• Title/Summary/Keyword: Extracellular

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Fungal Metabolism of Environmentally Persistent Compounds: Substrate Recognition and Metabolic Response

  • Wariishi, Hiroyuki
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.5 no.6
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    • pp.422-430
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    • 2000
  • Mechanism of lignin biodegradation caused by basidiomycetes and the history of lignin biodegradation studies were briefly reviewed. The important roles of fungal extracellular ligninolytic enzymes such as lignin and manganese peroxidases (LiP and MnP) were also summarized. These enzymes were unique in their catalytic mechanisms and substrate specificities. Either LiP or MnP system is capable of oxidizing a variety of aromatic substrates via a one-electron oxidation. Extracellular fungal system for aromatic degradation is non-specific, which recently attracts many people working a bioremediation field. On the other hand, an intracellular degradation system for aromatic compounds is rather specific in the fungal cell. Structurally similar compounds were prepared and metabolized, indicating that an intracellular degradation strategy consisted of the cellular systems for substrate recognition and metabolic response. It was assumed that lignin-degrading fungi might be needed to develop multiple metabolic pathways for a variety of aromatic compounds caused by the action of non-specific ligninolytic enzymes on lignin. Our recent results on chemical stress responsible factors analyzed using mRNA differential display techniques were also mentioned.

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Heterologous Gene Expression and Secretion of the Anticoagulant Hirudin in a Methylotrophic Yeast Hansenula polymorpha

  • Sohn, Jung-Hoon;Michael-Yu-Beburov;Choi, Eui-Sung
    • Journal of Microbiology and Biotechnology
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    • v.3 no.2
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    • pp.65-72
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    • 1993
  • A heterologous gene expression and secretion system using a methylotrophic yeast, Hansenula polymorpha was developed for the production of anticoagulant hirudin. Hirudin gene was expressed under the control of a strong and inducible methanol oxidase (MOX or AOX) promoter. The mating factor a pre-pro leader sequence of Saccharomyces cerevisiae was employed for hirudin to be secreted into the extracellular medium. Hirudin expression cassette was introduced into three strains of H. polymorpha, A16, HPBl and DLl which have different genetic backgrounds. This expression cassette was stably integrated into the host chromosomal DNA. Biologically active and mature hirudin was efficiently expressed and secreted into the extracellular medium. About 19 mg/L of hirudin was found in the culture supernatant in the case of a two-copy integrant of the strain HPBl under suboptimal culture conditions.

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Fluorescent and bioluminescent nanoprobes for in vitro and in vivo detection of matrix metalloproteinase activity

  • Lee, Hawon;Kim, Young-Pil
    • BMB Reports
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    • v.48 no.6
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    • pp.313-318
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    • 2015
  • Matrix metalloproteinases (MMPs) are zinc-dependent endopeptidases that degrade the extracellular matrix (ECM) and regulate the extracellular microenvironment. Despite the significant role that MMP activity plays in cell-cell and cell-ECM interactions, migration, and differentiation, analyses of MMPs in vitro and in vivo have relied upon their abundance using conventional immunoassays, rather than their enzymatic activities. To resolve this issue, diverse nanoprobes have emerged and proven useful as effective activity-based detection tools. Here, we review the recent advances in luminescent nanoprobes and their applications in in vitro diagnosis and in vivo imaging of MMP activity. Nanoprobes with the purpose of sensing MMP activity consist of recognition and detection units, which include MMP-specific substrates and luminescent (fluorescent or bioluminescent) nanoparticles, respectively. With further research into improvement of the optical performance, it is anticipated that luminescent nanoprobes will have great potential for the study of the functional roles of proteases in cancer biology and nanomedicine. [BMB Reports 2015; 48(6): 313-318]

Autophagy in neutrophils

  • Shrestha, Sanjeeb;Lee, Jae Man;Hong, Chang-Won
    • The Korean Journal of Physiology and Pharmacology
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    • v.24 no.1
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    • pp.1-10
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    • 2020
  • Autophagy is a highly conserved intracellular degradation and energy-recycling mechanism that contributes to the maintenance of cellular homeostasis. Extensive researches over the past decades have defined the role of autophagy innate immune cells. In this review, we describe the current state of knowledge regarding the role of autophagy in neutrophil biology and a picture of molecular mechanism underlying autophagy in neutrophils. Neutrophils are professional phagocytes that comprise the first line of defense against pathogen. Autophagy machineries are highly conserved in neutrophils. Autophagy is not only involved in generalized function of neutrophils such as differentiation in bone marrow but also plays crucial role effector functions of neutrophils such as granule formation, degranulation, neutrophil extracellular traps release, cytokine production, bactericidal activity and controlling inflammation. This review outlines the current understanding of autophagy in neutrophils and provides insight towards identification of novel therapeutics targeting autophagy in neutrophils.

Production of the Extracellular Alkaline Proteinase by Yarrowia Lipolytica 504D (Yarrowia lipolytica 504D의 Extracellular Alkaline Proteinase 생산성)

  • 유춘발;김창화;김태곤
    • Journal of Life Science
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    • v.8 no.3
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    • pp.333-338
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    • 1998
  • Productivity of alkaline proteinase from Yarrowia lipolytica 504D was investigated. For the production fo the enzyme, hemoglobin was the best nitogen source, however, casein and skim milk were also good. All carbon sources inhibited strongly the producitivity of the enzyme. Yeast extract increased the productivity of the enzyme to 220%, but almost mineral salts except monovalant ions decreased it. Based on these results, optimal medium was composed of 1.2% casein, 0.2% glucose, 0.16% yeast extract, and 0.1% ammonium sulfate. the best condition for the production of the enzyme was observed at pH 9 and $20^{\circ}C$ for 42 hours.

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Study on recombinant expression of Phospholipase C gene (plc) in methylotrophic yeast Pichia pastoris and its properties

  • Seo, Kook-Hwa;Rhee, Jong-Il
    • 한국생물공학회:학술대회논문집
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    • 2003.10a
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    • pp.191-194
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    • 2003
  • The phospholipase C (PLC) hydrolyzes the polar head groups such as phosphocholine or phosphoethanolamine residues esterified at the sn-3 position of phospholipids. Pichia pastoris can utilize methanol as a carbon source and produce recombinant proteins under the control of the strong, tightly-regulated alcohol oxidase (AOX) promoter. In this study, we developed recombinant P. pastoris system for PLC expression and analyzed PLC activity.

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Light Mediated Yeast Cell Growth and Metabolism

  • Dowd Jr, Christopher J.;Tanner, Robert D.
    • KSBB Journal
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    • v.6 no.3
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    • pp.299-307
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    • 1991
  • In this paper the effect of light on non-aerated Baker's Yeast(Saccharomyces cereuisiae) production and the protein excretion to the extracellular fluid is studied. Previous results in our laboratory indicate that at pH=5 and T-32$^{\circ}C$ yeast may be affected by light, but those differences seem to be within statistical variation of the data. In this paper, cell and extracellular protein concentrations along with redox potential are monitored for batch fermentations in the presence and absence of light at pH levels of 3 and 5 and at 31$^{\circ}C$, in order to explore whether possible light effects can be more readily discerned at lower pH values. Yeast particle size distributions are also determined over the course of fermentation using a particle counter in order to add one more measuring tool to our usual cell and total protein measurements. An apparently noticeable difference in the redox potential is observed between the light and the dark runs for early times for the pH=3 runs. The particle size distributions show differences in the particle diameters between light and dark runs at pH=3, but those differences fall within one standard deviation of the mean particle diameters.

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Characteristics of a Marine Agarolytic Pseudomonas sp. from Porphyra dentata(Bangiales, Rhodophyta) and Some Properties of its Extracellular Agarase (김(Porphyra dentata) 병반조직에서 분리한 해양미생물의 특성과 생산된 체외 한천분해효소 특성)

  • 박상렬;조수정;김민근;임우진;류성기;안창룡;홍수영;이영한;김범규
    • Journal of Life Science
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    • v.11 no.4
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    • pp.291-297
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    • 2001
  • The marine bacterium isolated from Porphyra dentata showing green spot rot disease was identified as Pseudomonas sp. the strain have CNCase activity, xylanase activity and protease activity as well as agarase activity. But the strain has no pectate lyase activity. Porphyra dentata tissue inoculated this isolate was macerated after 1 week incubation. The characteristics of extracellular crude agarase of this isolate were examined, the optimal pH and temperature were pH7 and 3$0^{\circ}C$, respectively.

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Methylotrophic yeast Pichia pastoris를 이용한 재조합 phospholipase C (PLC) 생산 및 특성 연구

  • Seo, Guk-Hwa;Jeong, Sang-Yun;Lee, Jong-Il;Bornscheuer, Uwe T.
    • 한국생물공학회:학술대회논문집
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    • 2003.04a
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    • pp.233-235
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    • 2003
  • The phospholipase C (PLC) hydrolyzes the polar head groups such as phosphocholine or phosphoethanolamine residues esterified at the sn-3 position of phospholipids. Pichia pastoris can utilize methanol as a carbon source and produce recombinant proteins under the control of the strong, tightly-regulated alcohol oxidase (AOX) promoter. In this study, we developed recombinant P. pastoris system for the high productivity of PLC and analyzed PLC activity.

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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.