High-Level Production of Prourokinase-Annexin V Chimeras in the Methylotrophic Yeast Pichia pastoris (BIOCHEMICAL ENGINEERING)
スポンサーリンク
概要
- 論文の詳細を見る
Prourokinase (proUK)-annexin V chimeras expressed by the methylotrophic yeast Pichia pastoris in a synthetic medium as part of a system designed to yield a novel thrombolytic agent are degraded, as it is thought, by various yeast proteases present in the culture supernatant. Minimization of proteolysis was therefore investigated to increase the yield of intact proUK-annexin V. Protease inhibitor screening study indicated several proteases including at least serine protease like chymotrypsin were involved in the proteolysis. Addition of more than 10% of peptone or more than 0.2 mol l^-1 of arginine to the medium was effective in minimizing proteolysis in shake-flask culture. Culture condition of higher pH was also effective, however, induced a cell death. Cell improvement by increasing the methanol utilization ability yielded greater tolerance to high pH. As a result, the culture condition with highly concentrated peptone solution fed under controlled conditions of pH 8.0 was established, which greatly reduced proteolytic degradation in fed-batch fermentation. These optimal conditions, which enabled fibrinolytic activity to reach 7800 IU ml^-1,could easily be applied in industrial scale production.
- 社団法人日本生物工学会の論文
- 2002-11-25
著者
-
Ohya Tomoshi
Protein Research Laboratory, Mitsubishi Tanabe Pharma Corporation
-
Kobayashi Kaoru
Protein Research Laboratory, Mitsubishi Tanabe Pharma Corporation
-
Ohya Tomoshi
Protein Research Laboratory Pharmaceutical Research Division Mitsubishi Pharma Corporation
-
Ohya Tomoshi
大阪大学 工学研究科生命先端工学専攻
-
Ohya Tomoshi
Protein Reserach Laboratories Pharmaceuticals Research Division Mitsubishi Pharma Corporation
-
Morita M
Project Group Of Risk Assessment And Management For Endocrine Disruptor And Dioxin-like Compounds Na
-
Kuwae Shinobu
Protein Research Laboratory, Pharmaceutical Research Unit, Mitsubishi Pharma Corporation
-
MORITA MASANORI
Protein Reserach Laboratories, Pharmaceuticals Research Division, Mitsubishi Pharma Corporation
-
MIURA MASAMI
Protein Reserach Laboratories, Pharmaceuticals Research Division, Mitsubishi Pharma Corporation
-
Morita Masatomo
Department Of Bioengineering Tokyo Institute Of Technology
-
Ohda Toyoo
Bioscience Research Drug Discovery Laboratories Pharmaceutical Research Division Yoshitomi Pharmaceu
-
Kobayashi Kaoru
Protein Research Laboratory Pharmaceutical Research Division Mitsubishi Pharma Corporation
-
Kuwae Shinobu
Protein Research Laboratory Pharmaceutical Research Division Mitsubishi Pharma Corporation
-
Kuwae Shinobu
Research Center For Cell And Tissue Culture Faculty Of Agriculture Kyoto University
-
Morita M
Department Of Bioscience And Biotechnology Faculty Of Engineering Okayama University
-
Miura Masami
Protein Reserach Laboratories Pharmaceuticals Research Division Mitsubishi Pharma Corporation
-
Kobayashi Kaoru
Protein Research Laboratory Mitsubishi Tanabe Pharma Corporation
-
Ohya Tomoshi
Protein Research Laboratory Mitsubishi Tanabe Pharma Corporation
関連論文
- Overexpression of GADD34 Enhances Production of Recombinant Human Antithrombin III in Chinese Hamster Ovary Cells(CELL AND TISSUE ENGINEERING)
- Fate of Coliphage in a Wastewater Treatment Process
- Antibacterial Activity of Bacillus amyloliquefaciens Phage Endolysin without Holin Conjugation
- Identification of Genes from Aspergillus oryzae That Are Preferentially Expressed in Membrane-Surface Liquid Culture(GENETICS, MOLECULAR BIOLOGY, AND GENE ENGINEERING)
- Characteristics of α-Glucosidase Production from Recombinant Aspergillus oryzae by Membrane-Surface Liquid Culture in Comparison with Various Cultivation Methods(MICROBIAL PHYSIOLOGY AND BIOTECHNOLOGY)
- Development of a Fed-Batch Culture Process for Enhanced Production of Recombinant Human Antithrombin by Chinese Hamster Ovary Cells(CELL AND TISSUE ENGINEERING)
- Production of Recombinant Human Antithrombin by Pichia pastoris(Microbial Physiology and Biotechnology)
- High-Level Production of Prourokinase-Annexin V Chimeras in the Methylotrophic Yeast Pichia pastoris (BIOCHEMICAL ENGINEERING)
- High Level Secretion of Recombinant Human Serum Albumin by Fed-Batch Fermentation of the Methylotrophic Yeast, Pichia pastoris, Based on Optimal Methanol Feeding Strategy
- Addition of Oleic Acid Increases Expression of Recombinant Human Serum Albumin by the AOX2 Promoter in Pichia pastoris
- High-Level Expression of Recombinant Human Serum Albumin from the Methylotrophic Yeast Pichia pastoris with Minimal Protease Production and Activation
- Kinetic Analysis of Synthesis of a Dipeptide Precursor Using an Immobilized Enzyme in Water-Immiscible Organic Solvents
- Kinetics and Equilibrium for Thermolysin-Catalyzed Syntheses of Dipeptide Precursors in Aqueous/Organic Biphasic Systems
- Analysis for Partition Equilibrium of Amino Acid Derivatives in Aqueous/Organic Biphasic Systems
- THE EFFECTS OF CODING REGION ON ITS GENE EXPRESSION IN THE Chlamydomonas CHLOROPLAST
- Effect of Coding Regions on Chloroplast Gene Expression in Chlamydomonas reinhardtii
- Microbial Treatment of Bis (2-Ethylhexly) Phthalate in Polyvinyl Chloride with Isolated Bacteria (Environmental Biotechnology)
- Derivation of a Glycerol-negative Mutant of a Methylotrophic Yeast, Candida boidinii, and Accumulation of Glycerol in Methanol Medium(Microbiology & Fermentation Industry)
- Stability of Immobilized Thermolysin in Organic Solvents
- Overexpression of GADD34 Enhances Production of Recombinant Human Antithrombin III in Chinese Hamster Ovary Cells
- Production of Glycerol from Methanol by a Mutant Strain of Candida boidinii No. 2201