Bacterial expression of human kynurenine 3-monooxygenase: solubility, activity, purification.

Abstract:

:Kynurenine 3-monooxygenase (KMO) is an enzyme central to the kynurenine pathway of tryptophan metabolism. KMO has been implicated as a therapeutic target in several disease states, including Huntington's disease. Recombinant human KMO protein production is challenging due to the presence of transmembrane domains, which localise KMO to the outer mitochondrial membrane and render KMO insoluble in many in vitro expression systems. Efficient bacterial expression of human KMO would accelerate drug development of KMO inhibitors but until now this has not been achieved. Here we report the first successful bacterial (Escherichia coli) expression of active FLAG™-tagged human KMO enzyme expressed in the soluble fraction and progress towards its purification.

journal_name

Protein Expr Purif

authors

Wilson K,Mole DJ,Binnie M,Homer NZ,Zheng X,Yard BA,Iredale JP,Auer M,Webster SP

doi

10.1016/j.pep.2013.11.015

subject

Has Abstract

pub_date

2014-03-01 00:00:00

pages

96-103

eissn

1046-5928

issn

1096-0279

pii

S1046-5928(13)00259-3

journal_volume

95

pub_type

杂志文章
  • 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

  • Cloning, overexpression, folding and purification of a biosynthetically derived three disulfide scorpion toxin (BTK-2) from Mesobuthus tamulus.

    abstract::BTK-2, a 32 residue scorpion toxin initially identified in the venom of red Indian scorpion Mesobuthus tamulus was cloned, overexpressed and purified using Cytochrome b(5) fusion protein system developed in our laboratory. The synthetic gene coding for the peptide was designed taking into account optimal codon usage b...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/j.pep.2009.10.017

    authors: Kumar GS,Sarma SP

    更新日期:2010-04-01 00:00:00

  • Unusual stability of manganese superoxide dismutase from a new species, Tatumella ptyseos ct: its gene structure, expression, and enzyme properties.

    abstract::A genomic DNA of 1416 bp containing an open reading frame encoding a manganese superoxide dismutase (Mn-SOD) from Tatumella ptyseos ct was cloned. Sequence analysis of this new gene revealed that it translates 205 amino acid residues. The deduced amino acid sequence showed variable identities (41-91%) with sequences o...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/j.pep.2004.10.003

    authors: Ken CF,Lee CC,Duan KJ,Lin CT

    更新日期:2005-03-01 00:00:00

  • Purification of granulin-like polypeptide from the blood-sucking leech, Hirudo nipponia.

    abstract::A cysteine-rich (approximately 20%), low molecular weight (MW 6 kDa) polypeptide has been isolated from the Korean blood-sucking leech, Hirudo nipponia. From its amino acid composition and N-terminal amino acid sequence analysis, the new protein is similar to granulin (or epithelin), and so it has been named leech gra...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1006/prep.1999.1077

    authors: Hong SJ,Kang KW

    更新日期:1999-07-01 00:00:00

  • A simple and efficient method for generating high-quality recombinant Mical enzyme for in vitro assays.

    abstract::We have recently identified a new family of multidomain oxidoreductase (redox) enzymes, the MICALs, that directly regulate the actin cytoskeletal elements necessary for the morphology, motility, and trajectory of cells. Our genetic assays reveal that Mical is both necessary and sufficient for actin organization and ce...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/j.pep.2016.05.008

    authors: Wu H,Hung RJ,Terman JR

    更新日期:2016-11-01 00:00:00

  • Expression and reconstitution of NF-kappaB from insect cells using a baculovirus vector.

    abstract::NF-kappaB is a pleiotropic transcriptional activator originally identified by its ability to regulate immunoglogulin kappa light chain expression. Purification of this DNA-binding complex demonstrated that NF-kappaB is a heterodimer composed of two subunits, NFKB1 and RelA. Previous studies have shown that truncated v...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1006/prep.1996.0670

    authors: Coleman TA,Huddleston KA,Ruben SM,Rosen CA,Gentz R

    更新日期:1997-02-01 00:00:00

  • Expression of the recombinant antibacterial peptide sarcotoxin IA in Escherichia coli cells.

    abstract::Sarcotoxin IA is an antibacterial peptide that is secreted by a meat-fly Sarcophaga peregrina larva in response to a hypodermic injury or bacterial infection. This peptide is highly toxic against a broad spectrum of both Gram-positive and Gram-negative bacteria and lethal to microbes even at nanomolar concentrations. ...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/s1046-5928(02)00697-6

    authors: Skosyrev VS,Kulesskiy EA,Yakhnin AV,Temirov YV,Vinokurov LM

    更新日期:2003-04-01 00:00:00

  • A three-step purification of manganese superoxide dismutase from human liver on both large and small scales.

    abstract::A new method for the purification of manganese superoxide dismutase from human liver is described. The procedure involves essentially three steps: DEAE-cellulose, hydroxylapatite, and butyl-Toyopearl chromatographies. The method has several advantages: (i) its simplicity and rapidity (it takes less than 3 days), (ii) ...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/1046-5928(91)90067-s

    authors: Matsuda Y,Suzuki K,Ookawara T,Nakata T,Seo HG,Kawata S,Tarui S,Deutsch HF,Taniguchi N

    更新日期:1991-04-01 00:00:00

  • Method for efficient soluble expression and purification of recombinant human interleukin-15.

    abstract::Periplasmic expression of recombinant proteins ensures the production of biologically active proteins in a correctly folded state with several key advantages. This research focused on the in-frame cloning of rhIL-15 in pET-20 (+) vector with pelB-leader sequence to direct the protein to the bacterial periplasm. The ta...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/j.pep.2020.105746

    authors: Ahmed N,Afroze B,Abbas R,Khan MA,Akram M,Tahir S,Bakht S,Munir A,Shahid AA

    更新日期:2021-01-01 00:00:00

  • Cloning, expression, purification, and characterization of Nocardia sp. GTP cyclohydrolase I.

    abstract::The sequence of the gene from Nocardia sp. NRRL 5646 encoding GTP cyclohydrolase I (GCH), gch, and its adjacent regions was determined. The open reading frame of Nocardia gch contains 684 nucleotides, and the deduced amino acid sequence represents a protein of 227 amino acid residues with a calculated molecular mass o...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/j.pep.2004.02.008

    authors: He A,Simpson DR,Daniels L,Rosazza JP

    更新日期:2004-06-01 00:00:00

  • Affinity purification of recombinant human cytochrome P450s 3A4 and 1A2 using mixed micelle systems.

    abstract::Recombinant cytochrome P450 (CYP or P450) enzymes are useful for drug metabolism research and thereby many expression and purification systems have been developed. Here, we provide a method for the purification of human P450s 3A4 and 1A2 expressed in Escherichia coli using mixed micelles containing anionic phospholipi...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/j.pep.2014.05.010

    authors: Ahn T,Bae CS,Yun CH

    更新日期:2014-09-01 00:00:00

  • Overexpression and purification of the Escherichia coli inner membrane enzyme acyl-acyl carrier protein synthase in an active form.

    abstract::Acyl-acyl carrier protein synthase (Aas) is widely used to synthesize thioester adducts of fatty acids between 8 and 18 carbons in length enzymatically to the phosphopantetheine group of acyl carrier protein. The enzyme is an 80.6-kDa inner membrane protein that functions in vivo as a 2-acylglycerophosphoethanolamine ...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1006/prep.2000.1206

    authors: Shanklin J

    更新日期:2000-04-01 00:00:00

  • Expression and purification of receptor for activated C-kinase 1 (RACK1).

    abstract::Receptor for activated C-kinase (RACK1) binds to protein kinase C and functions as an anchor for several other cellular components. Most in vitro studies of RACK1 have been carried out with RACK1 fused to a soluble fusion protein partner, such as GST or MBP. Here, we show that fusion complexes may exist as large solub...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/s1046-5928(03)00135-9

    authors: Bjørndal B,Trave G,Hageberg I,Lillehaug JR,Raae AJ

    更新日期:2003-09-01 00:00:00

  • Functional recombinant rabbit muscle phosphoglucomutase from Escherichia coli.

    abstract::The gene coding for phosphoglucomutase (PGM) from Oryctolagus cuniculus (rabbit) has been expressed in Escherichia coli under a T7 expression system with a His-tag. About half of the expressed PGM protein was present in inclusion bodies, but this protein was inactive when solubilized. The protein in the soluble cell f...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1006/prep.2000.1288

    authors: Chae YK,Markley JL

    更新日期:2000-10-01 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

  • A new method for the purification of bioactive insulin-like growth factor-binding protein-3.

    abstract::We present a novel efficient procedure for high level purification of human IGFBP-3. Insulin-like growth factor-binding proteins (IGFBPs) are key regulators of insulin-like growth factor mediated signal transduction and thereby can profoundly influence cellular phenotypes. Certain IGFBPs, including IGFBP-3, have also ...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/j.pep.2010.02.005

    authors: Pircher H,Matscheski A,Laich A,Hermann M,Moser B,Viertler HP,Micutkova L,Lindner H,Sarg B,Zwerschke W,Jansen-Dürr P

    更新日期:2010-06-01 00:00:00

  • One-step affinity tag purification of full-length recombinant human AP-1 complexes from bacterial inclusion bodies using a polycistronic expression system.

    abstract::The AP-1 transcription factor is a dimeric protein complex formed primarily between Jun (c-Jun, JunB, JunD) and Fos (c-Fos, FosB, Fra-1, Fra-2) family members. These distinct AP-1 complexes are expressed in many cell types and modulate target gene expression implicated in cell proliferation, differentiation, and stres...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/j.pep.2008.01.016

    authors: Wang WM,Lee AY,Chiang CM

    更新日期:2008-05-01 00:00:00

  • Purification and characterization of a soybean root nodule phosphatase expressed in Pichia pastoris.

    abstract::Soybean root nodules possess a developmentally regulated acid phosphatase (ACP) that exhibits the highest specificity for purine 5'-nucleoside monophosphates. The enzyme is a glycosylated dimer of 28- and 31-kDa subunits, which appear to be products of the same gene but differ in posttranslational modifications. In or...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1006/prep.1998.0935

    authors: Penheiter AR,Klucas RV,Sarath G

    更新日期:1998-10-01 00:00:00

  • Affinity chromatographic purification of a protein which binds specifically to the yeast leucine tRNA gene.

    abstract::A crude cell extract from yeast Saccharomyces cerevisae was fractionated by affinity chromatography using the leucine tRNA gene as the recognition site. This approach enables the rapid purification of a protein, which retained its full DNA binding capacity during the enrichment procedure. The active fraction contains ...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/1046-5928(90)90015-q

    authors: Kaufmann E

    更新日期:1990-11-01 00:00:00

  • Expression and purification of recombinant human tyrosine hydroxylase as a fusion protein in Escherichia coli.

    abstract::Tyrosine hydroxylase is the rate-limiting step in the synthesis of dopamine and is tightly regulated. Previous studies have shown it to be covalently modified and potently inhibited by 3,4-dihydroxyphenylacetaldehyde (DOPAL), an endogenous neurotoxin via dopamine catabolism which is relevant to Parkinson's disease. In...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/j.pep.2012.05.007

    authors: Higgins CA,Vermeer LM,Doorn JA,Roman DL

    更新日期:2012-08-01 00:00:00

  • Expression of a functional cold active β-galactosidase from Planococcus sp-L4 in Pichia pastoris.

    abstract::Lactase deficiency problem discourages many adults from consuming milk as a major source of micro- and macronutrients. Enzymatic hydrolysis of lactose is an ideal solution for this problem but such processing adds significant costs. In this study, a cold active β-galactosidase from Planococcus sp-L4 (bgal) was optimiz...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/j.pep.2015.09.008

    authors: Mahdian SM,Karimi E,Tanipour MH,Parizadeh SM,Ghayour-Mobarhan M,Bazaz MM,Mashkani B

    更新日期:2016-09-01 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

  • Expression, purification and structural properties of ABC transporter ABCA4 and its individual domains.

    abstract::ABCA4 is a member of the A subfamily of ATP-binding cassette transporters that consists of large integral membrane proteins implicated in inherited human diseases. ABCA4 assists in the clearance of N-retinylidene-phosphatidylethanolamine, a potentially toxic by-product of the visual cycle formed in photoreceptor cells...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/j.pep.2014.02.010

    authors: Tsybovsky Y,Palczewski K

    更新日期:2014-05-01 00:00:00

  • Expression in Escherichia coli: purification and characterization of cyclin H, a subunit of the human general transcription/DNA repair factor TFIIH.

    abstract::The human cyclin H, a protein normally associated with the cyclin-dependent kinase cdk7, was overexpressed in Escherichia coli using a T7 RNA polymerase expression system and further purified to apparent homogeneity. The purified recombinant cyclin H is similar to the endogenous one according to the following criteria...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1006/prep.1996.0693

    authors: Poterszman A,Andersen G,Busso D,Rossignol M,Egly JM,Thierry JC

    更新日期:1997-03-01 00:00:00

  • Purification and characterization of a cold-adapted κ-carrageenase from Pseudoalteromonas sp. ZDY3.

    abstract::κ-Carrageenase (EC3.2.1.83) is a class of glycoside hydrolase, which can be used for hydrolysis of κ-carrageenan to κ-carrageenan oligosaccharides. In this study, a bacterium, identified as Pseudoalteromonas sp. ZDY3 isolated from rotten algae, was capable to degrade κ-carrageenan. The κ-carrageenase produced by Pseud...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/j.pep.2020.105768

    authors: Zhao D,Jiang B,Zhang Y,Sun W,Pu Z,Bao Y

    更新日期:2021-02-01 00:00:00

  • Cloning and constitutive expression of His-tagged xylanase GH 11 from Penicillium occitanis Pol6 in Pichia pastoris X33: purification and characterization.

    abstract::High-level constitutive expression of xylanase GH11 from Penicillium occitanis Pol6 termed PoXyn2 was achieved using the methylotrophic yeast Pichia pastoris. The PoXyn2 cDNA encoding for a mature xylanase of 320 amino acids was subcloned into the pGAPZαA vector, to construct recombinant xylanse with six histidine res...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/j.pep.2012.02.012

    authors: Driss D,Bhiri F,Ghorbel R,Chaabouni SE

    更新日期:2012-05-01 00:00:00

  • Bacterial fermentation of recombinant major wasp allergen Antigen 5 using oxygen limiting growth conditions improves yield and quality of inclusion bodies.

    abstract::A process for bacterial expression and purification of the recombinant major wasp allergen Antigen 5 (Ves v 5) was developed to produce protein for diagnostic and therapeutic applications for type 1 allergic diseases. Special attention was focused on medium selection, fermentation conditions, and efficient refolding p...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/j.pep.2006.01.009

    authors: Kischnick S,Weber B,Verdino P,Keller W,Sanders EA,Anspach FB,Fiebig H,Cromwell O,Suck R

    更新日期:2006-06-01 00:00:00

  • Purification of a maize dehydrin.

    abstract::A maize dehydrin with an apparent molecular weight of 20 kDa was purified from whole kernels of maize inbred line G50. Kernels were ground in a seed mill, stirred overnight in extraction buffer, and centrifuged to extract soluble proteins. The sample was heated to 89 degrees C and centrifuged to remove heat-insoluble ...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1006/prep.1994.1040

    authors: Ceccardi TL,Meyer NC,Close TJ

    更新日期:1994-06-01 00:00:00

  • Improved PC1/3 production through recombinant expression in insect cells and larvae.

    abstract::Protein convertase 1/3 is a serine endoproteinase present in the regulated secretory pathway of endocrine and neuroendocrine cells. It is responsible for the processing of numerous prohormones and proneuropeptides into their biologically active moieties, often following cleavage at pairs of basic residues. The determi...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/j.pep.2004.06.014

    authors: Rabah N,Gauthier DJ,Gauthier D,Lazure C

    更新日期:2004-10-01 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