当前位置： SCI文献检索 > PROTEIN EXPRESSION AND PURIFICATION期刊下所有文献 > Optimization of human D-amino acid oxidase expression in Escherichia coli.

Optimization of human D-amino acid oxidase expression in Escherichia coli.

Abstract：

:Human D-amino acid oxidase (hDAAO) is a flavoprotein that plays a key role in the pathophysiology of schizophrenia. So far, the biochemical characterization of this enzyme has been hampered by the difficulty of expressing it in a common heterologous host such as Escherichia coli. Increasing amounts of recombinant hDAAO are indeed required for the investigation of its structure-function relationships and for the screening of new inhibitors to be used in the treatment of schizophrenia. A recombinant hDAAO has been over-expressed in BL21(DE3)Star E. coli cells. By alternating screenings of medium components at flask level and investigating physiological parameters in 2L controlled batch fermentations, an improved, robust and scalable microbial process was set up giving almost a 40- and 4-fold improvement in volumetric productivity and specific activity, respectively. Under these conditions approximately 770 U/L culture hDAAO with a specific activity of approximately 0.4 U/mg protein and a specific productivity of 24.9 U/g biomass were produced. Optimization of medium ingredients, of the time and the amount of inducer's addition, pH control at the moment of induction and harvest, low mechanical shear stress regime during recombinant protein production, represent the factors concurring to achieve the reported expression level. Notably, this expression level is higher than any previously described production of hDAAOs. A yield of 100 mg of pure hDAAO/L culture thus became available in comparison to the 1-10 mg/L previously reported.

journal_name

Protein Expr Purif

journal_title

Protein expression and purification

authors

Romano D,Molla G,Pollegioni L,Marinelli F

doi

10.1016/j.pep.2009.05.013

subject

Has Abstract

pub_date

2009-11-01 00:00:00

pages

72-8

issue

1

eissn

1046-5928

issn

1096-0279

pii

S1046-5928(09)00136-3

journal_volume

68

pub_type

杂志文章

相关文献

PROTEIN EXPRESSION AND PURIFICATION文献大全
  • Cloning, expression, isotope labeling, and purification of human antimicrobial peptide LL-37 in Escherichia coli for NMR studies.

    abstract：:Antimicrobial peptide LL-37 plays an important role in human body's first line of defense against infection. To better understand the mechanism of action, it is critical to elucidate the three-dimensional structure of LL-37 in complex with bacterial membranes. We present a bacterial expression system that allows the i...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2005.10.022

    authors： Li Y,Li X,Wang G

    更新日期：2006-06-01 00:00:00

  • Expression and purification of functional, recombinant Trypanosoma cruzi complement regulatory protein.

    abstract：:The complement regulatory protein (CRP) of Trypanosoma cruzi is a developmentally regulated glycosylphosphatidylinositol (GPI)-anchored membrane protein that protects the parasite from complement-mediated killing, and is an important virulence determinant of the microorganism. CRP binds human complement components C3b...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/s1046-5928(02)00562-4

    authors： Beucher M,Meira WS,Zegarra V,Galvão LM,Chiari E,Norris KA

    更新日期：2003-01-01 00:00:00

  • Overexpression of post-translationally modified peptides in Escherichia coli by co-expression with modifying enzymes.

    abstract：:Post-translational modification plays crucial roles in signal transduction in eukaryotic cells. To elucidate the biological function of a protein with a specific post-translational modification, it is necessary to isolate the modified protein. However, it is difficult to incorporate a modified amino acid into a specif...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2007.10.018

    authors： Sugase K,Landes MA,Wright PE,Martinez-Yamout M

    更新日期：2008-02-01 00:00:00

  • A mammalian expression vector for expression and purification of secreted proteins for structural studies.

    abstract：:A mammalian expression vector with features optimized for simple expression and purification of secreted proteins has been developed. This vector was constructed to facilitate X-ray crystallographic studies of cysteine-rich glycoproteins that are difficult to express by other means. Proteins expressed with this vector...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1006/prep.2000.1331

    authors： Leahy DJ,Dann CE 3rd,Longo P,Perman B,Ramyar KX

    更新日期：2000-12-01 00:00:00

  • High-level expression of the Listeria monocytogenes listeriolysin O in Escherichia coli and preliminary characterization of the purified protein.

    abstract：:Listeriolysin O (LLO) is a cholesterol-binding sulfhydryl-activated hemolysin encoded by Listeria monocytogenes hlyA gene. After analyzing the nucleotide coding sequence of this gene from the ATCC 9525 L. monocytogenes strain, we cloned it in a pET vector for expression in Escherichia coli. Thanks to the optimization ...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/s1046-5928(02)00682-4

    authors： Giammarini C,Andreoni F,Amagliani G,Casiere A,Barocci S,Magnani M

    更新日期：2003-03-01 00:00:00

  • Human cell lines: A promising alternative for recombinant FIX production.

    abstract：:Factor IX (FIX) is a vitamin K-dependent protein, and it has become a valuable pharmaceutical in the Hemophilia B treatment. We evaluated the potential of recombinant human FIX (rhFIX) expression in 293T and SK-Hep-1 human cell lines. SK-Hep-1-FIX cells produced higher levels of biologically active protein. The growth...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2015.11.023

    authors： de Sousa Bomfim A,Cristina Corrêa de Freitas M,Picanço-Castro V,de Abreu Soares Neto M,Swiech K,Tadeu Covas D,Maria de Sousa Russo E

    更新日期：2016-05-01 00:00:00

  • Expression of Pisum sativum SAD polypeptides in production hosts and in planta: tetrameric organization of the protein.

    abstract：:In Pisum sativum, the short-chain alcohol dehydrogenase-like protein (SAD) gene family consists of at least three members (SAD-A, -B, and -C). Expression of two of these genes (SAD-A and -C) in Escherichia coli or Pichia pastoris resulted in full-length soluble proteins. Purified SAD-A was used as antigen for antibody...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2008.09.004

    authors： Scherbak N,Brosché M,Ala-Häivälä A,Strid H,Ohrfelt A,Nilsson F,Strid A

    更新日期：2009-01-01 00:00:00

  • Expression, purification, and initial structural characterization of nonstructural protein 2B, an integral membrane protein of Dengue-2 virus, in detergent micelles.

    abstract：:Dengue virus causes serious diseases affecting people in tropical and sub-tropical regions. The nonstructural (NS) protein 2B is an integral membrane protein and important for the regulation of viral protease NS3, which is significant for virus replication. The NS2B-NS3 complex is an important drug target for treating...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2011.08.008

    authors： Huang Q,Chen AS,Li Q,Kang C

    更新日期：2011-12-01 00:00:00

  • Systematic optimization of expression and refolding of the Plasmodium falciparum cysteine protease falcipain-2.

    abstract：:The Plasmodium falciparum cysteine protease falcipain-2 is a potential new target for antimalarial chemotherapy. In order to obtain large quantities of active falcipain-2 for biochemical and structural analysis, a systematic assessment of optimal parameters for the expression and refolding of the protease was carried ...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1006/prep.2001.1416

    authors： Sijwali PS,Brinen LS,Rosenthal PJ

    更新日期：2001-06-01 00:00:00

  • Improving purification of recombinant human interferon gamma expressed in Escherichia coli; effect of removal of impurity on the process yield.

    abstract：:Process development and optimization studies were performed in order to improve the purification process of (rhIFN-gamma). The objective was to generate material with higher purity and quantity. An in-process control screening was developed to obtain the optimal condition for column chromatographic purification by mea...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2006.07.002

    authors： Mohammadian-Mosaabadi J,Naderi-Manesh H,Maghsoudi N,Nassiri-Khalili MA,Masoumian MR,Malek-Sabet N

    更新日期：2007-02-01 00:00:00

  • Use of slow glucose feeding as supporting carbon source in lactose autoinduction medium improves the robustness of protein expression at different aeration conditions.

    abstract：:Recombinant protein expression from lac derived promoters by the autoinduction regime is based on diauxic growth of Escherichia coli on glucose and lactose. Glycerol is used as a supporting carbon source during the lactose-induced expression. While this glycerol-based formulation usually provides high cell densities, ...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2013.07.016

    authors： Ukkonen K,Mayer S,Vasala A,Neubauer P

    更新日期：2013-10-01 00:00:00

  • Enhanced protein expression in the baculovirus/insect cell system using engineered SUMO fusions.

    abstract：:Recombinant protein expression in insect cells varies greatly from protein to protein. A fusion tag that is not only a tool for detection and purification, but also enhances expression and/or solubility would greatly facilitate both structure/function studies and therapeutic protein production. We have shown that fusi...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2008.07.010

    authors： Liu L,Spurrier J,Butt TR,Strickler JE

    更新日期：2008-11-01 00:00:00

  • Confronting high-throughput protein refolding using high pressure and solution screens.

    abstract：:Over-expression of heterologous proteins in Escherichia coli is commonly hindered by the formation of inclusion bodies. Nevertheless, refolding of proteins in vitro has become an essential requirement in the development of structural genomics (proteomics) and as a means of recovering functional proteins from inclusion...

    journal_title：Protein expression and purification

    pub_type： 杂志文章,评审

    doi：10.1016/j.pep.2007.05.014

    authors： Qoronfleh MW,Hesterberg LK,Seefeldt MB

    更新日期：2007-10-01 00:00:00

  • Production of functional human insulin-like growth factor binding proteins (IGFBPs) using recombinant expression in HEK293 cells.

    abstract：:Insulin-like growth factor binding proteins (IGFBPs) display many functions in humans including regulation of the insulin-like growth factor (IGF) signaling pathway. The various roles of human IGFBPs make them attractive protein candidates in drug discovery. Structural and functional knowledge on human proteins with t...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2014.10.017

    authors： Wanscher ASM,Williamson M,Ebersole TW,Streicher W,Wikström M,Cazzamali G

    更新日期：2015-04-01 00:00:00

  • Functional expression, purification, and characterization of human Flt3 ligand in the Pichia pastoris system.

    abstract：:Flt3 ligand (FL) is a potent hematopoietic cytokine that affects the growth and differentiation of hematopoietic progenitor and stem cells both in vivo and in vitro. Pichia pastoris transformants secreting high-level rhFL were obtained using 'yeastern blotting' method and the expression level in liquid was about 30 mg...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2005.04.001

    authors： Zhang YL,Chen SS,Yang KG,Su L,Deng YC,Liu CZ

    更新日期：2005-08-01 00:00:00

  • Expression and purification of the alpha-subunit of eukaryotic initiation factor eIF2: use as a kinase substrate.

    abstract：:The alpha-subunit of eukaryotic initiation factor eIF2 (eIF2alpha) plays an important role in the regulation of mRNA translation through modulation of the interaction of eIF2 and a second initiation factor, eIF2B. The interaction of the two proteins is regulated in vivo by phosphorylation of eIF2alpha at Ser51. In the...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1006/prep.1998.0863

    authors： Kimball SR,Horetsky RL,Jagus R,Jefferson LS

    更新日期：1998-04-01 00:00:00

  • Characterization of homogeneous recombinant glutaredoxin from Escherichia coli: purification from an inducible lambda PL expression system and properties of a novel elongated form.

    abstract：:We have constructed a plasmid, pAHOB1, with a 482-b AluI fragment containing the Escherichia coli glutaredoxin gene (grx) cloned under lambda PL promoter control. Growth of E. coli N4830/pAHOB1 cells at 30 degrees C followed by heat induction at 40 degrees C for 5 h resulted in expression of glutaredoxin as 20% of the...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/1046-5928(91)90085-w

    authors： Björnberg O,Holmgren A

    更新日期：1991-08-01 00:00:00

  • Purification and biochemical characterization of recombinant rat liver phenylalanine hydroxylase produced in Escherichia coli.

    abstract：:Phenylalanine hydroxylase, important in phenylalanine metabolism in mammals, is regulated through short-term (activation) and long-term (induction) mechanisms. To help elucidate the structure-function relationships involved in the activation of this enzyme, we have isolated and characterized full-length cDNA clones to...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：

    authors： Citron BA,Davis MD,Kaufman S

    更新日期：1992-04-01 00:00:00

  • Expression of a soluble and activatable form of bovine procarboxypeptidase A in Escherichia coli.

    abstract：:Bovine pancreatic procarboxypeptidase A has been overexpressed in a soluble and activatable form in Escherichia coli. When the protein was expressed under the control of bacteriophage T7 promoter in E. coli ADA494 (a thioredoxin reductase deficient bacteria), a thioredoxin fusion protein was produced at relatively hig...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/s1046-5928(02)00573-9

    authors： Seddi R,Chaix JC,Puigserver A,Guo XJ

    更新日期：2003-02-01 00:00:00

  • Preparation of soluble isotopically labeled NKp30, a human natural cytotoxicity receptor, for structural studies using NMR.

    abstract：:Using a codon-optimized gene fragment, we report remarkable yields for extracellular domain of human NK cell receptor (NKp30ex) when produced on M9 minimal medium, even with low (2g/L) glucose concentration. The yields were identical using media containing (15)NH(4)Cl or (15)NH(4)Cl in combination with all-(13)C-d-glu...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2012.09.016

    authors： Grave L,Tůmová L,Mrázek H,Kavan D,Chmelík J,Vaněk O,Novák P,Bezouška K

    更新日期：2012-12-01 00:00:00

  • Identification of angiogenesis-inhibiting peptides from Chan Su.

    abstract：:Chan Su is a traditional medicine prepared from toxic secretions from the auricular and skin glands of Chinese toads. Previous studies show that active components in Chan Su can inhibit the proliferation of tumor cells. To study the effect of Chan Su peptides on angiogenesis, fresh Chan Su was collected and its compon...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2019.105445

    authors： Xia F,Gao F,Yao H,Zhang G,Gao B,Lu Y,Wang X,Qian Y

    更新日期：2019-11-01 00:00:00

  • Heterologous expression and purification of plantaricin NC8, a two-peptide bacteriocin against Salmonella spp. from Lactobacillus plantarum ZJ316.

    abstract：:Bacteriocin, which is produced by lactic acid bacteria (LAB), has the potential to act as natural preservatives in the food industry. To develop strategies to overproduce such peptides, plantaricin NC8, a class IIb LAB bacteriocin that consists of two peptides, PLNC8α and PLNC8β, was successfully heterologously expres...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2016.06.013

    authors： Jiang H,Li P,Gu Q

    更新日期：2016-11-01 00:00:00

  • Purification and some properties of human placental glucose-6-phosphate dehydrogenase.

    abstract：:Glucose-6-phosphate dehydrogenase was purified from human placenta using DEAE-Sepharose fast flow, 2',5'-ADP Sepharose 4B column chromatography, and chromatofocusing on PBE 94 with PB 74. The enzyme was purified with 62% yield and had a specific activity of 87 units per milligram protein. The pH optimum was determined...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1006/prep.2000.1370

    authors： Aksoy Y,Ogüs IH,Oauzer N

    更新日期：2001-03-01 00:00:00

  • Recombinant expression of biologically active rat leptin in Escherichia coli.

    abstract：:Leptin is a 16-kDa nonglycosylated hormone that is produced in mature adipocytes and which acts primarily in the hypothalamus to reduce food intake and body weight. While the rat is a representative laboratory animal model in obesity research, so far recombinant rat leptin was not available. In the present study, rat ...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1006/prep.2001.1412

    authors： Park JH,Lee HH,Na SY,Ju SK,Lee YJ,Lee MK,Kim KL

    更新日期：2001-06-01 00:00:00

  • A versatile method for producing labeled or unlabeled Aβ55, Aβ40, and other β-amyloid family peptides.

    abstract：:We present a straightforward, versatile method for expressing and purifying β-amyloid (Aβ40) and transmembrane peptides derived from β-amyloid precursor protein (Aβ55). In principle, these methods should be applicable to other types of strongly aggregating peptides. We start with a DNA plasmid encoding a HexaHis tag w...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2019.04.006

    authors： Zerweck J,Venkata BS,Pittman JM,Srivastava AK,Moore PC,Sachleben JR,Thinakaran G,Meredith SC

    更新日期：2019-10-01 00:00:00

  • Large-scale expression and purification of a soluble form of the pleckstrin homology domain of the human protooncogenic serine/threonine protein kinase PKB (c-akt) in Escherichia coli.

    abstract：:The protooncogenic serine/threonine protein kinase PKB contains an amino-terminal pleckstrin homology (PH) domain which binds phosphatidylinositides. The PH domain, composed of approximately 100 loosely conserved amino acids, is found in many proteins, including kinases, phospholipases C, GTPases, GTPase-activating pr...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1006/prep.1999.1120

    authors： Ingley E,Hemmings BA

    更新日期：1999-11-01 00:00:00

  • High-cell density shake-flask expression and rapid purification of the large fragment of Thermus aquaticus DNA polymerase I using a new chemically and temperature inducible expression plasmid in Escherichia coli.

    abstract：:We have developed a new expression vector, pcI(ts) ind(+), based upon the powerful rightward promoter of bacteriophage lambda, which is controlled by a temperature-sensitive and chemically-inducible version of the lambda repressor on the same plasmid. Locating the repressor gene on the plasmid makes this vector "porta...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2008.09.018

    authors： Brandis JW,Johnson KA

    更新日期：2009-02-01 00:00:00

  • Overexpression and functional characterisation of the human melanocortin 4 receptor in Sf9 cells.

    abstract：:The human melanocortin 4 receptor (MC4r) was successfully expressed in Sf9 cells using the baculovirus infection system. N- and C-terminally His-tagged receptors generated B(max) values of 14 and 23 pmol receptor/mg membrane protein, respectively. The highest expression level obtained with the C-terminally His-tagged ...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2004.06.018

    authors： Dolby V,Collén A,Lundqvist A,Cronet P

    更新日期：2004-10-01 00:00:00

  • Expression, purification, and characterization of deglycosylated human pro-prostate-specific antigen.

    abstract：:Wild-type and deglycosylated forms of human prostate-specific antigen were expressed in Chinese hamster ovary (CHO) cells as zymogens. ProPSA was collected from conditioned medium and purified using a single cation-exchange chromatographic step for the deglycosylated form and cation-exchange followed by gel filtration...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1006/prep.2000.1342

    authors： Bowman KK,Clark J,Yu L,Mortara K,Radika K,Wang J,Zhan H

    更新日期：2000-12-01 00:00:00

  • Functional expression of single-chain Fv antibody in the cytoplasm of Escherichia coli by thioredoxin fusion and co-expression of molecular chaperones.

    abstract：:The production of a single-chain variable fragment (scFv) antibody against bovine ribonuclease A in the cytoplasm of Escherichia coli trxB/gor double mutant was investigated. Previous reports have shown that the thioredoxin (Trx) protein fusion strategy is useful for the correct folding of scFvs and that the expressio...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2009.11.003

    authors： Sonoda H,Kumada Y,Katsuda T,Yamaji H

    更新日期：2010-04-01 00:00:00