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 synthetic streptavidin gene, the installation of a transcription terminator, and the use of a sporulation mutant strain. In comparison with the wild-type streptavidin gene in expression studies, a combination of these approaches resulted in a 2.3-fold increase in streptavidin production. The production yields in complex and semidefined media were 94 and 24 mg/liter, respectively. A simple purification scheme which requires only a single ion-exchange matrix was designed to purify streptavidin to homogeneity directly from the culture supernatant. Purified streptavidin was in full length with good biotin binding capacity (3.2 binding sites available per tetramer). A combination of this expression system and purification scheme would be useful for production and purification of high-quality functional streptavidin for characterizations and practical applications.
journal_name
Protein Expr Purifjournal_title
Protein expression and purificationauthors
Wu SC,Hassan Qureshi M,Wong SLdoi
10.1006/prep.2001.1582keywords:
subject
Has Abstractpub_date
2002-04-01 00:00:00pages
348-56issue
3eissn
1046-5928issn
1096-0279pii
S1046592801915827journal_volume
24pub_type
杂志文章abstract::As an insect-selective neurotoxin, scorpion long-chain BjαIT is a promising prospect for insecticidal application; however, the difficulty of obtaining natural BjαIT represents the major obstacle preventing analysis of its insecticidal activity against agricultural insect pests. Here, we screened recombinant Pichia pa...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2018.07.013
更新日期:2018-12-01 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::Fab fragments isolated from papain digests of monoclonal antibodies have a wide variety of uses in analytical and in both in vivo and in vitro diagnostic applications. A novel, non-affinity method which uses the Gradiflow to purify Fab fragments from the papain digest of a mouse IgG1 anti-c-myc monoclonal antibody is ...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2003.07.005
更新日期:2003-12-01 00:00:00
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
更新日期:2003-01-01 00:00:00
abstract::Accurate diagnosis is essential for the treatment, prevention, and control of tuberculosis. Poor specificity of the tuberculin skin test in BCG-vaccinated populations and constraints to implementation of PCR and CMI-based diagnostic assays in developing countries warrant development of easy-to perform robust serologic...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2005.04.014
更新日期:2005-11-01 00:00:00
abstract::Chemokine receptors are a specific class of G-protein-coupled receptors (GPCRs) that control cell migration associated with routine immune surveillance, inflammation and development. In addition to their roles in normal physiology, these receptors and their ligands are involved in a large number of inflammatory diseas...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2009.03.001
更新日期:2009-07-01 00:00:00
abstract::Serine hydroxymethyltransferase (SHMT) plays a key role in cell physiology as it participates in the different interconversion pathway of folate coenzymes, provides almost exclusively folate one carbon fragments for the biosynthesis of a variety of end products. For the first time, Mycobacterium leprae glyA gene, enco...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2007.04.017
更新日期:2007-09-01 00:00:00
abstract::Escherichia coli glutamic acid decarboxylase is a pyridoxal phosphate-dependent enzyme that catalyzes the alpha-decarboxylation of glutamate to yield 4-amino-butyrate and CO2. The E. coli chromosome contains two genes encoding for this enzyme, gadA and gadB, which map at distinct loci. Their protein products differ in...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.1996.0121
更新日期:1996-12-01 00:00:00
abstract::This paper reports the overproduction and the details of a rapid method to purify active sigmaS monomers from a T7 RNA polymerase-based protein expression system. This 2-day procedure involves solubilizing inclusion bodies in sarkosyl detergent, removal of sarkosyl by dialysis, and a single gel filtration column chrom...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.1996.0069
更新日期:1996-08-01 00:00:00
abstract::The basidiomycete Phanerochaete chrysosporium produces several isoforms of lignin peroxidase, which catalyzes the oxidative depolymerization of lignin To date, ion-exchange chromatography and preparative isoelectric focusing (IEF) have been commonly used for isolation of lignin peroxidase isoenzymes. In this work we h...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.1995.1044
更新日期:1995-06-01 00:00:00
abstract::HSPC144 is a newly identified gene in human CD34(+) hematopoietic stem/progenitor cells. In this work, we have expressed and purified the 225-residue protein from Escherichia coli BL21 (DE3) and identified a stable fragment HSPC144-P (residues 44-225) by limited proteolysis method. The HSPC144-P fragment exhibits high...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2005.03.008
更新日期:2005-07-01 00:00:00
abstract::The covalent addition of 4-amino-4-deoxy-L-arabinose (L-Ara4N) groups to lipid A, which resides in the outer membranes of bacteria such as Salmonella typhimurium and Escherichia coli, is the final step in the polymyxin-resistance pathway in these organisms. This modification is catalyzed by the inner membrane protein ...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2005.08.028
更新日期:2006-03-01 00:00:00
abstract::Insoluble expression of intrinsically soluble proteins with native activity is potentially a promising alternative to soluble expression of folded protein or insoluble expression of unfolded protein requiring refolding. Here, we attempted to express highly soluble halophilic His-rich metal binding protein (HP) as inso...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2019.01.001
更新日期:2019-04-01 00:00:00
abstract::The availability of recombinant monomeric alkaline phosphatase (AP) is highly desirable in analytical applications involving AP fusion proteins. The cobalt-dependant alkaline phosphatase IV from Bacillus subtilis (BSAP), which was reported to be strongly monomeric, was overexpressed in Escherichia coli using pET autoi...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2013.06.008
更新日期:2013-08-01 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::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
更新日期:2012-08-01 00:00:00
abstract::Mammalian α-defensins contribute to innate immunity by exerting antimicrobial activity against various pathogens. To perform structural and functional analysis of α-defensins, large amounts of α-defensins are essential. Although many expression systems for the production of recombinant α-defensins have been developed,...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2015.04.007
更新日期:2015-08-01 00:00:00
abstract::Two DNA polymerase genes have been isolated from Thermococcus strains, Thermococcus zilligii from New Zealand, and the other, Thermococcus 'GT', a fast-growing strain isolated from the Galapagos trench. Both genes were isolated by genomic walking PCR, a technique that does not require expression of the gene product. P...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2006.07.022
更新日期:2007-03-01 00:00:00
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
更新日期:1994-06-01 00:00:00
abstract::Darbepoetin alfa is an engineered and hyperglycosylated analog of recombinant human erythropoietin (EPO) which is used as a drug in treating anemia in patients with chronic kidney failure and cancer. This study desribes the secretory expression of a codon-optimized recombinant form of darbepoetin alfa in Leishmania ta...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2015.10.013
更新日期:2016-02-01 00:00:00
abstract::The hexokinase (ATP;D-hexose 6-phosphotransferase, EC 2.7.1.1) of Schistosoma mansoni has been expressed in Escherichia coli as a fusion protein including an N-terminal polyhistidine tag and enterokinase cleavage site. The enzyme was purified by metal chelate chromatography to > 95% homogeneity, based on analysis by S...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.1996.0113
更新日期:1996-11-01 00:00:00
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
更新日期:2005-02-01 00:00:00
abstract::An expression library was generated from a partial NcoI and HindIII digest of genomic DNA from the thermophilic bacterium, Bacillus stearothermophilus P1. The DNA fragments were cloned into the expression vector pQE-60 and transformed into Escherichia coli M15[EP4]. Sequence analysis of a lipase gene showed an open re...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.2001.1456
更新日期:2001-08-01 00:00:00
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
更新日期:1993-02-01 00:00:00
abstract:INTRODUCTION:Glycodelin is a glycoprotein with different oligosaccharides that are responsible for its diverse biological functions in contraception and immunosuppression. Therefore, it is necessary to have access to adequate amounts of glycodelin with retained carbohydrate structure for functional studies because the ...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2016.09.020
更新日期:2017-02-01 00:00:00
abstract::A family of restriction enzyme- and ligation-independent cloning vectors has been developed for producing recombinant His-tagged fusion proteins in Escherichia coli. These are based on pURI2 and pURI3 expression vectors which have been previously used for the successful production of recombinant proteins at the millig...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2010.10.013
更新日期:2011-03-01 00:00:00
abstract::Human paraoxonase (hPON3) is a high density lipoprotein-related glycoprotein with multi-enzymatic properties and antioxidant activity which is proposed to participate in the prevention of low density lipoprotein (LDL) oxidation. In this study, hPON3 gene was amplified from Human Fetal Liver Marathon-Ready cDNA and exp...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2005.07.021
更新日期:2006-03-01 00:00:00
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
更新日期:2013-12-01 00:00:00
abstract::Keratinocyte growth factor (KGF) is a member of the fibroblast growth factor family that acts specifically on epithelial cells in a paracrine mode. We employed a mammalian expression system to synthesize recombinant human KGF and isolated two preparations, KGF-a and KGF-b, from medium conditioned by Chinese hamster ov...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.1997.0840
更新日期:1998-03-01 00:00:00
abstract::This is the first report of an insect esterase efficiently expressed in the methylotrophic yeast Pichia pastoris (so far insect esterases have been produced only in the baculovirus system). Having isolated a Tribolium castaneum carboxylesterase cDNA (TCE), we were initially unable to express it in Escherichia coli or ...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2005.04.011
更新日期:2005-08-01 00:00:00