| improvement of production, assay and purification of streptavidin. | the production of streptavidin by streptomyces avidinii in several different media was examined at 24, 48 and 72 hours. flask studies indicated that fermentation media containing either complex or multiple carbon sources resulted in higher yields of streptavidin than media with a single carbon source. streptavidin could be detected in crude fermentation broths by use of a tritiated biotin binding assay. this assay appears to give useful estimates of streptavidin production. depending upon the me ... | 1990 | 1366608 |
| streptavidin production by streptomyces avidinii in a laboratory fermenter. | | 1992 | 1633994 |
| production of streptavidin in a synthetic medium. | a simple, inexpensive procedure for producing streptavidin has been described. the biotin-binding protein was produced by growing streptomyces avidinii in a synthetic liquid culture medium containing l-asparagine as the sole nitrogen source. with this procedure, extraneous proteinaceous substances inherently present in culture media prepared with yeast extract or with peptones were not present to interfere with isolation and purification of streptavidin. when harvested after 7-8 days of incubati ... | 1988 | 2844909 |
| crystallographic data for streptomyces avidinii streptavidin. | crystallization conditions are reported for streptomyces avidinii streptavidin with and without bound biotin. x-ray examination of the free and bound crystal forms shows the streptavidin-biotin complex crystals to be most suitable for high resolution structure analysis. a complete x-ray data set to 2.6 a resolution was collected for the streptavidin-biotin crystals using a two-dimensional area detector. reduction and analysis of the x-ray diffraction pattern show that the complex crystallizes in ... | 1987 | 3624275 |
| an improved method for the single-step purification of streptavidin. | a new method for the preparation of a more efficient, stable iminobiotin-containing resin for the isolation of streptavidin was developed. cl-sepharose was activated with p-nitrophenyl chloroformate, and the resultant carbonate derivative was reacted with diaminohexane. subsequent reaction of the amino-containing resin with iminobiotin-n-hydroxysuccinimide ester (in an organic solvent) yielded the stable affinity resin. the capacity of this resin for either avidin or streptavidin was 12 mg per m ... | 1986 | 3772022 |
| immunoaffinity isolation of membrane antigens with biotinylated monoclonal antibodies and immobilized streptavidin matrices. | an efficient immunoaffinity method for isolating detergent-solubilized membrane antigens with murine and rat monoclonal antibodies is described that takes advantage of the high affinity (ka approximately 10(14) m-1) of streptavidin for biotin. streptavidin, a mr approximately 60,000 biotin-binding protein secreted by streptomyces avidinii, is covalently coupled to a high mr exclusion agarose gel (streptavigel) and used to adsorb soluble immune complexes formed with affinity-purified, biotinylate ... | 1984 | 6386987 |
| iminobiotin affinity columns and their application to retrieval of streptavidin. | a method is described for the retrieval of streptavidin from the culture broth of streptomyces avidinii. the key step in this procedure is the adsorption of streptavidin from culture concentrates to an affinity column in which iminobiotin is attached to ah-sepharose 4b. this column binds streptavbidin at ph 11 and releases the protein at ph 4. the recovery of streptavidin is practically quantitative. the ph dependence of the iminobiotin-avidin affinity, discovered by green [green, n. m. (1966) b ... | 1980 | 6933515 |
| structural relationship of streptavidin to the calycin protein superfamily. | streptavidin is a binding protein, from the bacteria streptomyces avidinii, with remarkable affinity for the vitamin biotin. the lipocalins and the fatty acid-binding proteins (fabps), are two other protein families which also act by binding small hydrophobic molecules. within a similar overall folding pattern (a beta-barrel with a repeated +1 topology), large parts of the lipocalin, fabp, and streptavidin molecules can be structurally equivalenced. the first structurally conserved region within ... | 1993 | 8224179 |
| secretion of streptavidin from bacillus subtilis. | streptavidin is an extracellular tetrameric protein produced by streptomyces avidinii. a series of hybrid gene fusions consisting of bacillus signal peptide coding regions fused to the mature streptavidin sequence were constructed. b. subtilis strains harboring these plasmids accumulate a tetrameric streptavidin in the growth medium. the properties of the streptavidin produced by b. subtilis are similar to those of the streptavidin produced by s. avidinii. b. subtilis strains carrying the variou ... | 1993 | 8285693 |
| construction and expression of a synthetic streptavidin-encoding gene in escherichia coli. | a synthetic gene encoding 'core' streptavidin (sav) [amino acid (aa) residues 13-140 of streptomyces avidinii sav] has been efficiently expressed in escherichia coli from the iptg-inducible lac promoter of plasmid pet3a. in this system, expression levels are nearly tenfold greater for the synthetic gene than for the corresponding native gene. the synthetic gene was constructed from overlapping oligodeoxyribonucleotides whose sequences were optimized to incorporate codons preferred by highly expr ... | 1993 | 8294009 |
| identification of biotinylated molecules using a baculovirus-expressed luciferase-streptavidin fusion protein. | a genetic fusion between streptavidin of streptomyces avidinii and luciferase of pyrophorus plagiophthalamus was constructed. the fusion protein was produced in the sf9 insect cell line using the baculovirus expression vector system (bevs). sodium dodecyl sulfate polyacrylamide gel electrophoresis of the proteins from cells infected with the recombinant virus, vl1393-lucgr-streptav, revealed that the fusion protein migrated with an apparent molecular weight of 75 kda. light emission measurements ... | 1996 | 8679206 |
| reduction of growth and acetyl-coa carboxylase activity by expression of a chimeric streptavidin gene in escherichia coli. | the streptavidin gene from streptomyces avidinii was expressed in e. coli as a non-fusion protein and as a glutathione s-transferase fusion protein. the streptavidin protein accumulated primarily in the inclusion bodies and did not alter cell growth. in contrast, the glutathione-s-transferase-streptavidin fusion protein was soluble. nondenaturing polyacrylamide gel electrophoresis indicated that the chimeric glutathione-s-transferase-streptavidin protein was present mostly as a monomer, with som ... | 1996 | 8867633 |
| a new approach for containment of microorganisms: dual control of streptavidin expression by antisense rna and the t7 transcription system. | the use of microorganisms in the open environment would be of less concern if they were endowed with programmed self-destruction mechanisms. here, we propose a new genetic design to increase the effectiveness of cell suicide systems. it ensures very tight control of the derepression of cell death by the combination of the bacteriophage t7 rna polymerase-lysozyme system and an inducible synthesis of antisense rna and the escherichia coli laci repressor. functionality of this regulatory concept wa ... | 1997 | 9037005 |
| [cloning of streptavidin gene from streptomyces avidinii and its expression in escherichia coli. secretion of streptavidin by e. coli cells]. | the streptavidin gene from streptomyces avidinii was cloned, an expression plasmid constructed, and a highly effective strain producer of streptavidin created. it was shown that the leader peptide of streptavidin ensures the effective secretion of this protein into the periplasmic space of escherichia coli cells. the degradation site of the leader peptide was detected. upon treatment with the total fraction of proteases secreted by s. avidinii into the culture medium, "core" streptavidin was obt ... | 1999 | 10382041 |
| phages from landscape libraries as substitute antibodies. | in 'landscape' phage, as in traditional phage-display constructs, foreign peptides or proteins are fused to coat proteins on the surface of a filamentous phage particle. unlike conventional constructs, however, each virion displays thousands of copies of the peptide in a repeating pattern, subtending a major fraction of the viral surface. the phage body serves as an interacting scaffold to constrain the peptide into a particular conformation, creating a defined organic surface structure ('landsc ... | 2000 | 10964989 |
| non-natural cell surface receptors: synthetic peptides capped with n-cholesterylglycine efficiently deliver proteins into mammalian cells. | protein toxins such as shiga toxin and cholera toxin penetrate into cells by binding small molecule-based cell surface receptors localized to cholesterol and sphingolipid-rich lipid raft subdomains of cellular plasma membranes. molecular recognition between these toxins and their receptors triggers endocytic protein uptake through endogenous membrane trafficking pathways. we report herein the synthesis of functionally related non-natural cell surface receptors comprising peptides capped with n-c ... | 2003 | 12526694 |
| dimer-tetramer transition between solution and crystalline states of streptavidin and avidin mutants. | the biotin-binding tetrameric proteins, streptavidin from streptomyces avidinii and chicken egg white avidin, are excellent models for the study of subunit-subunit interactions of a multimeric protein. efforts are thus being made to prepare mutated forms of streptavidin and avidin, which would form monomers or dimers, in order to examine their effect on quaternary structure and assembly. in the present communication, we compared the crystal structures of binding site w-->k mutations in streptavi ... | 2003 | 12837778 |
| antibiotic msd-235. i. production by streptomyces avidinii and streptomyces lavendulae. | | 1963 | 14274896 |
| affinity purification of streptavidin using tobacco mosaic virus particles as purification tags. | a new protein affinity purification system has been developed. recombinant tobacco mosaic virus (tmv) was used as an affinity matrix for isolation and purification of the given protein of interest. in model experiments, streptavidin-specific heptapeptide sequence tliahpq was inserted into tmv coat protein near the c end. this oligopeptide did not interfere significantly with viral replication, assembly, and movement. recombinant tmv functioned as an epitope tag recognizing streptavidin in plant ... | 2004 | 15450797 |
| an avidin-like domain that does not bind biotin is adopted for oligomerization by the extracellular mosaic protein fibropellin. | the protein avidin found in egg white seems optimized for binding the small vitamin biotin as a stable homotetramer. indeed, along with its streptavidin ortholog in the bacterium streptomyces avidinii, this protein shows the strongest known noncovalent bond of a protein with a small ligand. a third known member of the avidin family, as similar to avidin as is streptavidin, is found at the c-terminal ends of the multidomain fibropellin proteins found in sea urchin. the fibropellins form a layer k ... | 2005 | 15659374 |
| development of novel yeast cell surface display system for homo-oligomeric protein by coexpression of native and anchored subunits. | streptavidin derived from streptomyces avidinii was displayed on the cell surface of the yeast saccharomyces cerevisiae by cell-surface engineering using two types of plasmid for the expression of a native subunit and an anchored subunit fused with the c-terminus of 318 amino acids of flo1p containing a glycosylphosphatidylinositol anchor attachment signal. the displayed streptavidin had the binding ability for biotinylated compounds. this was confirmed by fluorescence microscopy after the adsor ... | 2006 | 16889375 |
| protein nanopatterns and biosensors using gold binding polypeptide as a fusion partner. | an efficient strategy for immobilizing proteins on a gold surface was developed by employing the gold binding polypeptide (gbp) as a fusion partner. using the enhanced green fluorescent protein (egfp), severe acute respiratory syndrome coronavirus (sars-cov) envelope protein (scvme), and core streptavidin (csa) of streptomyces avidinii as model proteins, specific immobilization of the gbp-fusion proteins onto the gold nanoparticles and generation of protein nanopatterns on the bare gold surface ... | 2006 | 17037921 |
| coupling of antibodies with biotin. | the avidin-biotin bond is the strongest known biological interaction between a ligand and a protein (kd = 1.3 x 10-15 m at ph 5.0) (1). the affinity is so high that the avidin-biotin complex is extremely resistant to any type of denaturing agent (2). biotin (see fig. 1) is a small, hydrophobic molecule that functions as a coenzyme of carboxylases (3). it is present in all living cells. avidin is a tetrameric glycoprotein of 66,000-68,000 molecular weight, found in egg albumin and in avian tissue ... | 2008 | 18287646 |
| an escherichia coli plasmid vector system for production of streptavidin fusion proteins: expression and bioselective adsorption of streptavidin-beta-galactosidase. | covalently immobilized biotin was used as a biospecific adsorbant to investigate the application of streptavidin as an affinity domain for simultaneous purification and immobilization of recombinant proteins. a streptavidin-beta-galactosidase fusion protein was constructed and tested as a model system. the gene for streptavidin from streptomyces avidinii was modified by polymerase chain reaction to mutate the stop codon and to facilitate cloning into an escherichia coli expression vector yieldin ... | 1994 | 18618647 |
| development of a whole-cell biosensor by cell surface display of a gold-binding polypeptide on the gold surface. | cell surface display was used as a strategy to display the gold-binding polypeptide (gbp) fusion protein on the surface of escherichia coli, and consequently to immobilize the cells on the gold surface. the dna encoding the gbp was fused to the truncated fadl gene and was expressed by the tac promoter. for the display of the core streptavidin (csa) of streptomyces avidinii, the csa gene was fused to the truncated oprf gene. after the dual display of fadl-gbp and oprf-csa on the surface of e. col ... | 2009 | 19228193 |
| characterization of cytotoxic compound from marine sediment derived actinomycete streptomyces avidinii strain su4. | to investigate the cytotoxic activity of actinomycete isolated from marine sediment. | 2012 | 23569845 |
| development of fed-batch strategies for the production of streptavidin by streptomyces avidinii based on power input and oxygen supply studies. | streptavidin is a tetrameric protein with an extremely high affinity to biotin and different biotin-like peptide-tags. this characteristic causes its widespread use in biotechnology. streptavidin is produced by the fermentation of wild type streptomyces avidinii or by recombinant streptomyces lavendulae, escherichia coli, and bacillus subtilis strains. however, little is known about the influence of power input and oxygen supply as well as feeding strategies on the production of streptavidin by ... | 2013 | 23142512 |
| [investigation of protein translocation sec-system with heterologous gene expression in shewanella oneidensis mr-1 bacterium cells]. | a comparison of the primary structures of the protein translocation sec-system proteins in the shewanella oneidensis mr-1 and escherichia coli bacteria was carried out. the process of translocation of recombinant pro-enteroxins (seb and seh) from staphylococcus aureus and pro-streptavidin (sav) from streptomyces avidinii in the s. oneidensis mr-1 and e. coli cell periplasm was studied. it was demonstrated that these marker proteins are transferred into the periplasmic space of the s. oneidensis ... | 2015 | 26204774 |
| secretory production of tetrameric native full-length streptavidin with thermostability using streptomyces lividans as a host. | streptavidin is a tetrameric protein derived from streptomyces avidinii, and has tight and specific biotin binding affinity. applications of the streptavidin-biotin system have been widely studied. streptavidin is generally produced using protein expression in escherichia coli. in the present study, the secretory production of streptavidin was carried out using streptomyces lividans as a host. | 2015 | 25582841 |
| the x-ray three-dimensional structure of avidin. | avidin is a basic, highly stable, homotetrameric protein, isolated from bird egg-white, binding up to four molecules of d-biotin with extremely high affinity (kd approximately 10(-15) m). the protein has been the object of different crystallographic investigations. in all the crystal structures, the four avidin subunits display almost exact 222 symmetry. each avidin chain (128 amino acids) is arranged in a eight-stranded antiparallel beta-barrel, whose inner region defines the d-biotin binding s ... | 1999 | 10796979 |
| genetic engineering of streptavidin, a versatile affinity tag. | streptavidin, a tetrameric protein produced by streptomyces avidinii, has been used as a useful, versatile affinity tag in a variety of biological applications. the efficacy of streptavidin is derived from its extremely high binding affinity for the vitamin biotin. for the last several years, we have used genetic engineering as a primary means to enhance the properties of streptavidin and to expand the application of streptavidin as an affinity tag. in this review, we describe several geneticall ... | 1998 | 9792500 |
| three-dimensional structure of the tetragonal crystal form of egg-white avidin in its functional complex with biotin at 2.7 a resolution. | the three-dimensional structure of hen egg-white avidin, crystallized in a tetragonal crystal form, has been solved at 2.7 a resolution by molecular replacement methods. after refinement the crystallographic r-factor is 16.8%, for the 7255 reflections in the 10.0 to 2.7 a resolution range. the asymmetric unit contains two avidin polypeptide chains (m(r) 2 x 15,600), which build up the functional tetramer through a crystallographic 2-fold axis parallel to the c unit cell direction. the avidin tet ... | 1993 | 8515446 |
| streptavidin blocks immune reactions mediated by fibronectin-vla-5 recognition through an arg-gly-asp mimicking site. | streptavidin is a biotin-binding analogue of egg-white avidin which is secreted by the bacterium streptomyces avidinii. we have recently reported that streptavidin contains an arg-tyr-asp-ser (ryds) sequence which exhibits structural homology to the arg-gly-asp-ser (rgds) cell adhesion domain of fibronectin and other matrix-associated glycoproteins. competition studies with rgd peptides indicated that streptavidin binds to cells via this site and that the binding is independent of biotin recogni ... | 1993 | 8096183 |
| using affinity capillary electrophoresis to determine binding stoichiometries of protein-ligand interactions. | we have developed a new method utilizing affinity capillary electrophoresis (ace) for the determination of binding stoichiometries in biochemical systems. using the same concentration of a ligand in the sample and the electrophoresis buffer, the appearance of an inverted peak corresponding to the free ligand in the resulting electropherogram provides a criterion of binding of a ligand to its receptor protein. for both low (fast off rates) and high (slow off rates) affinity systems, analysis of t ... | 1994 | 8075061 |
| [culture of streptomyces and purification of streptavidin]. | for preparation and purification of streptavidin, two synthetic mediums and a procedure for purifying streptavidin were studied. we found that streptomyces avidinii grew vigorously and showed typical colony characterization. at the sametime, these synthetic mediums were in favour of producing streptavidin with a maximal production of 15.2 micrograms/ml by using deae 52 column for purification of streptavidin. this purification procedure is very simple and easy, and could be widely used. | 1995 | 7657327 |
| molecular cloning and nucleotide sequence of the streptavidin gene. | using synthetic oligonucleotides as probes we have cloned the streptavidin gene from a genomic library of streptomyces avidinii. nucleotide sequence analysis indicated that a 2 kb dna-fragment contained the entire coding region, a signal peptide region and the 3' and 5' flanking regions of the gene. the deduced amino acid sequence shows several interrupted blocks of homology with the amino acid sequence of chicken egg-white avidin. analysis of the secondary structure suggests a high content of b ... | 1986 | 3951999 |
| isolation and characterization of highly purified streptavidin obtained in a two-step purification procedure from streptomyces avidinii grown in a synthetic medium. | a method is described for isolation of streptavidin from cultures of streptomyces avidinii grown in a synthetic culture medium for 6-10 days. streptavidin is precipitated directly from culture supernatant fluid using 80% ammonium sulfate, and the precipitate is dialyzed against water and centrifuged at 40,000 x g for 60 min. the absorbency coefficient at 280 nm of purified streptavidin was estimated to be 31.7142 +/- 0.1806 for a 1% solution. the protein appeared to be greater than 90% homogeneo ... | 1988 | 3049826 |
| postsecretory modifications of streptavidin. | streptavidin, an extracellular biotin-binding protein from streptomyces avidinii, exhibits a multiplicity in its electrophoretic mobility pattern which depends both upon the conditions for growth of the bacterium and upon the protocol used in the purification of the protein. the observed structural heterogeneity appears to reflect the action of two types of postsecretory molecular events: proteolytic digestion of the intact mr-18,000 subunit to a minimal molecular size (approx. mr 14,000), and a ... | 1989 | 2719654 |
| heterogeneity of ribosomal proteins among streptomyces species and its application to identification. | the ribosomal proteins from 11 streptomyces strains representing various numerical taxonomic clusters were compared by two-dimensional page. the protein patterns were specific for each species and were unaffected by acridine dye treatment, suggesting genetic stability of ribosomal proteins. an attempt was made to identify one strain of streptomyces by both traditional taxonomic methods and analysis of the ribosomal protein patterns. both methods identified the strain as streptomyces lavendulae, ... | 1989 | 2632667 |
| procedure for the purification of streptavidin by hydrophobic interaction chromatography. | a procedure is described for the purification of hydrophobic microbial proteins such as streptavidin from streptomyces avidinii, using benzyl-dc bead cellulose as the column material. the separation is rapid with a high loading capacity and sufficient resolution for preparative uses. advantages are discussed especially for industrial purposes. | 1990 | 2086584 |
| structural studies on avidinorubicin, a novel anthracycline with platelet aggregation inhibitory activity. | avidinorubicin (mw 1,214, c60h86n4o22) was isolated from the cultured broth of strain nr0576 (which was identified as streptomyces avidinii stapley et al.) by butyl alcohol extraction, sephadex lh-20 column chromatography and preparative hplc. avidinorubicin inhibited thrombin-induced platelet aggregation with an ic50 being 7.9 microm and was determined to be a novel anthracycline possessing two units of a new aminosugar, avidinosamine, in place of two decilonitrose groups in decilorubicin. | 1991 | 2071489 |
| expression of streptavidin in tomato resulted in abnormal plant development that could be restored by biotin application. | biotin is an essential cofactor for a variety of carboxylase and decarboxylase reactions and is involved in diverse metabolic pathways of all organisms. in the present study we tested the hypothesis that controlling biotin availability by the expression of streptomyces avidinii streptavidin, would impede plant development. transient expression of streptavidin fused to plant signal peptide, bacterial signal peptide or both, in tomato (lycopersicon esculentum cv. vf36) plants resulted in various l ... | 2004 | 15202718 |
| gold nanoparticles paper as a sers bio-diagnostic platform. | bioactive papers are usually challenged by four major limitations: sensitivity, selectivity, simplicity and strength (4s). gold nanoparticles (aunps) treated paper has previously been demonstrated as a surface enhanced raman scattering (sers) active substrate, capable of addressing the 4s issues. in this study, aunps on paper substrate were functionalized by a series of biomolecules to develop a generic sers platform for antibody-antigen detection. the functionalization steps were performed by t ... | 2013 | 23978290 |
| fed-batch production and secretion of streptavidin by hansenula polymorpha: evaluation of genetic factors and bioprocess development. | streptavidin - a protein secreted by the filamentous bacterium streptomyces avidinii - is applied in a variety of methods, leading to numerous studies on its heterologous production. development and characterization of a novel expression system for streptavidin genes by hansenula polymorpha is described utilizing different target gene variants along with the two methanol-inducible promoters pmox and pfmd. extracellular product concentrations were higher for cultivation at 30 instead of 37°c. the ... | 2016 | 26988393 |
| corrigendum to "development of fed-batch strategies for the production of streptavidin by streptomyces avidinii based on power input and oxygen supply studies" [j. biotechnol. 163 (2013) 325-332]. | | 2016 | 27080089 |
| molecular imaging and contrast agent database (micad) | white blood cells (wbcs) labeled with radioactive gallium (67ga), indium (111in), or technetium (99mtc) are often used for the detection of infections, but this application of wbcs is laborious and requires the use of autologous wbcs, which increases the probability of introducing an infection (1). in addition, this technique may not detect all types of infections. although computed tomography (ct) and magnetic resonance imaging (mri) are often used to characterize an infection or inflammation, ... | 2004 | 20641491 |
| molecular imaging and contrast agent database (micad) | biotin is a protein found in mammalian cells and is considered to be a vitamin because it has an important function in the cellular metabolism of lipids and carbohydrates. avidin is a protein that is found primarily in the egg white or in bacteria (known as streptavidin if isolated from bacteria; streptomyces avidinii is the source of this protein). although the two proteins have different origins, they are known to have a very strong non-covalent interaction (discovered almost seven decades ago ... | 2004 | 20641259 |
| avidin-biotin technology in targeted therapy. | the goal of drug targeting is to increase the concentration of the drug in the vicinity of the cells responsible for disease without affecting healthy cells. many approaches in cancer treatment are limited because of their broad range of unwanted side effects on healthy cells. targeting can reduce side effects and increase efficacy of drugs in the patient. | 2010 | 20233034 |
| cell-surface streptavidin fusion protein for rapid selection of transfected mammalian cells. | we developed a series of eight mammalian cell surface marker fusion genes by using the streptavidin gene from streptomyces avidinii. these fusion genes are useful and non-growth-toxic selection markers for rapid-harvest transfected mammalian cells. two streptavidin constructs were used; the longer fragment contains the native bacterial signal sequence, which the shorter fragment lacks. for expression of the streptavidin antigen on the surface of mammalian cells, streptavidin was flanked by a mam ... | 2007 | 17175122 |