Proteolytic Activity and Recombinant Protein Production in Virus-Infected Sf-9 Insect Cell Cultures Supplemented with Carboxyl and Cysteine Protease Inhibitors

スポンサーリンク

概要

• 論文の詳細を見る

• In insect cell-baculovirus expression systems for recombinant protein production, it is sometimes necessary to supplement cultures with protease inhibitors to protect recombinant proteins aginst proteolysis. To date, however, there is no information available concerning protease activities in inhibitor-supplemented cultures. The aim of the present study was to investigate intracellular and extracelular protease activities in cultures of virus-infected Sf-9 insect cells which were supplemented with inhibitors against carboxyl and cysteine proteases produced during culture. Prior to the supplementation culture, the cell foxicity of several protease inhibitors was determined. As a result, pepstatin A(carboxyl protease inhibitor)and E64, cystain, leupeptin, and antipain(cystein protease inhibitors)tested in this study showed no apparent negative effects on the growth and viability of noninfected Sf-9 insect cells at low concentrations. In addition, E64 and pepstain A could rapidly permeate virus-infected Sf-9 cells and inhibit the respective intracellular protease activities. A virus-infected culture with a multiplicity of infection of 1 was carried out with E64 and pepstain A which were added to the culture medium at 2d post-infection. As a result of inhibitor supplementation, the cellular activity for recombinant protein biosynthesis was reduced by 5-30%. However, a significant reduction in carboxyl and cysteine protease activities was observed not only in the medium but also intracellularly. This is the first study that directly demonstrates a reduction in extracellular and intracellular protease activities in protease inhibitor-supplemented cultures of virus-infected insect cells.

• 社団法人日本生物工学会の論文
• 2001-09-25

著者

• SATO Wataru

  Department of Gastroenterology, Akita University School of Medicine

• 後藤 健彦

  広島大学大学院工学研究科

• Gotoh Takeshi

  Department of Engineering in Applied Chemistry, Akita University

• Kikuchi Ken-Ichi

  Department of Engineering in Applied Chemistry, Akita University

• Gotoh T

  Department Of Materials-process Engineering And Applied Chemistry For Environments Akita University

• Gotoh Takeshi

  Department Of Materials Engineering And Applied Chemistry Akita University

• Gotoh Takeshi

  Department Of Materials And Applied Chemistry Mining College Akita University

• MIYAZAKI Yoshinori

  Department for Energy Conversion, Osaka National Research Institute

• Kikuchi Ken-ichi

  Department Of Materials-process Engineering And Applied Chemistry For Environments Akita University

• Kikuchi Ken-ichi

  Department Of Engineering In Applied Chemistry Akita University

• Kikuchi Ken-ichi

  Department Of Materials And Applied Chemistry Mining College Akita University

• Kikuchi K

  Akita Univ. Akita

• Gotoh Takeshi

  Department Of Engineering In Applied Chemistry Akita University

• BENTLEY WILLIAM

  Center for Agricultural Biotechnology and Department of Chemical Engineering, University of Maryland

• Bentley William

  Center For Agricultural Biotechnology And Department Of Chemical Engineering University Of Maryland

• Miyazaki Yoshinori

  Department Of Materials-process Engineering And Applied Chemistry For Environments Akita University

• Sato Wataru

  Department Of Environmental And Health Science Tohoku Pharmaceutical University

• Sato Wataru

  Department Of Materials-process Engineering & Applied Chemistry For Environments Akita Universit

• Miyazaki Yoshinori

  Department For Energy Conversion Osaka National Research Institute

• Gotoh Takeshi

  Department Of Engineering And Applied Chemistry Graduate School Of Engineering And Resource Science

• KIKUCHI KEN-ICHI

  Department of Chemical Engineering for Resources, Akita University

関連論文

• Active Human Renin Production Using a Baculovirus Expression Vector System: An Effective Method for Preventing Excessive Proteolytic Degradation of Recombinant Proteins
• Prorenin Processing Enzyme (PPE) Produced by Baculovirus-Infected Sf-9 Insect Cells: PPE Is the Cysteine Protease Encoded in the AcMNPV Gene
• Sex difference in the liver of hepatocyte-specific Pten-deficient mice : A model of nonalcoholic steatohepatitis
• 材料・界面
• 材料・界面
• 感温性多孔質高分子ゲルの転移温度制御と有機スラリー脱水への応用
• 複合化感温性多孔質ゲルを用いた有機スラリーの濃縮 (特集 注目されるゲルテクノロジーの応用)
• 感温性多孔質ゲルの生成および膨潤・収縮特性に及ぼす合成条件の影響
• 長鎖の疎水性側鎖をもつ感温性多孔質ゲルの構造制御と温度応答性
• 長鎖の疎水性側鎖の結晶化を利用した多孔性ポリマーの合成
• スポンジ状感温性ゲル--工業的分離媒体への応用を目指した簡便な合成法
• Preparation of Molecular Imprinted Thermosensitive Gel Adsorbents and Adsorption/Desorption Properties of Heavy Metal Ions by Temperature Swing
• The Application of Room Temperature Molten Salt with Low Viscosity to the Electrolyte for Dye-Sensitized Solar Cell
• Re-Infection Profile of Baculoviruses to Sf-9 Insect Cells that Have Already Been Infected : Virus Binding and Recombinant Protein Production
• Analysis of Inactivation of AcMNPV under Various Conditions by the ELVA Method
• Purification and Characterization of Aspartic Protease Derived from Sf9 Insect Cells
• High-Level Expression of Recombinant Active Human Renin in Sf-9 Cells: Rapid Purification and Characterization
• Epimorphin expression and stellate cell status in mouse liver injury
• Genipin prevents fulminant hepatic failure resulting in reduction of lethality through the suppression of TNF-α production
• Ultrastructural study of biopsy specimens of acute fatty liver of pregnancy
• SEPARATION OF AMINO ACIDS BY ELECTRODIALYSIS WITH IONEXCHANGE MEMBRANES
• Isolation of Human Renin Inhibitor from Soybean: Soyasaponin I Is the Novel Human Renin Inhibitor in Soybean
• The Occurrence of Renin Inhibitor in Rice: Isolation, Identification, and Structure-Function Relationship
• Hepatic gene expression in hepatocyte-specific Pten deficient mice showing steatohepatitis without ethanol challenge
• Enzyme Therapy in Gaucher Disease Type 2 : An Autopsy Case
• Identification and Expression of a Missense Mutation (Y446C) in the Acid Sphingomyelinase Gene from a Japanese Patient with Type A Niemann-Pick Disease
• IMMOBILIZATION OF MITOCHONDRIAL MEMBRANE IN SEPHAROSE 4B VIA PROTEOLIPOSOME RECONSTITUTION AND ITS D-β-HYDROXYBUTYPATE DEHYDROGENASE A CTIVITY
• Immobilization of γ-Glutamuyl Transpeptidase, as Membrane Enzyme, in Gel Beads via Liposome Entrapment
• Immobilization of Biological Membranes by Sonication and Freeze-Thawing
• Probabilistic Characterization for Baculovirus-Infected Insect Cells Destined to Synthesize Progeny Viruses and Recombinant Protein and to Die
• Significant Increase in Recombinant Protein Production of a Virus-Infected Sf-9 Insect Cell Culture of Low MOI under Low Dissolved Oxygen Conditions(BIOCHEMICAL ENGINEERING)
• Proteolytic Activity and Recombinant Protein Production in Virus-Infected Sf-9 Insect Cell Cultures Supplemented with Carboxyl and Cysteine Protease Inhibitors
• SORPTION OF AMINO ACIDS BY ION EXCHANGE MEMBRANES
• Active Human Renin Production Using a Baculovirus Expression Vector System : An Effective Method for Preventing Excessive Proteolytic Degradation of Recombinant Proteins
• 感温性高分子ゲルを用いた新規な物質分離法
• Purification and Properties of Extracellular Carboxyl Proteinase Secreted by Candida pulcherrima
• MEASUREMENT OF LOCAL RESIDENCE TIME OF PARTICLES IN A DRAFT-TUBE SLURRY BUBBLE COLUMN USING MAGNETIC TRA CER PARTICLES
• Isolation of Human Renin Inhibitor from Soybean : Soyasaponin I Is the Novel Human Renin Inhibitor in Soybean
• Purification and Characterization of Aspartic Protease Derived from Sf9 Insect Cells
• Construction of a System that Simultaneously Evaluates CYP1A1 and CYP1A2 Induction in a Stable Human-derived Cell Line using a Dual Reporter Plasmid
• The Occurrence of Renin Inhibitor in Rice : Isolation, Identification, and Structure-Function Relationship
• High-Level Expression of Recombinant Active Human Renin in Sf-9 Cells : Rapid Purification and Characterization
• A New Access to Mori's Intermediate for the Practical Synthesis of (-)-Periplanone-B
• A Controlled Synthesis of 9α-Methyl- and 9α-Cyanoisocarbacyclins
• Prorenin Processing Enzyme (PPE) Produced by Baculovirus-Infected Sf-9 Insect Cells : PPE Is the Cysteine Protease Encoded in the AcMNPV Gene
• Analysis of Inactivation of AcMNPV under Various Conditions by the ELVA Method
• Room Temperature Molten Salts Based on Tetraalkykammonium Cations and Bis(trifluoromethylsulfonyl)imide
• Recovery of Rare-metal Elements from their EDTA Complex Solution by means of Electrodialysis Accompanied by Metal Substitution Reaction (「日中電気透析シンポジウム」小特集)
• Effect of Clearance between Two Sensor-Probes on Measured Values of Bubble Properties in a Bubble Column
• Room Temperature Molten Salts Based on Trialkylsulfonium Cations and Bis(trifluoromethylsulfonyl)imide
• Chitinase from Autographa californica Multiple Nucleopolyhedrovirus: Rapid Purification from Sf-9 Medium and Mode of Action
• S-3 Practical use of mass media : Merit and demerit of the health Information TV programs(The Proceedings of the 15th Annual Meetings of Japan Society of Exercise and Sports Physiology July 26-27, (Hirosaki))
• Chitinase from Autographa californica Multiple Nucleopolyhedrovirus : Rapid Purification from Sf-9 Medium and Mode of Action
• Osler-Weber-Rendu Disease with Esophageal Varices and Hepatic Nodular Change
• Efficacy of Contrast-Enhanced Ultrasonography in Radiofrequency Ablation for Hepatocellular Carcinoma
• Mass transfer between particles and liquid in solid-liquid two-phase upflow in vertical tubes.
• Average particle velocity in solid-liquid two-phase flow through vertical and horizontal tubes.
• Correlation of mass transfer coefficient between particles and liquid in liquid fluidized beds.
• Mass transfer between particles and liquid in dilute fluidized beds in the low Reynolds number region.
• Correlation of liquid-side mass transfer coefficient for single particles and fixed beds.
• Axial dispersion of liquid in liquid fluidized beds in the low Reynolds number region.
• Mass transfer between particles and liquid in solid-liquid two-phase upflow in the low-velocity region through vertical tubes.
• Mass transfer between particles and liquid in solid-liquid two-phase flow through horizontal tubes.
• Isolation and Identification of a New Type of β-Conglycinin in Soybean Globulins

もっと見る 閉じる

スポンサーリンク