Abstract:
:Production of His-BbST [hexahistidine-tagged BbST (bubaline somatotropin)] by auto-induction in high-density shake-flask cultures coupled with a single-step, on-column purification and refolding strategy is described here. To optimize expression of BbST, different media and expression conditions were tested. The highest expression levels of BbST, exceeding 30% of the total Escherichia coli cellular proteins, were achieved when YNG and M9NG media were used. Using these auto-inducing media, the final concentration of BbST increased up to 455 mg/l and was severalfold higher than that obtained by isopropyl beta-D-thiogalactoside induction. Most of the target protein, however, was in the form of inclusion bodies, which were solubilized in 8 M urea solution (pH 8.5). Using immobilized-metal-ion-affinity chromatography, His-BbST could be purified from solubilized sample to >97% homogeneity in a single step in a biologically active state as judged by its efficient growth-promoting activity in Nb2 rat lymphoma cell proliferation assays. The expression and purification scheme, presented here, has a potential of scaling up to obtain pure and biologically active His-BbST relatively inexpensively for further studies and applications.
journal_name
Biotechnol Appl Biochemjournal_title
Biotechnology and applied biochemistryauthors
Sadaf S,Khan MA,Akhtar MWdoi
10.1042/BA20060154subject
Has Abstractpub_date
2007-05-01 00:00:00pages
21-6issue
Pt 1eissn
0885-4513issn
1470-8744pii
BA20060154journal_volume
47pub_type
杂志文章abstract::Sephadex was derivatized consecutively with Staphylococcus Protein A (SpA) and cell-specific antibodies, and the binding of cells to the resulting material was examined. For comparison, cell binding to commercially obtained SpA-Sepharose was determined. Sephadex G-10, carboxylated by reaction with glycine and activate...
journal_title:Biotechnology and applied biochemistry
pub_type: 杂志文章
doi:10.1042/ba20010062
更新日期:2002-02-01 00:00:00
abstract::Two different enzymes that metabolize lactate in the presence of oxygen, either to acetate plus CO2 (lactate 2-mono-oxygenase; Lmox) or to pyruvate plus H2O2 (lactate oxidase; Lox) were encapsulated in human and murine red blood cells (RBCs). Lmox shows a low affinity for lactate (Km 22 mM) and thus works at a low rat...
journal_title:Biotechnology and applied biochemistry
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doi:
更新日期:1995-12-01 00:00:00
abstract::A sensitive label-free electrochemical aptasensor was successfully fabricated for thrombin detection with platinum nanoparticles (Pt) and blocking reagent horseradish peroxidase (HRP) as enhancers. A Nafion®-graphene-coated electrode was first modified with an electrochemical probe of methylene blue (MB) through elect...
journal_title:Biotechnology and applied biochemistry
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doi:10.1002/bab.33
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abstract::The purpose of the present study was to investigate the effects of garlic (Allium sativum L.) on the diabetic nephropathy and oxidative stress induced by STZ (streptozotocin) in rats. Diabetes was induced in Male Sprague-Dawley rats by administering a single intraperitoneal injection of STZ (60 mg/kg of body weight). ...
journal_title:Biotechnology and applied biochemistry
pub_type: 杂志文章
doi:10.1042/BA20080086
更新日期:2009-03-01 00:00:00
abstract::The pH optimum, temperature-dependence, thermal stability, substrate specificity and subsite affinities of the 66 kDa, pI 4.0 glucoamylase of the filamentous fungus Trichoderma reesei were determined. It had a pH optimum of 5.5 and a temperature optimum (5 min reaction time) of 70 degrees C with soluble starch as subs...
journal_title:Biotechnology and applied biochemistry
pub_type: 杂志文章
doi:10.1111/j.1470-8744.1995.tb00333.x
更新日期:1995-04-01 00:00:00
abstract::Solid-supported bilayer lipid membranes (s-BLMs) that possess some properties similar to those of conventional BLMs can be self-assembled on a freshly cleaved metal wire by a two-step procedure: (i) The tip of a Teflon-coated platinum wire, while immersed in a lipid solution, is cut off with a scalpel; (ii) the new ti...
journal_title:Biotechnology and applied biochemistry
pub_type: 杂志文章,评审
doi:
更新日期:1990-10-01 00:00:00
abstract::A sensitive sandwich enzyme immunoassay (e.i.a.) for serrapeptase (TSP), an orally available anti-inflammatory proteinase, was established using affinity-purified anti-TSP rabbit IgG and its Fab' fragment conjugated with horseradish peroxidase as the first and the second antibodies respectively. TSP in the plasma was ...
journal_title:Biotechnology and applied biochemistry
pub_type: 杂志文章
doi:
更新日期:1994-08-01 00:00:00
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journal_title:Biotechnology and applied biochemistry
pub_type: 杂志文章
doi:10.1002/bab.1083
更新日期:2013-01-01 00:00:00
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journal_title:Biotechnology and applied biochemistry
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doi:
更新日期:1987-04-01 00:00:00
abstract::Yeast cytochrome c peroxidase (CcP) was purified from baker's yeast and immobilised onto a nylon membrane. The kinetics of the soluble and immobilised forms of the enzyme were investigated for the catalysed oxidation of potassium ferrocyanide in the presence of H2O2 and m-chloroperoxybenzoic acid. The pH dependence of...
journal_title:Biotechnology and applied biochemistry
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doi:
更新日期:1991-08-01 00:00:00
abstract::Parkinson's disease (PD) is the most common neurodegenerative movement disorder and is characterized by the loss of dopaminergic neurons and the formation of eosinophilic intracytoplasmic inclusion bodies known as Lewy bodies. Although alpha-synuclein is known to be a pivotal factor implicated in the pathogenesis of P...
journal_title:Biotechnology and applied biochemistry
pub_type: 杂志文章
doi:10.1042/ba20020004
更新日期:2002-08-01 00:00:00
abstract::The gene (lacS) encoding a thermostable beta-galactosidase enzyme (Ss beta-gal) from the archaebacterium Sulfolobus solfataricus has been cloned and expressed in simian CV1 and murine NIH3T3 cell lines. The recombinant protein is an active enzyme that shows the same properties of thermophilicity and thermostability as...
journal_title:Biotechnology and applied biochemistry
pub_type: 杂志文章
doi:
更新日期:1994-04-01 00:00:00
abstract::Empty fruit bunch (EFB) of oil palm trees was converted to fermentable sugars by the combined use of dilute acids and whole fungal cell culture-catalyzed hydrolyses. EFB (5%, w/v) was hydrolyzed in the presence of 0.5% H2 SO4 and 0.2% H3 PO4 at 160 °C for 10 Min. The solid fraction was separated from the acid hydrolys...
journal_title:Biotechnology and applied biochemistry
pub_type: 杂志文章
doi:10.1002/bab.1188
更新日期:2014-07-01 00:00:00
abstract::Single base mismatched oligonucleotides related to the rpoB gene of Mycobacterium tuberculosis, the mutations of which cause drug resistance of the infectious agent, were detected and discriminated using a surface plasmon resonance biosensor system. Thiol-modified oligonucleotides of the selected sequence (the probe) ...
journal_title:Biotechnology and applied biochemistry
pub_type: 杂志文章
doi:10.1002/bab.1101
更新日期:2013-07-01 00:00:00
abstract::A protein with a molecular mass of 66 kDa was isolated by a simple, rapid, and inexpensive method, using 3-N-morpholinopropanesulfonic acid, potassium thiocyanate, and dithiothreitol, from a mixture of spores, parasporal crystals, and cell debris of Bacillus thuringiensis subsp. kurstaki. The protein was active agains...
journal_title:Biotechnology and applied biochemistry
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doi:
更新日期:1990-06-01 00:00:00
abstract::Bacillus subtilis strain DM-03, which is isolated from starter culture used for the production of alcohol by local Assam tribes, grows optimally at 52-55 degrees C and secretes a significant amount of alpha-amylase at pH 8.0 into the culture media. This alpha-amylase, purified by ion-exchange, gel-filtration and rever...
journal_title:Biotechnology and applied biochemistry
pub_type: 杂志文章
doi:10.1042/BA20040034
更新日期:2004-12-01 00:00:00
abstract::Rapid progress has been made in the design and synthesis of oligomers and polymers that emulate the properties of natural proteins. Molecular bioengineering offers the chance to design and produce artificial polymeric proteins with tailored polymeric properties. The elastin-like polypeptides are a well-defined family ...
journal_title:Biotechnology and applied biochemistry
pub_type: 杂志文章
doi:10.1042/BA20050114
更新日期:2005-12-01 00:00:00
abstract::HIV stimulates strong immune CD8(+) cytotoxic T lymphocytes (CTL) response in infected people, despite causing an immunodeficiency. It has been demonstrated that this response could be very important for the control of the virus. We have shown previously that a recombinant fowlpox virus (rFWPV), expressing the multi-e...
journal_title:Biotechnology and applied biochemistry
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doi:10.1042/ba20020027
更新日期:2002-12-01 00:00:00
abstract::Three peptides complementary to exposed regions of two low-M(r) phosphotyrosine protein phosphatase isoenzymes and of the acylphosphatase muscle isoenzyme have been synthesized. Each peptide was synthesized on two different types of resins; the peptides were anchored to the resins by amide linkages. The peptide resins...
journal_title:Biotechnology and applied biochemistry
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doi:
更新日期:1994-10-01 00:00:00
abstract::Mirabilis himalaica is an endangered medicinal plant species in the Tibetan Plateau. The two genes respectively encoding chalcone synthase (MhCHS) and chalcone isomerase (MhCHI) were isolated and characterized from M. himalaica. The sequence analysis revealed that the two genes were similar with their corresponding ho...
journal_title:Biotechnology and applied biochemistry
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journal_title:Biotechnology and applied biochemistry
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doi:
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journal_title:Biotechnology and applied biochemistry
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doi:
更新日期:1986-04-01 00:00:00
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journal_title:Biotechnology and applied biochemistry
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journal_title:Biotechnology and applied biochemistry
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更新日期:2007-05-01 00:00:00
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journal_title:Biotechnology and applied biochemistry
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journal_title:Biotechnology and applied biochemistry
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更新日期:2004-08-01 00:00:00
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journal_title:Biotechnology and applied biochemistry
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更新日期:2014-03-01 00:00:00
abstract::Mechano-growth factor (MGF), an alternative splicing variant of insulin-like growth factor-1 (IGF-1) gene, promotes cell proliferation and inhibits cell differentiation. It also plays an important role in tumor development. It is important to optimize the production process and achieve MGF protein because there is no ...
journal_title:Biotechnology and applied biochemistry
pub_type: 杂志文章
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更新日期:2015-09-01 00:00:00
abstract::G-protein-coupled receptors (GPCRs), the largest family of cell-surface molecules involved in a number of biological and pathological processes, have recently emerged as key players in carcinogenesis and cancer progression. Orphan G protein-coupled receptors (oGPCRs) are a group of proteins lacking endogenous ligands....
journal_title:Biotechnology and applied biochemistry
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更新日期:2015-11-01 00:00:00