BRD7. Our data establishes TSC-22 as a modulator of BRD7 and unravels the molecular mechanisms that drive the synergistic tumor-suppressing effects of TSC-22 and BRD7. Our findings may open new avenues for developing novel molecular therapies for tumors exhibiting down-regulated BRD7 and/or TSC-22.

ANKS1A-Deficiency Aberrantly Increases the Entry of the Protein Transport Machinery into the Ependymal Cilia

  • Haeryung Lee;Jiyeon Lee;Miram Shin;Soochul Park
    • Molecules and Cells
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    • v.46 no.12
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    • pp.757-763
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    • 2023
  • In this study, we examine whether a change in the protein levels for FOP in Ankyrin repeat and SAM domain-containing protein 1A (ANKS1A)-deficient ependymal cells affects the intraflagellar transport (IFT) protein transport system in the multicilia. Three distinct abnormalities are observed in the multicilia of ANKS1A-deficient ependymal cells. First, there were a greater number of IFT88-positive trains along the cilia from ANKS1A deficiency. The results are similar to each isolated cilium as well. Second, each isolated cilium contains a significant increase in the number of extracellular vesicles (ECVs) due to the lack of ANKS1A. Third, Van Gogh-like 2 (Vangl2), a ciliary membrane protein, is abundantly detected along the cilia and in the ECVs attached to them for ANKS1A-deficient cells. We also use primary ependymal culture systems to obtain the ECVs released from the multicilia. Consequently, we find that ECVs from ANKS1A-deficient cells contain more IFT machinery and Vangl2. These results indicate that ANKS1A deficiency increases the entry of the protein transport machinery into the multicilia and as a result of these abnormal protein transports, excessive ECVs form along the cilia. We conclude that ependymal cells make use of the ECV-based disposal system in order to eliminate excessively transported proteins from basal bodies.

Thin Layer Chromatogram by an Extracellular ${\beta}$-Amylase of Bacillus sp. KYJ 963 and its Amino Acid Composition

  • Kim, Young-Jae
    • Journal of Life Science
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    • v.11 no.2
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    • pp.92-93
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    • 2001
  • Bacillus sp. KYJ 963, which was isolated from Korean salt-fermented anchovy (anchovy-jeot), produces an extracellular ${\beta}$-amylase. The analysis of the digestion products of substrates by thin layer chromatography from the purified protein revealed that the enzyme could not hydrolyze maltose or ${\alpha}$-cyclodextrin. In the amino acid composition analysis, the major characteristic of the ${\beta}$-amylase was the high proportion of amino acids that possess short side chain such as glycine and alanine.

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Characterization of α-D-manosidase activity from Bacillus safensis MA-01 (Bacillus safensis MA-01 유래 알파-만노사이데이즈의 효소학적 특성)

  • Lee, Bo Mi;Kim, Joo Won;Park, Jae Kweon
    • Journal of Marine Bioscience and Biotechnology
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    • v.7 no.1
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    • pp.11-18
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    • 2015
  • An extracellular alkaline ${\alpha}$-D-mannosidase produced by a strain named as MA-01 was produced and its preliminary enzyme activity was characterized. Upon determining the 16S rDNA sequence and its homology search, the strain was identified to be one of species of the Bacillus safensis. Localization of enzyme was elucidated that ${\alpha}$-D-mannosidase can be found in culture medium as an extracellular enzyme. In addition, partial enzyme activity of 63% compared with the extracellular enzyme activity was observed in membrane protein. The optimal pH and temperature of the ${\alpha}$-D-mannosidase were pH 7.5 and $37^{\circ}C$, respectively. The $K_m$ and $V_{max}$ values of the ${\alpha}$-D-mannosidase in crude enzyme toward p-nitrophenyl-${\alpha}$-D-mannopyranoside were determined to be $455.6{\mu}M$ and $10.8{\mu}mole/min/mg$ of protein, respectively. To the best of our knowledge, this is the first report described the alkaline ${\alpha}$-D-mannosidase from the family of B. safensis.

Extracellular synthesis of silver nanoparticle by Pseudomonas hibiscicola - Mechanistic approach

  • Punjabi, Kapil;Mehta, Shraddha;Yedurkar, Snehal;Jain, Rajesh;Mukherjee, Sandeepan;Kale, Avinash;Deshpande, Sunita
    • Advances in nano research
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    • v.6 no.1
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    • pp.81-92
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    • 2018
  • Biosynthesis of nanoparticles has acquired particular attention due to its economic feasibility, low toxicity and simplicity of the process. Extracellular synthesis of nanoparticles by bacteria and fungi has been stated to be brought about by enzymes and other reducing agents that may be secreted in the culture medium. The present study was carried out to determine the underlying mechanisms of extracellular silver nanoparticle synthesis by Pseudomonas hibiscicola isolated from the effluent of an electroplating industry in Mumbai. Synthesized nanoparticles were characterized by spectroscopy and electron microscopic techniques. Protein profiling studies were done using Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis (1D-SDS PAGE) and subjected to identification by Mass Spectrometry. Characterization studies revealed synthesis of 50 nm nanoparticles of well-defined morphology. Total protein content and SDS PAGE analysis revealed a reduction of total protein content in test (nanoparticles solution) samples when compared to controls (broth supernatant). 45.45% of the proteins involved in the process of nanoparticle synthesis were identified to be oxidoreductases and are thought to be involved in either reduction of metal ions or capping of synthesized nanoparticles.

Anti-wrinkle effect of bone morphogenetic protein receptor 1a-extracellular domain (BMPR1a-ECD)

  • Yoon, Byung-Hak;Jeon, Yun-Hui;Hwang, Byunghee;Kwon, Hyuknam;Choe, Senyon;Yang, Zungyoon
    • BMB Reports
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    • v.46 no.9
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    • pp.465-470
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    • 2013
  • Bone morphogenetic proteins (BMPs) have diverse and important roles in the proliferation and differentiation of adult stem cells in our tissues. Especially, BMPs are well known to be the main inducers of bone formation, by facilitating both proliferation and differentiation of bone stem cells. Interestingly, in skin stem cells, BMPs repress their proliferation but are indispensable for the proper differentiation into several lineages of skin cells. Here, we tested whether BMP antagonists have an effect on the prevention of wrinkle formation. For this study we used an in vivo wrinkle-induced mouse model. As a positive control, retinoic acid, one of the top anti-wrinkle effectors, showed a 44% improvement compared to the non-treated control. Surprisingly, bone morphogenetic protein receptor 1a extracellular domain (BMPR1a-ECD) exhibited an anti-wrinkle effect which was 6-fold greater than that of retinoic acid. Our results indicate that BMP antagonists will be good targets for skin or hair diseases.