Expression and purification of human antimicrobial peptide, dermcidin, in Escherichia coli.

Abstract:

:Human dermcidin, an anionic antimicrobial peptide expressed in the pons of the brain and the sweat glands, displays antimicrobial activity against pathogenic microorganisms such as Staphylococcus aureus and Candida albicans. Here, we describe the recombinant production of a 48 amino acid dermcidin variant with C-terminal homoserine lactone (DCD-1Hsl). Dermcidin coding sequence was cloned downstream of a 125 amino acid ketosteroid isomerase gene and upstream of a His6Tag sequence in pET-31b(+) vector and transformed into Escherichia coli. The fusion protein was expressed in the form of inclusion bodies, purified on His Bind Resin, and cleaved by CNBr to release recombinant DCD-1Hsl. Purification of rDCD-1Hsl was achieved by solid-phase extraction that yielded milligram amounts of peptide with more than 95% purity. Recombinant peptide showed antimicrobial activities against E. coli ML-35p, Salmonella typhimurium 5156, Listeria monocytogenes 264, S. aureus 29/58 (clinical isolate), and C. albicans K2 (clinical strain). The application of this expression/purification approach represents a fast and efficient method to prepare milligram quantities of dermcidin in its biologically active form.

journal_name

Protein Expr Purif

authors

Cipáková I,Gasperík J,Hostinová E

doi

10.1016/j.pep.2005.07.002

keywords:

subject

Has Abstract

pub_date

2006-02-01 00:00:00

pages

269-74

issue

2

eissn

1046-5928

issn

1096-0279

pii

S1046-5928(05)00229-9

journal_volume

45

pub_type

杂志文章
  • Kinetic characterization of recombinant human cystathionine beta-synthase purified from E. coli.

    abstract::Cystathionine beta-synthase (CBS) catalyzes the pyridoxal-50-phosphate-dependent condensation of L-serine and L-homocysteine to form L-cystathionine in the first step of the transsulfuration pathway. Although effective expression systems for recombinant human CBS (hCBS) have been developed, they require multiple chrom...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/j.pep.2008.10.012

    authors: Belew MS,Quazi FI,Willmore WG,Aitken SM

    更新日期:2009-04-01 00:00:00

  • Secretory expression and purification of Aspergillus niger glucose oxidase in Saccharomyces cerevisiae mutant deficient in PMR1 gene.

    abstract::The gene encoding glucose oxidase (GOD) from Aspergillus niger was expressed as a secretory product in the yeast Saccharomyces cerevisiae. Six consecutive histidine residues were fused to the C-terminus of GOD to facilitate purification. The recombinant GOD-His(6) secreted by S. cerevisiae migrated as a broad diffuse ...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/s1046-5928(02)00035-9

    authors: Ko JH,Hahm MS,Kang HA,Nam SW,Chung BH

    更新日期:2002-08-01 00:00:00

  • Isolation of alpha-lactalbumin, beta-lactoglobulin, and bovine serum albumin from cow's milk using gel filtration and anion-exchange chromatography including evaluation of their antigenicity.

    abstract::The aim of this study was to introduce a simple, reproducible, and less expensive method for isolation of alpha-lactalbumin, beta-lactoglobulin, and bovine serum albumin from cow's milk while retaining their antigenicity. Whey (lactoserum) was obtained by isolating casein from defatted milk using hydrochloric acid. Gl...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/s1046-5928(03)00015-9

    authors: Neyestani TR,Djalali M,Pezeshki M

    更新日期:2003-06-01 00:00:00

  • Refolding and one-step purification of recombinant human ARA70 over-expressed in Escherichia coli.

    abstract::Androgen receptor (AR)-associated coregulator 70 (ARA70) is a cytoplasmic protein that has been characterized to have the ability to induce AR transcriptional activity in response to androgens and anti-androgens in prostate cancer cells. AR has been shown to have an important role in the progression of prostate cancer...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/j.pep.2004.11.002

    authors: Singh VK,Jia Z

    更新日期:2005-02-01 00:00:00

  • Secretory production and purification of functional full-length streptavidin from Bacillus subtilis.

    abstract::Streptavidin is a versatile molecule for many in vitro and in vivo applications. To optimize the production of the full-length streptavidin in a soluble and functional form via secretion using Bacillus subtilis as the expression host, three different strategies were used. These strategies include the construction of a...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1006/prep.2001.1582

    authors: Wu SC,Hassan Qureshi M,Wong SL

    更新日期:2002-04-01 00:00:00

  • The identification and expression of the full-length HtrA2 gene from Penaeus monodon (black tiger shrimp).

    abstract::HtrA2 is an apoptosis-activating protein to enhance the apoptotic process by preventing the formation of the IAP-caspase complex, thus freeing caspase to trigger the apoptosis pathway. Here, we presented the full-length sequence of HtrA2 from the black tiger shrimp (PmHtrA2). The full-length PmHtrA2 transcript was 140...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/j.pep.2013.09.012

    authors: Suwannaboon R,Phiwsaiya K,Senapin S,Khunrae P,Rattanarojpong T

    更新日期:2013-12-01 00:00:00

  • Production and secretion of Lactobacillus crispatus β-galactosidase in Pichia pastoris.

    abstract::Lactobacillus β-galactosidases are mostly heterodimeric proteins, which are encoded by the two overlapping genes, lacL and lacM, and produced in recombinant prokaryotic systems for higher yield. This is the first report on the expression of a heterodimeric β-galactosidase from Lactobacillus crispatus B470 in Pichia pa...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/j.pep.2013.08.019

    authors: Nie C,Liu B,Zhang Y,Zhao G,Fan X,Ning X,Zhang W

    更新日期:2013-11-01 00:00:00

  • Heterologous expression of Class IIb bacteriocin Plantaricin JK in Lactococcus Lactis.

    abstract::Plantaricin JK (PlnJK) is a Class IIb LAB bacteriocin that includes two peptides; i.e., PlnJ and PlnK, which can synergistically halt many types of gram-positive bacteria, including food spoilage organisms. Purification of these peptides from natural lactic acid bacteria is difficult therefore, their application remai...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/j.pep.2019.02.013

    authors: Xu Y,Yang L,Li P,Gu Q

    更新日期:2019-07-01 00:00:00

  • High-throughput proteomics: protein expression and purification in the postgenomic world.

    abstract::Proteomics has become a major focus as researchers attempt to understand the vast amount of genomic information. Protein complexity makes identifying and understanding gene function inherently difficult. The challenge of studying proteins in a global way is driving the development of new technologies for systematic an...

    journal_title:Protein expression and purification

    pub_type: 杂志文章,评审

    doi:10.1006/prep.2001.1465

    authors: Lesley SA

    更新日期:2001-07-01 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

  • An alternative use of basic pGEX vectors for producing both N- and C-terminal fusion proteins for production and affinity purification of antibodies.

    abstract::Glutathione S-transferase (GST) fusion proteins are widely used in protein production for pure immunogens, protein-protein, and DNA-protein interaction studies. Using basic pGEX vectors, foreign DNA is introduced to the C-terminus of the GST gene and the produced fusion proteins are C-terminally orientated. However, b...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/j.pep.2004.11.012

    authors: Aatsinki JT,Rajaniemi HJ

    更新日期:2005-04-01 00:00:00

  • Statistical approaches to maximize recombinant protein expression in Escherichia coli: a general review.

    abstract::The supply of many valuable proteins that have potential clinical or industrial use is often limited by their low natural availability. With the modern advances in genomics, proteomics and bioinformatics, the number of proteins being produced using recombinant techniques is exponentially increasing and seems to guaran...

    journal_title:Protein expression and purification

    pub_type: 杂志文章,评审

    doi:10.1016/j.pep.2013.10.016

    authors: Papaneophytou CP,Kontopidis G

    更新日期:2014-02-01 00:00:00

  • "CodonWizard" - An intuitive software tool with graphical user interface for customizable codon optimization in protein expression efforts.

    abstract::Optimization of coding sequences to maximize protein expression yield is often outsourced to external service providers during commercial gene synthesis and thus unfortunately remains a black box for many researchers. The presented software program "CodonWizard" offers scientists a powerful but easy-to-use tool for cu...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/j.pep.2019.03.018

    authors: Rehbein P,Berz J,Kreisel P,Schwalbe H

    更新日期:2019-08-01 00:00:00

  • Engineering S-protein fragments of bovine ribonuclease A for targeted drug delivery.

    abstract::High affinity interaction between S-protein and S-peptide fragments of bovine pancreatic RNase A has been recently used for construction of molecular vehicles for targeted drug delivery. The vehicle is assembled as a complex of drug carrier conjugated S-protein with S-peptide-tagged targeting protein. To avoid random ...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/s1046-5928(02)00546-6

    authors: Backer MV,Gaynutdinov TI,Aloise R,Przekop K,Backer JM

    更新日期:2002-12-01 00:00:00

  • Baculovirus expression and purification of a soluble, mutant G-protein alpha subunit.

    abstract::The cDNA for the alpha i1 protein that had undergone site-directed mutagenesis to change glycine-2 to alanine was ligated into a baculovirus transfer vector. A recombinant virus was obtained by transfecting Sf9 cells with both the wild-type baculovirus DNA and the transfer vector and screening for recombinant plaques....

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1006/prep.1993.1010

    authors: Jones TL,Woodard C,Spiegel AM

    更新日期:1993-02-01 00:00:00

  • Expression, purification, and functional analysis of the TyrR protein of Haemophilus influenzae.

    abstract::The gene that was inferred to encode the TyrR protein of Haemophilus influenzae Rd was synthesized by polymerase chain reaction and inserted into a T7-based expression vector. Methods were developed to overexpress the TyrR protein of H. influenzae in Escherichia coli and to purify the protein on a large scale. Both in...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1006/prep.1997.0757

    authors: Zhu Q,Zhao S,Somerville RL

    更新日期:1997-07-01 00:00:00

  • Protein purification with C-terminal fusion of maltose binding protein.

    abstract::For affinity-chromatography-based purification of proteins that are prone to abnormal termination of translation or that may not be modified at their N-termini, affinity tags are needed which can be fused to the C-terminus. In this publication we describe that maltose binding protein (MBP) fused to the C-terminus of t...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1006/prep.1998.0969

    authors: Hennig L,Schäfer E

    更新日期:1998-12-01 00:00:00

  • Analysis of human alpha-thrombin by hydrophobic interaction high-performance liquid chromatography.

    abstract::The major active form of human thrombin, alpha-thrombin, was analyzed by hydrophobic interaction high-performance liquid chromatography (HIC-HPLC). The chromatographic system included a polymeric phenyl column and elution was performed by a gradient, 2-0M sodium chloride (5-20 min). Total analysis time was 30 min per ...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/s1046-5928(02)00596-x

    authors: Karlsson G

    更新日期:2003-01-01 00:00:00

  • Expression and properties of arginyl-tRNA synthetase from jack bean (Canavalia ensiformis).

    abstract::The coding region for arginyl-tRNA synthetase from jack bean (Canavalia ensiformis) has been sequenced and cloned into the bacterial expression vector pET32a. Transformation of BL21 cells and induction with IPTG results in the high level expression of the protein fused N-terminally with thioredoxin and bearing a His-t...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/j.pep.2008.05.008

    authors: Hogg J,Schiefermayr E,Schiltz E,Igloi GL

    更新日期:2008-10-01 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

  • 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

  • Expression, purification, and refolding of biologically active Acinetobacter baumannii OmpA from Escherichia coli inclusion bodies.

    abstract::Infections caused by Acinetobacter baumannii have emerged as a significant clinical problem due to the increase in infections caused by antibiotic resistant strains. A. baumannii OmpA is a highly conserved membrane protein that has multiple roles in interacting with the host during infection, and thus represents an at...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/j.pep.2010.11.019

    authors: McConnell MJ,Pachón J

    更新日期:2011-05-01 00:00:00

  • The human rhabdomyosarcoma cell line TE671--Towards an innovative production platform for glycosylated biopharmaceuticals.

    abstract::The market of therapeutic glycoproteins (including coagulation factors, antibodies, cytokines and hormones) is one of the profitable, fast-growing and challenging sectors of the biopharmaceutical industry. Although mammalian cell culture is still expensive and technically complex, the ability to produce desired post-t...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/j.pep.2015.08.008

    authors: Rosenlöcher J,Weilandt C,Sandig G,Reinke SO,Blanchard V,Hinderlich S

    更新日期:2015-11-01 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

  • Practical considerations in refolding proteins from inclusion bodies.

    abstract::Refolding of proteins from inclusion bodies is affected by several factors, including solubilization of inclusion bodies by denaturants, removal of the denaturant, and assistance of refolding by small molecule additives. We will review key parameters associated with (1) conformation of the protein solubilized from inc...

    journal_title:Protein expression and purification

    pub_type: 杂志文章,评审

    doi:10.1016/s1046-5928(02)00641-1

    authors: Tsumoto K,Ejima D,Kumagai I,Arakawa T

    更新日期:2003-03-01 00:00:00

  • Hepatocyte growth factor (HGF) is a copper-binding protein: a facile probe for purification of HGF by immobilized Cu(II)-affinity chromatography.

    abstract::Hepatocyte growth factor (HGF) is a multifunctional protein expressed in a variety of cell types and tissues. Here we describe a novel one-step method to separate and identify HGF, based on a unique interaction between HGF and Cu(II). Conditioned medium (CM) from mouse 3T3-L1 adipocytes which contains HGF or purified ...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1006/prep.1996.0047

    authors: Rahimi N,Etchells S,Elliott B

    更新日期:1996-05-01 00:00:00

  • Expression and purification of codon-optimized cre recombinase in E. coli.

    abstract::The presence of antibiotic resistance genes in genetically modified bacteria raises a regulatory concern in the production of therapeutic proteins and additionally reduces the number of plasmids available for propagation in a cell. Cre recombinase from bacteriophage P1, involved in Cre/loxP mechanism is one of the wid...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/j.pep.2019.105546

    authors: D S,Shyam Mohan AH,Rao SN

    更新日期:2020-03-01 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

  • Refolding, characterization, and dye decolorization ability of a highly thermostable laccase from Geobacillus sp. JS12.

    abstract::A putative laccase gene (lacG) from Geobacillus sp. JS12 was cloned and expressed as a fusion protein with six histidine residues in Escherichia coli BL21 (DE3) cells, and the protein was primarily found in inclusion bodies. The resulting insoluble proteins were solubilized with 6 M guanidine HCl and refolded using an...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/j.pep.2020.105646

    authors: Jeon SJ,Park JH

    更新日期:2020-09-01 00:00:00

  • Bax to the future - A novel, high-yielding approach for purification and expression of full-length Bax protein for structural studies.

    abstract::Mitochondria-mediated apoptosis (programmed cell death) involves a sophisticated signaling and regulatory network that is regulated by the Bcl-2 protein family. Members of this family have either pro- or anti-apoptotic functions. An important pro-apoptotic member of this family is the cytosolic Bax. This protein is cr...

    journal_title:Protein expression and purification

    pub_type: 杂志文章

    doi:10.1016/j.pep.2019.02.004

    authors: Dingeldein APG,Lindberg MJ,Ådén J,Zhong X,Stoll R,Gröbner G

    更新日期:2019-06-01 00:00:00