Abstract：
:The superoxide dismutases (EC 1.15.1.1) are a family of enzymes that catalyze the dismutation of superoxide radical anion to dioxygen and hydrogen peroxide. The active site contains a critical metal ion such as manganese, iron, or copper. The copper-containing protein also has one zinc ion bound per subunit. The standard method used to remove the metal ions from Cu,Zn superoxide dismutase has been to exhaustively dialyze the protein against chelating agents at low pH. We have developed a new method where the protein is bound to ion-exchange medium based on iminodiacetic acid immobilized on Sepharose. The bound protein is treated with a buffer containing edta at pH 3.5 to remove metal ions; the buffer is then exchanged for acetate buffer to remove edta, after which the protein is eluted by a salt gradient. An advantage of this method is that a single chromatography step is sufficient to produce apo protein. Results are shown for both human and bovine dimeric Cu,Zn superoxide dismutase and the monomeric Escherichia coli Cu,Zn superoxide dismutase. In every case, the metals were removed efficiently.
journal_name
Protein Expr Purifjournal_title
Protein expression and purificationauthors
Sutter B,Bounds PL,Koppenol WHdoi
10.1006/prep.2000.1241keywords:
subject
Has Abstractpub_date
2000-06-01 00:00:00pages
53-6issue
1eissn
1046-5928issn
1096-0279pii
S1046-5928(00)91241-5journal_volume
19pub_type
杂志文章abstract：:Single-stranded DNA-binding proteins (SSBs) play essential roles in DNA replication, recombination, and repair in bacteria, archaea, and eukarya. We report here the identification, expression, and purification of the SSB-like proteins of the thermophilic bacteria Thermus thermophilus and T. aquaticus. The nucleotide (...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/s1046-5928(02)00504-1
更新日期：2002-10-01 00:00:00
abstract：:The catalase gene of Psychrobacter sp. T-3 was cloned, and the gene product (PktA) was overexpressed in Escherichia coli. The specific activity of the purified PktA was slightly lower than that of the native purified enzyme obtained from Psychrobacter sp. T-3. Spectrophotometric measurements of the purified enzymes su...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2008.03.016
更新日期：2008-06-01 00:00:00
abstract：:Acetyl-CoA C-acetyltransferase synthase gene (AACT) cDNA, DNA and promoter were cloned from Sanghuangporus baumii. The gene ORF (1260 bp) encoded 419 amino acids. The AACT DNA includes five exons (1-84 bp, 140-513 bp, 570-1027 bp, 1090-1282 bp, 1344-1494 bp) and four introns (85-139 bp, 514-569 bp, 1028-1089 bp, 1283-...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2020.105592
更新日期：2020-06-01 00:00:00
abstract：:Pullulanase is crucial to the specific hydrolysis of branch points in amylopectin and is generally employed as an important enzyme in the starch-processing industry. Recombinant Bacillus subtilis that employs an inducible promoter would be a suitable candidate for pullulanase expression because of its safety and contr...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2018.03.012
更新日期：2018-08-01 00:00:00
abstract：:The metalloprotease PrtV from Vibrio cholerae serves an important function for the bacteria's ability to invade the mammalian host cell. The protein belongs to the family of M6 proteases, with a characteristic zinc ion in the catalytic active site. PrtV constitutes a 918 amino acids (102 kDa) multidomain pre-pro-prote...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2014.01.012
更新日期：2014-04-01 00:00:00
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
更新日期：2013-11-01 00:00:00
abstract：:Co-expression offers an important strategy for producing multiprotein complexes for biochemical and biophysical studies. We have found that co-expression of histones H2A and H2B (from yeast, chicken or Drosophila) leads to production of soluble heterodimeric H2AH2B complexes. Drosophila histones H3 and H4 can also be ...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2010.03.013
更新日期：2010-08-01 00:00:00
abstract：:Whey acidic proteins (WAP) belong to a large gene family of antibacterial peptides that perform critical immune system functions. The function of human epididymis protein 4 (HE4), a 124-amino acid long polypeptide that has two whey acidic protein four-disulfide core (WFDC) domains, is not well studied. Here, a fusion ...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2014.08.004
更新日期：2014-10-01 00:00:00
abstract：:The role of the termini of protein sequences is often perturbed by remnant amino acids after the specific protease cleavage of the affinity tags and/or by the amino acids encoded by the plasmid at/around the restriction enzyme sites used to insert the genes. Here we describe a method for affinity purification of a met...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2019.03.009
更新日期：2019-07-01 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：:Endostatin, a 20kDa C-terminal fragment of collagen XVIII, is a potent anti-angiogenic protein and inhibitor of tumor growth. Recombinant endostatin was prepared from Escherichia coli deposited as insoluble, inactive inclusion bodies. In the present study, we produced soluble and biologically active recombinant human ...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2004.09.021
更新日期：2005-06-01 00:00:00
abstract：:The type I antifreeze proteins are simple amphipathic helical proteins found in abundance in polar fish species, where they act to prevent freezing of internal fluids by a mechanism of noncolligative freezing point depression. Large-scale production of these proteins for research and biotechnological purposes has been...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1006/prep.1999.1040
更新日期：1999-06-01 00:00:00
abstract：:The erm proteins confer resistance to the MLS (macrolide-lincosamide-streptogramin B) antibiotics in various microorganisms, including pathogens, through dimethylation of a single adenine residue (A2085: Bacillus subtilis coordinate) of the 23S rRNA to reduce the affinity of antibiotics, thereby enabling the cells to ...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1006/prep.2002.1621
更新日期：2002-06-01 00:00:00
abstract：:The high specific lysyl endopeptidase (Lys-C; EC 3.4.21.50) is often used for the initial fragmentation of polypeptide chains during protein sequence analysis. However, due to its specificity it could be a useful tool for the production of tailor-made protein hydrolysates with for example bioactive or techno functiona...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2015.09.024
更新日期：2016-02-01 00:00:00
abstract：:Vibriolysin, an extracellular protease of Vibrio proteolyticus, is synthesized as a preproenzyme. The N-terminal propeptide functions as an intramolecular chaperone and an inhibitor of the mature enzyme. Extracellular production of recombinant vibriolysin has been achieved in Bacillus subtilis, but not in Escherichia ...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2008.08.001
更新日期：2008-12-01 00:00:00
abstract：:The purpose of this work was to characterize the functions of two sesquiterpene synthase genes from moso bamboo (Phyllostachys edulis). Two novel sesquiterpene synthase genes, belonging to the Tpsa subfamily, were isolated from moso bamboo. MoTPS2 was 1641 bp in length and encoded a protein of 63 kDa, whereas MoTPS6 w...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2015.11.019
更新日期：2016-04-01 00:00:00
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
更新日期：2008-10-01 00:00:00
abstract：:Campylobacteriosis is a disease in humans caused by the infection from Campylobacter spp. Human cases are mainly due to Campylobacter jejuni, although C. coli can cause gastroenteritis in humans as well. The bacteria are commensal in chicken tract and can be contaminated into chicken products during processing. Obviou...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2021.105818
更新日期：2021-04-01 00:00:00
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
更新日期：2004-06-01 00:00:00
abstract：:Resistance to Inhibitors of Cholinesterase-8 (Ric-8) proteins are molecular chaperones that fold heterotrimeric G protein α subunits shortly after biosynthesis. Ric-8 proteins also act as test tube guanine nucleotide exchange factors (GEF) that promote Gα subunit GDP for GTP exchange. The GEF and chaperoning activitie...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2018.10.002
更新日期：2019-02-01 00:00:00
abstract：:In this work, we present the production of an active 43 aa recombinant human beta-defensin-1 (rhBD-1(43)) in Escherichia coli AD202 cells using specific pLMM1-rhBD-1 expression system. Unique solubility properties of the C-terminal fragment of light meromyosin (LMM) allowed us to overcome foreseeable problems with iso...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2004.04.024
更新日期：2004-09-01 00:00:00
abstract：:A highly efficient Escherichia coli expression system was established to obtain an appreciable quantity of antihypertensive peptide. The DNA-coding sequence for the Gly-Val-Tyr-Pro-His-Lys peptide was chemically synthesized and linked to form a ten-copy in tandem. It was cloned into the vector pET-15b and expressed in...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2015.05.001
更新日期：2015-09-01 00:00:00
abstract：:The expression of human lactoferrin in the mammary gland is an attractive approach to diminish its current production cost. Previous attempts to produce lactorferrin in the milk of transgenic animals resulted in very high cost and uncertain results. In this paper, we have directly infused replication-defective adenovi...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2006.11.019
更新日期：2007-05-01 00:00:00
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
更新日期：2015-11-01 00:00:00
abstract：:Organophosphorus hydrolase (OPH) is a ∼38kDa enzyme encoded by opd gene of Flavobacterium sp. The enzyme can hydrolyze and inactivate variety of organophosphate (OP)-compounds, including chemical warfare nerve agents. Thus, OPH is a strong candidate for the development of therapeutic intervention against OP-poisoning ...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2015.01.012
更新日期：2015-07-01 00:00:00
abstract：:Protein phosphatase 2A (PP2A) contains a 36-kDa catalytic subunit (PP2Ac), a 65-kDa structural subunit (PR65/A), and a regulatory B subunit. The core enzyme consists of the structural and catalytic subunits. The catalytic subunit exists as two closely related isoforms, alpha and beta. Several natural toxins, including...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2005.06.002
更新日期：2006-01-01 00:00:00
abstract：:A growing number of studies require the purification of multiple proteins simultaneously and the development of simple economical high-throughput purification methods is essential. We have tested the purification of two related proteins in a variety of conditions to benchmark the semi-automated affinity chromatography...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2014.01.014
更新日期：2014-04-01 00:00:00
abstract：:The gene encoding a novel glucoamylase (GlucaM) from the Corallococcus sp. strain EGB was cloned and heterologous expressed in Escherichia coli BL21(DE3), and the enzymatic characterization of recombinant GlucaM (rGlucaM) was determined in the study. The glucaM had an open reading frame of 1938 bp encoding GlucaM of 6...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2015.06.009
更新日期：2017-01-01 00:00:00
abstract：:Novel GSH-AP (phenoxyl agarose coated gigaporous polystyrene, Agap-co-PSt) microspheres were successfully prepared by introducing GSH ligand into hydrophilic AP microspheres pre-activated with 1,4-butanediol diglycidyl ether. The gigaporous structure and chromatographic properties of GSH-AP medium were evaluated and c...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2013.11.004
更新日期：2014-03-01 00:00:00
abstract：:Disulfide bonds are normally formed after a polypeptide has been exported from the reducing environment of the cytoplasm into a more oxidizing compartment, such as the bacterial periplasm. Recently, we showed that in Escherichia coli trxB gor mutants, in which the reduction of thioredoxin and glutathione is impaired, ...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1006/prep.2001.1520
更新日期：2001-11-01 00:00:00
