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 of Mn-SODs from other species. The residues required to coordinate the single trivalent manganese ion and the 11 residues putatively involved in the active center are conserved as they are in other reported Mn-SODs. In addition, the gene was introduced into the expression vector, pET-20b(+), and transformed in Escherichia coli BL21(DE3). The Mn-SOD was purified by a His-tag technique. The yield was 0.9 mg from 0.5 L of culture. The specific activity was 6540 U/mg. A dimer is the major form of the enzyme in equilibrium. The half-life of dimer is approximately 50 min and its thermal inactivation rate constant k(d) was 0.015 min(-1) at 80 degrees C. The dimerization of the enzyme was inhibited under an acidic pH (below 4.0), or in the presence of SDS (above 1%) or imidazole (above 0.5 M), whereas it was not affected under an alkaline pH (above 9.0). Furthermore, the dimeric enzyme was much more resistant to proteolytic attack after 3 h of incubation at 37 degrees C with trypsin and chymotrypsin. This unusually stable enzyme can be used as cosmetic to the protection of skin against the unaesthetic effects caused by free radicals.
journal_name
Protein Expr Purifjournal_title
Protein expression and purificationauthors
Ken CF,Lee CC,Duan KJ,Lin CTdoi
10.1016/j.pep.2004.10.003keywords:
subject
Has Abstractpub_date
2005-03-01 00:00:00pages
42-50issue
1eissn
1046-5928issn
1096-0279pii
S1046-5928(04)00336-5journal_volume
40pub_type
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journal_title:Protein expression and purification
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journal_title:Protein expression and purification
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journal_title:Protein expression and purification
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journal_title:Protein expression and purification
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doi:10.1016/1046-5928(92)90005-h
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abstract::Polyhistidine tags enable the facile purification of proteins by immobilized metal affinity chromatography (IMAC). Both the type and position of purification tags can affect significantly properties of a protein such as its expression level, behavior in solution, and its ability to form suitable samples (esp. suitable...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2003.10.010
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abstract::Introduction and expression of foreign genes in bacteria often results accumulation of the foreign protein(s) in inclusion bodies (IBs). The subsequent processes of refolding are slow, difficult and often fail to yield significant amounts of folded protein. RHG1 encoded by rhg1 was a soybean (Glycine max L. Merr.) tra...
journal_title:Protein expression and purification
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journal_title:Protein expression and purification
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abstract::In this report, we describe an optimized system for the efficient overexpression, purification, and refolding of secreted bacterial proteins. Candidate secreted proteins were produced recombinantly in Escherichia coli as Tobacco Etch Virus protease-cleavable hexahistidine-c-myc eptiope fusion proteins. Without regard ...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2005.09.003
更新日期:2006-03-01 00:00:00
abstract::A fibrinolytic enzyme was purified from the cultured mycelia of Armillaria mellea by ion-exchange chromatography followed by gel filtration, and was designated A. mellea metalloprotease (AMMP). The purification protocol resulted in a 627-fold purification of the enzyme, with a final yield of 6.05%. The apparent molecu...
journal_title:Protein expression and purification
pub_type: 杂志文章
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journal_title:Protein expression and purification
pub_type: 杂志文章
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abstract::Recombinant baculoviruses have proved to be a very useful means to express many proteins over the last 20 years. Since their introduction, there have been a number of significant improvements that have simplified and speeded up the construction of baculoviruses. One of the most commonly used methods relies upon recomb...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2005.03.021
更新日期:2005-07-01 00:00:00
abstract::p53 protein is an important regulation factor that can bind to p53 mRNA to regulate its translation in human and murine. To determine if a similar interaction exists in zebrafish and if the interaction affects zebrafish development, we cloned and expressed p53 protein from zebrafish in Escherichia coli. Soluble p53 pr...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2010.03.028
更新日期:2010-08-01 00:00:00
abstract::Molecular chaperones are integral components of the cellular machinery involved in ensuring correct protein folding and the continued maintenance of protein structure. An understanding of these ubiquitous molecules is key to finding cures to protein misfolding diseases such as Alzheimer's and Creutzfeldt-Jacob disease...
journal_title:Protein expression and purification
pub_type: 杂志文章,评审
doi:10.1016/j.pep.2005.08.005
更新日期:2006-03-01 00:00:00
abstract::Human Relaxin 2 is an insulin-related peptide hormone with a mass of 19,084 Da. The mRNA contains a number of arginine codons that are rarely used by Escherichia coli to produce highly expressed proteins. As a result, expressing this recombinant protein in E. coli is problematic. When human Relaxin 2 was expressed in ...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2008.02.016
更新日期:2008-08-01 00:00:00
abstract::Proteus mirabilis (P. mirabilis) is a zoonotic pathogen that has recently presented a rising infection rate in the poultry industry. To develop an effective vaccine to protect chickens against P. mirabilis infection, OmpA, one of the major outer membrane proteins of P. mirabilis, was expressed in Pichia pastoris. The ...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2014.10.001
更新日期:2015-01-01 00:00:00
abstract::The fusion protein consisting of human Fas receptor extracellular domain and human IgG1 heavy chain Fc domain (hFasRECD-Fc) is a medically important protein that potentially has therapeutic uses. The fusion gene composed of a synthetic human Fas receptor extracellular domain gene and the cDNA encoding human IgG1 heavy...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2010.05.007
更新日期:2010-10-01 00:00:00
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
更新日期:1999-07-01 00:00:00
abstract::Glucose-6-phosphate dehydrogenase (G6PDH) (EC 1.1.1.363) plays an important role in the human pathogen Pseudomonas aeruginosa because it generates NADPH, an essential cofactor for several biosynthetic pathways and antioxidant enzymes. P. aeruginosa G6PDH is also a key enzyme in the metabolism of various carbon sources...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2017.10.004
更新日期:2018-02-01 00:00:00
abstract::Enteropeptidase (synonym: enterokinase, EC 3.4.21.9) is a heterodimeric serine protease of the intestinal brush border that activates trypsinogen by highly specific cleavage of the trypsinogen activation peptide following the sequence (Asp)(4)-Lys. It has also great biotechnological interest because of the unique subs...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2011.04.005
更新日期:2011-10-01 00:00:00
abstract::The immunosuppressive drug FK506 binds its targets FK506-binding protein (FKBP) family and modulates cellular processes. Recent studies demonstrated that FK506 shows anti-malaria effects. Newly identified FK506-binding protein 35 from Plasmodium falciparum (PfFKBP35) is assumed to be the molecular target of FK506 in t...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2006.12.019
更新日期:2007-05-01 00:00:00
abstract::Silent information regulator 2 (Sir2) proteins are a class of protein deacetylase enzymes that play key roles in transcriptional gene silencing, DNA repair, and aging. Here, we describe the high-level bacterial expression and purification of a human SirT2 construct that yields high resolution NMR spectra. By removing ...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2005.12.006
更新日期:2006-07-01 00:00:00
abstract::Hsp47 is regarded as a collagen-specific chaperone with several suggested roles in collagen biosynthesis under normal and disease conditions. We describe here a procedure for the expression and purification of Hsp47 in Escherichia coli using the IMPACT expression system (New England Biolabs) where the guest gene is fu...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.2001.1470
更新日期:2001-10-01 00:00:00
abstract::Human lysosomal beta-hexosaminidase exists in two major forms: the A isoform is composed of both alpha and beta chains, while the B form is a homopolymer of beta chains. Deficiency of beta-hexosaminidase underlies the GM2 gangliosidoses. We have produced active beta-hexosaminidase B in cultured insect (Sf9) cells by i...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/1046-5928(90)90003-h
更新日期:1990-11-01 00:00:00
abstract::The Human Secretome Project aims to produce and purify all human secreted proteins as full-length. In order to enable this, a robust, gentle and effective purification process is needed, where multiple proteins can be purified in parallel. For this reason, a purification system based on a Protein C-tag and the HPC4 an...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2020.105698
更新日期:2020-11-01 00:00:00
abstract::Virus-like particles (VLPs) of the recombinant hepatitis B virus (HBV) core protein (HBc) are routinely used in HBV diagnostics worldwide and are of potential interest as carriers of foreign peptides (e.g., immunological epitopes and targeting addresses, and/or as vessels for packaged diagnostic and therapeutic nanoma...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2010.09.010
更新日期:2011-02-01 00:00:00
abstract::The P(II) proteins comprise a family of widely distributed signal transduction proteins that integrate the signals of cellular nitrogen, carbon and energy status, and then regulate, by protein-protein interaction, the activity of a variety of target proteins including enzymes, transcriptional regulators and membrane t...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2011.09.008
更新日期:2012-01-01 00:00:00
abstract::The major immunodominant integral outer membrane protein C (OmpC) from Salmonella typhi Ty21a was overexpressed, without the signal peptide, in Escherichia coli. The protein aggregates as inclusion bodies (IBs) in the cytoplasm. OmpC from IBs was solubilized with 4 M urea and refolded. This involved rapid dilution of ...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2004.12.023
更新日期:2005-03-01 00:00:00
abstract::Acyl carrier protein (ACP) was purified from Euglena gracilis variety bacillaris in yields of about 1 mg/100 g (wet wt) of cells. Antibodies against the purified protein were raised in hens and isolated from eggs. Antibodies raised against Euglena ACP inhibited the Euglena chloroplast nonaggregated fatty acid syntheta...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/1046-5928(91)90072-q
更新日期:1991-04-01 00:00:00
abstract::The membrane-anchored metalloproteinase ADAM17 (TNF-alpha converting enzyme; TACE; EC 3.4.24.86) continues to be an attractive drug target in inflammatory diseases and cancer. Cocrystallization of its catalytic domain with a lead compound was complicated by the tenacious retention of the prodomain that has been shown ...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2006.10.021
更新日期:2007-04-01 00:00:00
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
更新日期:2002-12-01 00:00:00