| a new broad-spectrum aminoglycoside antibiotic complex, sporaricin. i. fermentation, isolation and characterization. | two new aminoglycoside antibiotics, sporaricins a (c17h35n5o5) and b (c15h32n4o4) produced by a rare actinomycetales, saccharopolyspora hirsuta subsp. kobensis have been isolated by column chromatography on a cation-exchange resin. sporaricin a is highly active against gram-positive and gram-negative bacteria including aminoglycoside-resistant strains. | 1979 | 37226 |
| a new broad-spectrum aminoglycoside antibiotic complex, sporaricin. ii. taxonomic studies on the sporaricin producing strain saccharopolyspora hirsuta subsp. kobensis nov. subsp. | morphological, cultural and physiological characteristics of a new nocardioform actinomycete are reported. the microorganism which produces the antibiotic complex sporaricin has been selectively isolated from a sample of soil obtained from kobe city, hyogo prefecture, japan. by whole-cell analysis of the actinomycete, meso-diaminopimelic acid, arabinose and galactose were identified. but lipid lcn-a (lipid characteristic of nocardia) and nocardomycolic acid were not detected. the taxonomic chara ... | 1979 | 457579 |
| a novel actinomycete from sugar-cane bagasse: saccharopolyspora hirsuta gen. et. sp. nov. | a new species of nocardioform actinomycete isolated from spontaneously heated sugar-cane bagasse is described as saccharopolyspora hirsuta gen. et sp. nov. it has affinities with species of both nocardia and actinomadura but can be distinguished from both genera by its morphology, sporulation, wall and lipid analyses, antibiotic resistance, degradation and carbon utilzation tests. | 1975 | 1097584 |
| effect of natural and synthetic immunomodulators on the synthesis of interferon by peritoneal cells of mice. | the effect of different natural and synthetic immunomodulators on the spontaneous interferon (ifn) synthesis by freshly isolated resident peritoneal cells of balb/c, nzb and c3h mice was investigated. actinomycetal glycolipids isolated from curtobacterium betae, faenia rectivirgula, rothia dentocariosa and saccharopolyspora hirsuta at the concentration 1-20 micrograms/ml were found to potentiate the ifn synthesis by the peritoneal cells of balb/c mice. similar results were obtained when dsrna, l ... | 1992 | 1279963 |
| promoter constructions for efficient secretion expression in streptomyces lividans. | promoters from different streptomyces genes were cloned in front of the tendamistat gene from s. tendae, in order to study secretion-expression in s. lividans using a pij702 plasmid vector system. besides the promoters we cloned a transcriptional terminator downstream of the tendamistat gene to improve transcription efficiency. the promoters we selected were: (1) a synthetic escherichia coli-like consensus promoter; (2) the aphi promoter of the neomycin resistance gene from s. fradiae; (3) an er ... | 1992 | 1368202 |
| targeted gene replacements in a streptomyces polyketide synthase gene cluster: role for the acyl carrier protein. | a methodology was developed to construct any desired chromosomal mutation in the gene cluster that encodes the actinorhodin polyketide synthase (pks) of streptomyces coelicolor a3(2). a positive selection marker (resistance gene) is first introduced by double crossing-over into the chromosomal site of interest by use of an unstable delivery plasmid. this marker is subsequently replaced by the desired mutant allele via a second high-frequency double recombination event. the technology has been us ... | 1992 | 1453961 |
| transforming growth factor-beta is generated in the course of hypersensitivity pneumonitis: contribution to collagen synthesis. | mice of the c57bl/6 strain were instilled with optimal doses (150 micrograms/day for 3 days/wk) of the thermophilic actinomycete faeni rectivirgula (also known as saccharopolyspora rectivirgula or micropolyspora faeni) to induce a hypersensitivity pneumonitis inflammation that mimics the human disease affecting certain occupational groups. this mouse model was characterized by a very significant alveolitis (3-fold increase in bronchoalveolar lavage [bal] cell number at 48 h and a 10-fold increas ... | 1992 | 1497904 |
| isolation and characterization of sporeamicin c. | | 1992 | 1500342 |
| cloning and analysis of a gene (sms13) encoding sannamycin b-glycyltransferase from streptomyces sannanensis and its distribution among actinomycetes. | a gene encoding sannamycin b-glycyltransferase (sms13) of streptomyces sannanensis ifo 14239 was identified by cloning and complementation of s. sannanensis mutant sn13 which is blocked at the interconversion of sannamycins b and a. the cloned dna fragment also permitted the conversion of fortimicin b to a both in s. sannanensis sn13 and streptomyces lividans tk23. dna sequences similar to sms13 were detected in all five producers of the fortimicin-group antibiotics, micromonospora olivasterospo ... | 1992 | 1517162 |
| prokaryotic calcium-binding protein of the calmodulin superfamily. calcium binding to a saccharopolyspora erythraea 20 kda protein. | the ef-hand calcium-binding protein from saccharopolyspora erythraea has been shown, using 113cd nmr, to possess three cd(2+)-ion binding sites. this indicates that of the four ef-hand motifs in the molecule, one (probably site 2) is unable to bind cd(2+)-ions. data from the titration of the protein with ca2+, in the presence of quin2, were fitted to a curve calculated on the assumption that the protein contains three high affinity ca2+ binding sites, two of which (pk1 = 8.0, pk2 = 9.0) are stro ... | 1992 | 1544472 |
| organization of the enzymatic domains in the multifunctional polyketide synthase involved in erythromycin formation in saccharopolyspora erythraea. | localization of the enzymatic domains in the three multifunctional polypeptides from saccharopolyspora erythraea involved in the formation of the polyketide portion of the macrolide antibiotic erythromycin was determined by computer-assisted analysis. comparison of the six synthase units (su) from the erya genes with each other and with mono- and multifunctional fatty acid and polyketide synthases established the extent of each beta-ketoacyl acyl-carrier protein (acp) synthase, acyltransferase, ... | 1992 | 1547954 |
| purification and characterization of tdp-d-glucose 4,6-dehydratase from anthracycline-producing streptomycetes. | tdp-d-glucose 4,6-dehydratase, which converts tdp-d-glucose to tdp-d-4-keto-6-deoxyglucose, was purified to near-homogeneity from the daunorubicin and baumycin-producing organism streptomyces sp. c5 (968-fold purification with a 41% recovery), and from the daunorubicin producer streptomyces peucetius atcc 29050 (1000-fold purification with a 37% recovery). the tdp-d-glucose 4,6-dehydratases from streptomyces sp. c5 and s. peucetius were determined by sds-page and hplc gel filtration to be homodi ... | 1992 | 1588310 |
| phage typing--a useful tool in actinomycete systematics. | a total of 905 strains of several genera of the actinomycetales were tested in respect to their sensitivity to genus- and species-specific actinophages. the results provided clear evidence that phage typing is a useful aid for the identification of actinomycetes. at the genus level, the specificity of actinophages has led to the correct placement of falsely identified strains, as well as to the rearrangement of genera. at the species level, however, phage typing has been less successful: only so ... | 1992 | 1612443 |
| biosynthesis of the erythromycin macrolactone and a rational approach for producing hybrid macrolides. | the three erya genes involved in the formation of the polyketide portion of the macrolide antibiotic erythromycin in saccharopolyspora erythraea, appear to be organized in a single transcriptional unit on the basis of the results of gene disruption experiments. an insertion sequence-like element of lower g + c content separates eryai from eryaii. the organization of the enzymatic domains present in the erya-encoded multifunctional polypeptides, determined by computer-assisted analysis, is presen ... | 1992 | 1612455 |
| identification of debs 1, debs 2 and debs 3, the multienzyme polypeptides of the erythromycin-producing polyketide synthase from saccharopolyspora erythraea. | the ery a region of the erythromycin biosynthetic gene cluster of saccharopolyspora erythraea has previously been shown to contain three large open reading frames (orfs) that encode the components of 6-deoxyerythronolide b synthase (debs). polyclonal antibodies were raised against recombinant proteins obtained by overexpression of 3' regions of the orf2 and orf3 genes. in western blotting experiments, each antiserum reacted strongly with a different high molecular weight protein in extracts of e ... | 1992 | 1618327 |
| sporeamicin a, a new macrolide antibiotic. i. taxonomy, fermentation, isolation and characterization. | sporeamicin a, a novel antibiotic, was isolated from the culture filtrate of an actinomycete. the producing organism, strain l53-18, was taxonomically assigned as a species of the genus saccaropolyspora. the antibiotic was extracted with chloroform and was then purified by crystallization. it was obtained as colorless prisms from ethanolic solutions. sporeamicin a exhibited a strong uv absorption peak at 276 nm. the molecular formula of sporeamicin a was determined to be c37h63no12. | 1992 | 1624360 |
| sporeamicin a, a new macrolide antibiotic. iii. biological properties. | sporeamicin a is a new erythromycin-type antibiotic isolated from a species of saccharopolyspora. it was active in vitro against a wide variety of gram-positive bacteria. in vitro studies indicated that the sporeamicin a was stable in the presence of human serum, although it was bound to serum proteins. sporeamicin a was effective in the mouse protection test against staphylococcus aureus, streptococcus pyogenes and streptococcus pneumoniae. sporeamicin a attained higher plasma and tissue levels ... | 1992 | 1624362 |
| isolation and characterization of sporeamicin b. | | 1992 | 1624386 |
| isolation and characterization of a sulfur-regulated gene encoding a periplasmically localized protein with sequence similarity to rhodanese. | during sulfur-limited growth, the cyanobacterium synechococcus sp. strain pcc 7942 loses most of its photosynthetic pigments and develops an increased capacity to acquire sulfate. sulfur deprivation also triggers the synthesis of several soluble polypeptides. we have isolated a prominent polypeptide of 33 kda that accumulates specifically under sulfur-limiting conditions. this polypeptide was localized to the periplasmic space. the gene for this protein (designated rhda) was isolated and discove ... | 1991 | 1708376 |
| regulation of secondary metabolism in streptomyces spp. and overproduction of daunorubicin in streptomyces peucetius. | two dna segments, dnrr1 and dnrr2, from the streptomyces peucetius atcc 29050 genome were identified by their ability to stimulate secondary metabolite production and resistance. when introduced into the wild-type atcc 29050 strain, the 2.0-kb dnrr1 segment caused a 10-fold overproduction of epsilon-rhodomycinone, a key intermediate of daunorubicin biosynthesis, whereas the 1.9-kb dnrr2 segment increased production of both epsilon-rhodomycinone and daunorubicin 10- and 2-fold, respectively. in a ... | 1992 | 1729206 |
| characterization of saccharopolyspora erythraea cytochrome p-450 genes and enzymes, including 6-deoxyerythronolide b hydroxylase. | previous studies of erythromycin biosynthesis have indicated that a cytochrome p-450 monooxygenase system is responsible for hydroxylation of 6-deoxyerythronolide b to erythronolide b as part of erythromycin biosynthesis in saccharopolyspora erythraea (a. shafiee and c. r. hutchinson, biochemistry 26:6204-6210 1987). the enzyme was previously purified to apparent homogeneity and found to have a catalytic turnover number of approximately 10(-3) min-1. more recently, disruption of a p-450-encoding ... | 1992 | 1732208 |
| 6-deoxyerythronolide-b synthase 2 from saccharopolyspora erythraea. cloning of the structural gene, sequence analysis and inferred domain structure of the multifunctional enzyme. | sequencing of the erya region of the erythromycin biosynthetic gene cluster from saccharopolyspora erythraea has revealed another structural gene (orf b), in addition to the previously characterised orf a, which appears to encode a component of 6-deoxyerythronolide-b synthase, the enzyme that catalyses the first stage in the biosynthesis of the polyketide antibiotic erythromycin a. the nucleotide sequence of orf b, which lies immediately adjacent to orf a, has been determined. the predicted gene ... | 1992 | 1740151 |
| biological characterization of induced phages from saccharopolyspora hirsuta 367 and comparison with phage jhj-1. | phages jhj-2 and jhj-3 were isolated from saccharopolyspora hirsuta 367 uc 8106 following induction with mitomycin c and amplified on s. hirsuta nrrl b-5792. their properties were compared with those of phage jhj-1, isolated previously from s. hirsuta 367 nrrl 12045. the dna restriction patterns appeared to be identical. one-step growth experiments showed no differences between the replication cycles. burst sizes ranged from 100 to 110 p.f.u. per cell. however, the three phages showed some diffe ... | 1991 | 1770351 |
| [effect of the methods of sterilization on the quality of nutrient media and on the level of antibiotic production]. | it was shown that conditions for heat sterilization of nutrient media for biosynthesis of antibiotics had an impact on their biochemical composition and activity of a fermentation broth in production of penicillin and erythromycin. the temperature of 120 degrees c and the time of 25 minutes proved to be optimal for sterilization of the media in regard to both preservation of their biochemical composition and providing of the maximum antibiotic productivity on the one hand and maintenance of the ... | 1991 | 1814272 |
| subcloning and expression of midecamycin polyketide synthase genes from streptomyces mycarofaciens 1748. | a 2.4 kb fragment containing the midecamycin polyketide synthase genes (mps) was subcloned from the preliminary clone pcn8b12 out of the genomic library of midecamycin-producing strain s. mycarofaciens 1748, by using the homologous dna of the actinorhodin polyketide synthase gene (act i) as hybridization probe. this dna fragment was subcloned onto streptomyces/e. coli shuttle vector pmhm3. a recombinant plasmid pcg2 was obtained. the transformation of the polyketide synthase deficient mutant of ... | 1991 | 1824237 |
| cloning of an aminoglycoside-resistance-encoding gene, kamc, from saccharopolyspora hirsuta: comparison with kamb from streptomyces tenebrarius. | an aminoglycoside-resistance-encoding gene (kamc) has been isolated from the sporaricin producer, saccharopolyspora (sac.) hirsuta, and expressed both in streptomyces lividans and escherichia coli. the pattern of resistance conferred by this gene was identical to that given by another gene (kamb) previously isolated from streptomyces tenebrarius. in accordance with the known action of the kamb product, the sac, hirsuta determinant also encodes a methyltransferase that modifies 16s rrna, thereby ... | 1991 | 1840536 |
| cloning and sequence analysis of genes involved in erythromycin biosynthesis in saccharopolyspora erythraea: sequence similarities between eryg and a family of s-adenosylmethionine-dependent methyltransferases. | the gene cluster (ery) responsible for production of the macrolide antibiotic erythromycin by saccharopolyspora erythraea is also known to contain erme, the gene conferring resistance to the antibiotic. the nucleotide sequence has been determined of a 4.5 kb portion of the biosynthetic gene cluster, from a region lying between 3.7 kb and 8.2 kb 3' of erme. this has revealed the presence of four complete open reading frames, including the previously known ery gene eryg, which catalyses the last s ... | 1991 | 1840640 |
| a new method for the rapid identification of genes encoding restriction and modification enzymes. | we have constructed derivatives of escherichia coli that can be used for the rapid identification of recombinant plasmids encoding dna restriction enzymes and methyltransferases. the induction of the dna-damage inducible sos response by the mcr and mrr systems, in the presence of methylated dna, is used to select plasmids encoding dna methyltransferases. the strains of e. coli that we have constructed are temperature-sensitive for the mcr and mrr systems and have been further modified to include ... | 1991 | 1851562 |
| [cytologic control of protoplast formation and regeneration in streptomyces erythraeus, strain btcc-2]. | experiments on protoplast formation and regeneration in s. erythraeus, strain btcc-2 (saccharopolyspora erythrae) were performed under microscopic control at all the stages. it was shown that the highest protoplast titer was provided by the mycelium grown in one step in the absence of glycine. for characterizing the protoplasts formed by the mycelium grown under different conditions, their regeneration capacity was estimated by microscopic examination of the protoplasts after 15-20-hour growth i ... | 1991 | 1877883 |
| production of h2o2 by alveolar macrophages in experimental allergic alveolitis. | experimental extrinsic allergic alveolitis (eaa) was induced in guinea pigs with saccharopolyspora rectivirgula. bronchoalveolar lavages were performed before inducing eaa (day 1, bal 1), on day 23 (bal 2), and on day 48 (bal 3). the number of cells/ml in lavage fluid was increased at bal 2 (4.79 x 10(6) and bal 3 (4.29 x 10(6)) compared with bal 1 (0.56 x 10(6)). the number of major cell types increased simultaneously, neutrophil becoming the predominant cell type over alveolar macrophages (am) ... | 1991 | 1886491 |
| tumor necrosis factor plays an essential role in determining hypersensitivity pneumonitis in a mouse model. | we examined the importance of the cytokine tumor necrosis factor-alpha (tnf-alpha) in a mouse model of hypersensitivity pneumonitis (hp). mice of the c57bl/6 strain were instilled intranasally 3 days/wk for 3 wk with 150 micrograms of the actinomycete faenia rectivirgula (micropolyspora faeni) to induce hp as a model of farmer's lung. this experimental model was associated with a progressive inflammation in the lungs of challenged mice, seen histologically as cellular infiltrates of large quanti ... | 1991 | 1931076 |
| an acyl-carrier-protein-thioesterase domain from the 6-deoxyerythronolide b synthase of saccharopolyspora erythraea. high-level production, purification and characterisation in escherichia coli. | the c-terminal region of a multifunctional polypeptide from the 6-deoxyerythronolide b synthase of saccharopolyspora erythraea is predicted to contain an acyl carrier protein and a thioesterase or acyltransferase activity [cortes, j., haydock, s. f., roberts, g. a., bevitt, d. j. & leadlay, p. f. (1990) nature 348, 176-178]. site-directed mutagenesis by means of the polymerase chain reaction was used to construct an efficient pt7-based expression plasmid for this domain. the recently developed t ... | 1991 | 1999200 |
| an erythromycin derivative produced by targeted gene disruption in saccharopolyspora erythraea. | derivatives of erythromycin with modifications at their c-6 position are generally sought for their increased stability at acid ph, which in turn may confer improved pharmacological properties. a recombinant mutant of the erythromycin-producing bacterium, saccharopolyspora erythraea, produced an erythromycin derivative, 6-deoxyerythromycin a, that could not be obtained readily by chemical synthesis. this product resulted from targeted disruption of the gene, designated eryf (systematic nomenclat ... | 1991 | 2011746 |
| modular organization of genes required for complex polyketide biosynthesis. | in saccharopolyspora erythraea, the genes that govern synthesis of the polyketide portion of the macrolide antibiotic erythromycin are organized in six repeated units that encode fatty acid synthase (fas)-like activities. each repeated unit is designated a module, and two modules are contained in a single open reading frame. a model for the synthesis of this complex polyketide is proposed, where each module encodes a functional synthase unit and each synthase unit participates specifically in on ... | 1991 | 2024119 |
| site-specific recombination in escherichia coli between the att sites of plasmid pse211 from saccharopolyspora erythraea. | pse211 from saccharopolyspora erythraea integrates site-specifically into the chromosome through conservative recombination between attp and attb, the plasmid and chromosomal attachment sites. integration depends on the presence of int, an open reading frame (orf) that lies adjacent to attp and encodes the putative integrase. immediately upstream of int lies xis (formerly called orf2) which encodes a basic protein that is thought to exhibit dna binding. xis and int were cloned in various combina ... | 1991 | 2046656 |
| cloning and characterization of the saccharopolyspora erythraea fdxa gene encoding ferredoxin. | the saccharopolyspora erythraea gene (fdxa) corresponding to a previously purified ferredoxin [shafiee and hutchinson, j. bacteriol., 170 (1988) 1548-1553] was cloned using an oligodeoxyribonucleotide probe based on the n-terminal sequence of the ferredoxin. the nucleotide sequence of a 1.3-kb segment encompassing fdxa indicates that the corresponding protein, sefdi, is 105 amino acids long, and very similar to other 7fe ferredoxins. a partial open reading frame closely linked to fdxa was also d ... | 1991 | 2055472 |
| cloning, characterization, and high-level expression in escherichia coli of the saccharopolyspora erythraea gene encoding an acyl carrier protein potentially involved in fatty acid biosynthesis. | the erythromycin a-producing polyketide synthase from the gram-positive bacterium saccharopolyspora erythraea (formerly streptomyces erythraeus) has evident structural similarity to fatty acid synthases, particularly to the multifunctional fatty acid synthases found in eukaryotic cells. fatty acid synthesis in s. erythraea has previously been proposed to involve a discrete acyl carrier protein (acp), as in most prokaryotic fatty acid synthases. we have cloned and sequenced the structural gene fo ... | 1991 | 2066335 |
| [cloning of clusters of erythromycin biosynthesis genes of streptomyces erythraeus (saccharopolyspora erythraea)]. | the erythromycin resistance gene (erme) and part of erythromycin biosynthesis genes located in the same cluster with the erme gene were cloned from s. erythraeus 3 subjected to improvement with respect to erythromycin production. for isolating the erythromycin biosynthesis genes, the plasmid vector puc18 and the phage vector lambda embl3 were used. the erme gene dna was used as a labeled probe for analysis of the recombinant plasmids and phages. the recombinant phages lambda erme1 and erme4 cont ... | 1990 | 2150305 |
| characterization of the genetic elements required for site-specific integration of plasmid pse211 in saccharopolyspora erythraea. | the 18.1-kilobase plasmid pse211 integrates into the chromosome of saccharopolyspora erythraea at a specific attb site. restriction analysis of the integrated plasmid, pse211int, and adjacent chromosomal sequences allowed identification of attp, the plasmid attachment site. nucleotide sequencing of attp, attb, attl, and attr revealed a 57-base-pair sequence common to all sites with no duplications of adjacent plasmid or chromosomal sequences in the integrated state, indicating that integration t ... | 1990 | 2180909 |
| organization of a cluster of erythromycin genes in saccharopolyspora erythraea. | we used a series of gene disruptions and gene replacements to mutagenically characterize 30 kilobases of dna in the erythromycin resistance gene (erme) region of the saccharopolyspora erythraea chromosome. five previously undiscovered loci involved in the biosynthesis of erythromycin were found, erybi, erybii, eryci, erycii, and eryh; and three known loci, eryai, eryg, and erme, were further characterized. the new ery phenotype, eryh, was marked by (i) the accumulation of the intermediate 6-deox ... | 1990 | 2185216 |
| mutation and cloning of eryg, the structural gene for erythromycin o-methyltransferase from saccharopolyspora erythraea, and expression of eryg in escherichia coli. | a mutant strain derived by chemical mutagenesis of saccharopolyspora erythraea (formerly known as streptomyces erythreus) was isolated that accumulated erythromycin c and, to a lesser extent, its precursor, erythromycin d, with little or no production of erythromycin a or erythromycin b (the 3"-o-methylation products of erythromycin c and d, respectively). this mutant lacked detectable erythromycin o-methyltransferase activity with erythromycin c, erythromycin d, or the analogs 2-norerythromycin ... | 1990 | 2185226 |
| cloning of genes involved in erythromycin biosynthesis from saccharopolyspora erythraea using a novel actinomycete-escherichia coli cosmid. | two plasmids were constructed that replicate in saccharopolyspora (sac.) erythraea, escherichia coli and streptomyces (s.) lividans, and used for the cloning of a locus involved in the synthesis of the macrolide antibiotic erythromycin (er). plasmid pal7002 contains the thiostrepton-resistance gene (tsr), a replicon-containing fragment from pjvi and puc9. plasmid pnji contains the lambda cos site but is otherwise similar to pal7002. a library of total dna from sac. erythraea was constructed in p ... | 1990 | 2199328 |
| phenotypic suppression by aminoglycoside antibiotics of mutations blocking erythromycin biosynthesis in saccharopolyspora erythraea. | the presence of a sublethal amount of apramycin, the aminoglycoside antibiotic used as a selectable marker in transformations of saccharopolyspora erythraea with pkc505 and related plasmids, was found to suppress phenotypically the s. erythraea eryb25 and eryb26 mutations blocking erythromycin biosynthesis in this organism, probably by the effect of mistranslation. | 1990 | 2228978 |
| an unusually large multifunctional polypeptide in the erythromycin-producing polyketide synthase of saccharopolyspora erythraea. | erythromycin a, a clinically important polyketide antibiotic, is produced by the gram-positive bacterium saccharopolyspora erythraea. in an arrangement that seems to be generally true of antibiotic biosynthetic genes in streptomyces and related bacteria like s. erythraea, the ery genes encoding the biosynthetic pathway to erythromycin are clustered around the gene (erme) that confers self-resistance on s. erythraea. the aglycone core of erythromycin a is derived from one propionyl-coa and six me ... | 1990 | 2234082 |
| disruption of a rhodaneselike gene results in cysteine auxotrophy in saccharopolyspora erythraea. | a 3,373-base-pair dna segment from a clone fortuitously isolated from saccharopolyspora erythraea by hybridization to an oligodeoxynucleotide probe was sequenced. computer-assisted analysis of the nucleotide sequence reveals three closely linked streptomyces open reading frames plus a fourth converging on the others. the deduced product of one of them, orf2, shows considerable similarity to bovine liver rhodanese. orf2, and the closely linked orf3 located just downstream of it, were disrupted by ... | 1990 | 2294090 |
| a repeated decapeptide motif in the c-terminal domain of the ribosomal rna methyltransferase from the erythromycin producer saccharopolyspora erythraea. | re-analysis of the primary structure of the ribosomal rna n-methyltransferase that confers self-resistance on the erythromycin-producing bacterium saccharopolyspora erythraea has confirmed the presence of a c-terminal domain containing extensive repeat sequences. nine tandem repeats can be discerned, with a decapeptide consensus sequence ggrx(h/r)gdrrt, although no single residue is wholly invariant. this highly polar, potentially flexible domain, which is predicted to adopt either a random coil ... | 1990 | 2335200 |
| saccharopolyspora hirsuta strain 367 releases jhj-1, a bacteriophage capable of propagation on old mycelium. | bacteriophage jhj-1 was isolated from saccharopolyspora hirsuta strain 367 nrrl 12045 as an endogenous but virulent phage. the plaque size was not self-limiting, since a few p.f.u. could completely lyse a lawn. electron microscopy showed that this phage belonged to group b of bradley's morphological classification. the jhj-1 genome is a linear dna molecule of 41.1 kbp with cohesive ends and a g + c content of 68.8-70.0 mol%. the dna cleavage map was established for 12 restriction endonucleases. ... | 1990 | 2391492 |
| macrolide antibiotic biosynthesis: isolation and properties of two forms of 6-deoxyerythronolide b hydroxylase from saccharopolyspora erythraea (streptomyces erythreus). | a cytochrome p-450 monooxygenase that catalyzes the hydroxylation of 6-deoxyerythronolide b, an intermediate of erythromycin a biosynthesis in saccharopolyspora erythraea (formerly streptomyces erythreus), was resolved into two forms, p-450i and p-450ii, by hydroxylapatite chromatography. these two proteins were purified to homogeneity from the ca 340 strain and found to have a p-450 content of 17.5 and 15.2 nmol/mg of protein, respectively. either enzyme catalyzed the nadph-dependent hydroxylat ... | 1987 | 2446657 |
| reverse transcriptase sequencing of 16s ribosomal rna from faenia rectivirgula, pseudonocardia thermophila and saccharopolyspora hirsuta, three wall type iv actinomycetes which lack mycolic acids. | the phylogenetic relationships of three mycolateless wall type iv actinomycetes, faenia rectivirgula, pseudonocardia thermophila and saccharopolyspora hirsuta, were examined using reverse transcriptase sequencing of 16s ribosomal rna. the sequences generated were aligned and the level of sequence homology calculated. the homology values were then use to produce a phylogenetic tree and to estimate s(ab) values for the construction of a dendrogram. both analyses show the three taxa to be closely r ... | 1988 | 2460582 |
| identification of a gene required for the terminal step in erythromycin a biosynthesis in saccharopolyspora erythraea (streptomyces erythreus). | we have identified a transcription unit in the erme region of the chromosome of the erythromycin (er)-producing bacterium saccharopolyspora erythraea (streptomyces erythreus) that is briefly switched on at about the time that macrolide production commences. disruption of the transcription unit, herein designated eryg, by insertion of an integrational plasmid vector, caused a block at the terminal step in the biosynthesis of erythromycin, the conversion of erythromycin c to a by o-methylation. | 1989 | 2469627 |
| coumamidines, new broad spectrum antibiotics of the cinodine type. i. discovery, taxonomy of the producing organism and fermentation. | coumamidines are water-soluble basic antibiotics related to the glycocinnamoylspermidines. they are produced by a soil isolate designated saccharopolyspora sp. ab 1167l-65. the coumamidines have broad spectrum activity and were selected in a screen for substances which inhibit pseudomonas aeruginosa. | 1989 | 2498266 |
| molecular characterization of a gene from saccharopolyspora erythraea (streptomyces erythraeus) which is involved in erythromycin biosynthesis. | a 7.3 kbp dna fragment, encompassing the erythromycin (em) resistance gene (erme) and a portion of the gene cluster encoding the biosynthetic genes for erythromycin biosynthesis in saccharopolyspora erythraea (formerly streptomyces erythraeus) has been cloned in streptomyces lividans using the plasmid vector pij702, and its nucleotide sequence has been determined using a modified dideoxy chain-termination procedure. in particular, we have examined the region immediately 5' of the resistance dete ... | 1989 | 2575703 |
| cloning, characterization, and heterologous expression of the saccharopolyspora erythraea (streptomyces erythraeus) gene encoding an ef-hand calcium-binding protein. | the regulatory effects of ca2+ in eucaryotic cells are mostly mediated by a superfamily of ca2+-binding proteins (cabs) that contain one or more characteristic ca2+-binding structural motifs, referred to as ef hands. we have cloned and sequenced the structural gene for an authentic ef-hand cab from the spore-forming gram-positive bacterium saccharopolyspora erythraea (formerly streptomyces erythraeus). when the gene was introduced into streptomyces lividans on the high-copy plasmid vector pij702 ... | 1989 | 2676983 |
| cloning of genes governing the deoxysugar portion of the erythromycin biosynthesis pathway in saccharopolyspora erythraea (streptomyces erythreus). | genes that govern the formation of deoxysugars or their attachment to erythronolide b and 3 alpha-mycarosyl erythronolide b, intermediates of the biosynthesis of the 14-membered macrolide antibiotic erythromycin, were cloned from saccharopolyspora erythraea (formerly streptomyces erythreus). segments of dna that complement the eryb25, eryb26, eryb46, eryc1-60, and eryd24 mutations blocking the formation of erythronolide b or 3 alpha-mycarosyl erythronolide b, when cloned in escherichia coli-stre ... | 1989 | 2681144 |
| hatomamicin (yl-0358m-a), a new alkaloid antibiotic: fermentation, isolation, structure, and biological properties. | hatomamicin, a new alkaloid antibiotic, was isolated from the culture filtrate of a strain of saccharopolyspora. the antibiotic was extracted with etoac and purified by silica gel column chromatography. the free alkaloid was obtained as pale yellowish prisms from ch3cn solution. it exhibits antimicrobial activity against gram-positive organisms. the apparent molecular formula of hatomamicin was determined to be c22h31no5. the structure has been established by a combination of spectroscopic and x ... | 1989 | 2753811 |
| methylation of 23s rrna caused by tlra (ermsf), a tylosin resistance determinant from streptomyces fradiae. | ribosomes from streptomyces griseofuscus expressing tlra, a resistance gene isolated from the tylosin producer streptomyces fradiae, are resistant to macrolide and lincosamide antibiotics in vitro. the tlra product was found to be a methylase that introduces two methyl groups into a single base within 23s rrna, generating n6,n6-dimethyladenine at position 2058. this activity is therefore similar to the erme resistance mechanism in saccharopolyspora erythraea (formerly streptomyces erythraeus). | 1989 | 2753855 |
| distribution of homologies among the genomes of several actinophages of faenia and saccharopolyspora as determined by dna hybridization. | the dna of phi fr114, a temperate phage of the thermophilic actinomycete faenia rectivirgula, was hybridized with the genomes of 11 different phages of faenia and saccharopolyspora. this revealed several regions (modules) within the phi fr114 dna which were distributed independently among the genomes of the other phages. the genome of the lytic phage 121, originally described for sap. erythraea, exhibited a similar modular organization. so far no functions of the possible modules are known. | 1989 | 2777538 |
| [studies on actinomycete flora and resources in the plateau lakes in yunnan]. | during the period of 1983 to 1987 mud and water samples were collected from 12 larger lakes on the yunnan plateau. the actinomycetes in both sediment and water samples were investigated. strains producing metabolites were screened by various procedures. the results are as follow: 1. the number and composition of actinomycetes in a lake were found to have relation to the physical and chemical features of the water-body. on the basis of the number and generic diversity of actinomycetes, lakes were ... | 1989 | 2800541 |
| site-specific integration in saccharopolyspora erythraea and multisite integration in streptomyces lividans of actinomycete plasmid pse101. | an 11.3-kilobase-pair plasmid, designated pse101, exists in saccharopolyspora erythraea nrrl 2338 as an integrated sequence (pse101int) at a unique chromosomal location and in the free form in less than an average of 1 copy per 10 chromosomes. the plasmid sequence is missing from s. erythraea nrrl 2359. restriction maps of the free and integrated forms of pse101 showed point-to-point correspondence. plasmid pect2 was constructed by ligation of pse101, pbr322, and the gene for thiostrepton resist ... | 1988 | 2834338 |
| bacteriophages of saccharopolyspora erythraea. | five bacteriophages infecting only saccharopolyspora erythraea (formerly streptomyces erythreus) among 43 streptomyces spp. tested were classified into two groups by phage-host relationships, restriction enzyme mapping, cohesive-end determinations, and southern hybridizations. phi se6, the most frequently isolated phage, produced clear plaques on all hosts tested, while phi se45, phi se57, phi se60, and phi se69 produced turbid plaques. phi se6 dna was linear, had a molecular weight of (27.6 +/- ... | 1987 | 3036768 |
| characterization of bacteriophage phi c69 of saccharopolyspora erythraea and demonstration of heterologous actinophage propagation by transfection of streptomyces and saccharopolyspora. | a bacteriophage, designated phi c69, isolated from a culture of saccharopolyspora erythraea was characterized. the phage propagates on sac. erythraea nrrl 2338 but does not infect 10 streptomyces or 3 micromonospora species tested. it infects sac. erythraea nrrl 2359 but does not produce infectious phage particles in this host. phi c69 is approximately 40 kb in length and contains cohesive ends. a cos fragment containing ligated phage dna ends was cloned in escherichia coli. restriction maps of ... | 1988 | 3073187 |
| purification and reconstitution of the electron transport components for 6-deoxyerythronolide b hydroxylase, a cytochrome p-450 enzyme of macrolide antibiotic (erythromycin) biosynthesis. | the hydroxylation of 6-deoxyerythronolide b (6d) to erythronolide b, a step in the biosynthesis of the 14-membered macrolide antibiotic erythromycin a by saccharopolyspora erythraea, is catalyzed by a cytochrome p-450 monooxygenase that requires two electron transport proteins for the function of this terminal hydroxylase (a. shafiee and c. r. hutchinson, biochemistry 26:6204-6210, 1987). two flavoproteins and an iron-sulfur protein (erythrodoxin) were purified from s. erythraea ca340 and shown ... | 1988 | 3127376 |
| purification of thymidine-diphospho-d-glucose 4,6-dehydratase from an erythromycin-producing strain of saccharopolyspora erythraea by high resolution liquid chromatography. | tdp-d-glucose 4,6-dehydratase was purified from saccharopolyspora erythraea, the producer of the macrolide antibiotic erythromycin a, by a high resolution chromatographic method that exploited the difference in the behavior of the protein on anionic exchange chromatography in tris/hcl or phosphate buffers. by this method, the enzyme was purified approximately 900-fold by two anionic exchange steps to more than 90% homogeneity. it was further purified to apparent homogeneity by hydrophobic intera ... | 1988 | 3170573 |
| the use of a chromosome integration vector to map erythromycin resistance and production genes in saccharopolyspora erythraea (streptomyces erythraeus). | the thiostrepton-resistance-conferring plasmid pij702 was integrated into the erme region of the chromosome of erythromycin (er)-producing bacterium saccharopolyspora erythraea (streptomyces erythraeus) by single, reciprocal (campbell) recombination between dna cloned in the vector and homologous nucleotide sequences in the chromosome. genetic mapping experiments by conjugational transfer were used to establish that the err gene, erme, was located close to the er-production loci erya34 and eryb2 ... | 1988 | 3220254 |
| characterization of se-3, a virulent bacteriophage of saccharopolyspora erythraea. | se-3 is a virulent bacteriophage isolated from a large-scale culture of saccharopolyspora erythraea, an erythromycin producer. the host range of the phage is narrow, limited to some strains of this species. another strain of sac. erythraea, and a strain of sac. hirsuta, are able to adsorb phage particles but do not sustain their complete multiplication. se-3 is closely related to the phage se-5 as shown by dna restriction mapping. the differences between se-3 and se-5 genomes are apparently limi ... | 1988 | 3246584 |
| transduction of plasmid dna in streptomyces spp. and related genera by bacteriophage fp43. | a segment (hft) of bacteriophage fp43 dna cloned into plasmid pij702 mediated high-frequency transduction of the resulting plasmid (prhb101) by fp43 in streptomyces griseofuscus. the transducing particles contained linear concatemers of plasmid dna. lysates of fp43 prepared on s. griseofuscus containing prhb101 also transduced many other streptomyces species, including several that restrict plaque formation by fp43 and at least two that produce restriction endonucleases that cut prhb101 dna. tra ... | 1988 | 3360743 |
| 2''-n-formimidoylsporaricin a produced by saccharopolyspora hirsuta subsp. kobensis. | | 1987 | 3558120 |
| menaquinone composition of mycolic acid-containing actinomycetes and some sporoactinomycetes. | the menaquinones of 141 actinomycetes representing the genera caseobacter, mycobacterium, nocardia, rhodococcus and some related taxa lacking mycolic acids were examined by mass spectrometry. the mycolic acid-containing strains were assigned to four groups on the basis of the predominant isoprenologue detected: rhodococcus coprophilus, r. equi, r. erythropolis, r. globerulus, r. rhodnii, r. rhodochrous and r. ruber contained dihydrogenated menaquinones with eight isoprene units; nocardia asteroi ... | 1985 | 3980298 |
| a new destomycin-family antibiotic produced by saccharopolyspora hirsuta. | | 1985 | 4008336 |
| ribosomal ribonucleic acid similarities in the classification of rhodococcus and related taxa. | duplexes were prepared between 14c-labelled rrna from both rhodococcus equi c7 and rhodococcus rhodochrous n54 and dna from 16 actinomycetes representing the genera rhodococcus, mycobacterium, nocardia, saccharopolyspora and streptomyces. the relationships between the organisms were determined by plotting the temperature at which 50% of the duplex was denatured (tm(e)) against the percentage of rrna binding (microgram 14c-labelled rrna duplexed per 100 micrograms filter-bound dna). all of the st ... | 1980 | 6160195 |
| saccharocin, a new aminoglycoside antibiotic. fermentation, isolation, characterization and structural study. | a new aminoglycoside antibiotic, saccharocin has been isolated from the fermentation broth of saccharopolyspora sp. ac-3440 (ferm p-6238) by column chromatography on a cation-exchange resin. saccharocin is active against gram-positive and gram-negative bacteria. the structure was elucidated to be 4"-deamino-4"-hydroxyapramycin by 13c nmr spectral analysis. | 1983 | 6874587 |
| vanoxonin, a new inhibitor of thymidylate synthetase. | vanoxonin, a new inhibitor of thymidylate synthetase, was found in cultured broths of the strain mg245-cf2 classified as saccharopolyspora hirsuta. vanoxonin, c18h25n3o9, was obtained as colorless powder. vanoxonin forms a vanadium complex which exhibits a strong inhibition against thymidylate synthetase. the concentration for 50% inhibition of the enzyme (ic50) was 0.7 micrograms/ml. | 1983 | 6874588 |
| a new broad-spectrum aminoglycoside antibiotic complex, sporaricin. iv. sporaricins c and d. | two new aminoglycoside antibiotics, sporaricins c and d have been isolated from the culture broth of saccharopolyspora hirsuta subsp. kobensis, which produced sporaricins a and b. the structures of sporaricins c and d have been determined to be 4-n-carbamoyl-glycylsporaricin b and 4-n-formylglycylsporaricin b, respectively. sporaricins c and d are active against gram-negative and gram-negative bacteria including aminoglycoside-resistant strains. | 1981 | 7287583 |
| the mrna for the 23s rrna methylase encoded by the erme gene of saccharopolyspora erythraea is translated in the absence of a conventional ribosome-binding site. | transcriptional analysis of the erme gene of saccharopolyspora erythraea, which confers resistance to erythromycin by n6-dimethylation of 23s rrna and which is expressed from two promoters, ermep1 and ermep2, revealed a complex regulatory region in which transcription is initiated in a divergent and overlapping manner. two promoters (eryc1p1 and eryc1p2) were identified for the divergently transcribed erythromycin biosynthetic gene eryc1, which plays a role in the formation of desosamine or its ... | 1994 | 7533884 |
| the use of pcr to isolate a putative abc transporter from saccharopolyspora erythraea. | a gene (ertx) encoding a putative abc transporter was cloned from the erythromycin producer saccharopolyspora erythraea, using pcr. the primers were based on regions of homology from abc transporters which confer resistance to macrolide antibiotics. while ertx encodes a protein with a strong degree of similarity to other macrolide abc transporters from streptomycetes and staphylococci, it did not confer resistance to erythromycin, tylosin, spiramycin, oleandomycin, josamcin, chalcomycin or midec ... | 1995 | 7557329 |
| in vivo analysis of straight-chain and branched-chain fatty acid biosynthesis in three actinomycetes. | the starter units for branched-chain and straight-chain fatty acid biosynthesis was investigated in vivo in three actinomycetes using stable isotopes. branched-chain fatty acids, which constitute the majority of the fatty acid pool, were confirmed to be biosynthesized using the amino acid degradation products methylbutyrl-coa and isobutyrl-coa as starter units. straight-chain fatty acids were shown to be constructed using butyrl-coa as a starter unit. isomerization of the valine catabolite isobu ... | 1995 | 7557334 |
| a novel insertion sequence (is)-like element of the thermophilic bacterium ps3 promotes expression of the alanine carrier protein-encoding gene. | a novel insertion sequence (is)-like element was found in the 5'-upstream region of the alanine carrier protein-encoding gene (acp) in the thermophilic bacterium ps3 chromosomal dna. the sequence contained an open reading frame (orf) encoding a polypeptide of 369 amino acids which revealed high similarity with orfs from is891 from the cyanobacterium anabaena and is1136 from saccharopolyspora erythraea. the direction of transcription was the same as that of acp, and typical inverted and direct re ... | 1995 | 7557457 |
| thermophilic actinomycetes in cane sugar mills: an aeromicrobiologic and seroepidemiologic study. | aerial prevalence of clinically important thermophilic actinomycetes and occurrence of precipitating antibodies against them in sera of 153 exposed workers have been reported. the study was carried out in two cane sugar mills namely, the upper doab sugar mills and the ramala sugar mills, located in north-west india. in both the sugar mills, t. sacchari was the predominant species, it accounted for 55.1% and 50.3% of the total population of thermophilic actinomycetes, followed by t. vulgaris (19. ... | 1995 | 7574549 |
| pc:ps liposomes induce a recruitment of neutrophils and the release of tnf alpha in the lungs of mice sensitized with saccharopolyspora rectivirgula. | the aim of this study was to verify the effect of nasally instilled liposomes (l) or dexamethasone-containing l (ldexa) on normal or inflamed lung tissue. three groups of mice were studied. group i was given saline instillations for 3 weeks prior to the instillation with liposomes. in groups ii and iii lung inflammation was induced by repeated instillations of saccharopolyspora rectivirgula before the instillation of liposomes (group ii) or liposomes containing dexamethasone (group iii). animals ... | 1995 | 7628521 |
| erythromycin biosynthesis. highly efficient incorporation of polyketide chain elongation intermediates into 6-deoxyerythronolide b in an engineered streptomyces host. | feeding of (2s,3r)-[2,3-13c2]-2-methyl-3-hydroxypentanoyl nac thioester (1a) to the recombinant organism streptomyces coelicolor ch999/pck7 harboring the complete set of erya genes from saccharopolyspora erythraea encoding the 6-deoxyerythronolide b synthase (debs) resulted in the formation of 6-deoxyerythronolide b (2a) labeled with 13c at c-12 and c-13, as evidenced by the appearance of a pair of enhanced and coupled doublets in the 13c nmr spectrum. the level of 13c enrichment was 15-20 atom% ... | 1995 | 7649863 |
| a cytochrome p450-like gene possibly involved in oleandomycin biosynthesis by streptomyces antibioticus. | a cosmid clone from an oleandomycin producer, streptomyces antibioticus, contains a large open reading frame encoding a type i polyketide synthase subunit and an oleandomycin resistance gene (oleb). sequencing of a 1.4-kb dna fragment adjacent to oleb revealed the existence of an open reading frame (olep) encoding a protein similar to several cytochrome p450 monooxygenases from different sources, including the products of the eryf and eryk genes from saccharopolyspora erythraea that participate ... | 1995 | 7737473 |
| cloning and heterologous expression of the genes encoding nonspecific electron transport components for a cytochrome p450 system of saccharopolyspora erythraea involved in erythromycin production. | the fora gene encoding a protein that can function as a nadh:ferredoxin oxidoreductase (for) has been cloned from saccharopolyspora erythraea, the erythromycin a (era) producer. in a previous study for protein, together with the fdxa ferredoxin from the same organism, was shown to be able to reconstitute the cytochrome p450 system responsible for the hydroxylation of 6-deoxyerythronolide b, an intermediate of era biosynthesis. nucleotide sequence data suggest that the cloned fora gene codes for ... | 1995 | 7737500 |
| structure of cytochrome p450eryf involved in erythromycin biosynthesis. | cytochrome p450eryf catalyzes the 6s-hydroxylation of 6-deoxyerythronolide b, the initial reaction in a multistep pathway to convert 6-deoxyerythronolide b into the antibiotic, erythromycin. the overall structure of p450eryf is similar to that of p450cam but differs in the exact positioning of several alpha-helices. the largest difference occurs in the b' helix and results in the enlargement of the substrate-binding pocket of p450eryf. the substrate is positioned with the macrolide ring perpendi ... | 1995 | 7749919 |
| estimation of the kinetic constants and elucidation of trends in growth and erythromycin production in batch and continuous cultures of saccharopolyspora erythraea using curve-fitting techniques. | the kinetics of erythromycin production were dependent on the identity of the growth rate-limiting nutrient during batch cultures of saccharopolyspora erythraea. semilogarithmic linear regression provided a single estimate of growth rate during the exponential phase, but partial cubic spline curve fit derivatives provided time-dependent specific growth and production rate profile. non-growth-linked product formation was observed when the medium was glucose- or phosphate-limited. however, growth- ... | 1993 | 7763960 |
| repositioning of a domain in a modular polyketide synthase to promote specific chain cleavage. | macrocyclic polyketides exhibit an impressive range of medically useful activities, and there is great interest in manipulating the genes that govern their synthesis. the 6-deoxyerythronolide b synthase (debs) of saccharopolyspora erythraea, which synthesizes the aglycone core of the antibiotic erythromycin a, has been modified by repositioning of a chain-terminating cyclase domain to the carboxyl-terminus of debs1, the multienzyme that catalyzes the first two rounds of polyketide chain extensio ... | 1995 | 7770773 |
| preliminary crystallographic analysis of an enzyme involved in erythromycin biosynthesis: cytochrome p450eryf. | cytochrome p450eryf was overexpressed in escherichia coli and purified in high yield. crystals of the protein in the presence of the substrate, 6-deoxyerythronolide b, have been obtained by the hanging drop vapor diffusion method, using polyethylene glycol 4000 as a precipitant. the crystals belong to the orthorhombic space group p2(1)2(1)2(1) with unit cell dimensions of a = 54.16 a, b = 79.67 a, and c = 99.48 a and one molecule per asymmetric unit. a complete native data set has been collected ... | 1994 | 7846029 |
| overproduction and characterization of the erythromycin c-12 hydroxylase, eryk. | hydroxylation of c-12 is one of the final steps in the biosynthesis of erythromycin a (era). a point of uncertainty in the erythromycin pathway has been whether the c-12 hydroxylase operates on each of two possible substrates, erythromycin b (erb) and erythromycin d (erd). stassi et al. have cloned the gene, designated eryk, which encodes the p-450 monooxygenase responsible for erythromycin c-12 hydroxylation in saccharopolyspora erythraea [stassi, d., donadio, s., staver, m. j., & katz, l. (199 ... | 1995 | 7849045 |
| amino acid sequence analysis of ribosomal protein at-l30 from members of the family pseudonocardiaceae. | the phylogenetic relationships of the genera belonging to the family pseudonocardiaceae were examined by a novel approach, amino acid sequencing of ribosomal at-l30 proteins. the results of partial amino acid sequencing of at-l30 preparations revealed that the members of the family pseudonocardiaceae are divided into four clusters; the first cluster contains the genus actinopolyspora, the second cluster contains the genus saccharopolyspora, the third cluster contains the genus amycolatopsis, and ... | 1995 | 7857791 |
| cloning of the genes encoding thymidine diphosphoglucose 4,6-dehydratase and thymidine diphospho-4-keto-6-deoxyglucose 3,5-epimerase from the erythromycin-producing saccharopolyspora erythraea. | genes involved in deoxysugar metabolism, encoding thymidine diphospho (tdp)-glucose 4,6-dehydratase (gdh) and a putative tdp-4-keto-6-deoxyglucose 3,5-epimerase (kde), were cloned from the erythromycin (er)-producing saccharopolyspora erythraea by means of an oligodeoxynucleotide corresponding to a segment of the purified gdh protein. determination of the nucleotide sequence established that kde lies 3' to gdh. the function of gdh was confirmed by an enzymatic assay following expression of the g ... | 1995 | 7883182 |
| biosynthesis of erythromycin: origin of the methyl protons. | 13c-nuclear magnetic resonance (13c-nmr) spectroscopy has been used to locate deuterium atoms incorporated biosynthetically into erythromycin from [3-13cd3]- and [3-13c]sodium propionate and l-[13cd3]- and l-[13c]methionine in saccharopolyspora erythraea jcm 4748. the alpha-shifts of deuterated methyl groups were clearly observed in broad-band deuterium and proton-decoupled 13c-nmr spectra. the seven propionate-derived methyl groups and the four methionine-derived methyl groups were shown to und ... | 1994 | 7923474 |
| dna sequence and functions of the actvi region of the actinorhodin biosynthetic gene cluster of streptomyces coelicolor a3(2). | six open reading frames (orfs) were identified by dna sequencing of 5.7 kilobase pairs at the left end of the act cluster (the so-called "actvi region"), in the order: orfb, orfa, orf1, orf2, orf3, orf4. orf1-4 are transcribed rightward and in the same direction as the orfs of the actva region which lies to the right of the actvi region, whereas orfa and orfb run in the opposite direction. by complementation of mutants and gene disruption of the wild type strain, the two previously genetically c ... | 1994 | 7929165 |
| domain v of 23s rrna contains all the structural elements necessary for recognition by the erme methyltransferase. | the erme methyltransferase from the erythromycin-producing actinomycete saccharopolyspora erythraea dimethylates the n-6 position of adenine 2058 in domain v of 23s rrna. this modification confers resistance to erythromycin and to other macrolide, lincosamide, and streptogramin b antibiotics. we investigated what structural elements in 23s rrna are required for specific recognition by the erme methyltransferase. the erme gene was cloned into r1 plasmid derivatives, providing a means of inducible ... | 1994 | 7961464 |
| analysis of five tylosin biosynthetic genes from the tyllba region of the streptomyces fradiae genome. | the tyllba region of the tylosin biosynthetic gene cluster of streptomyces fradiae contains at least five open reading frames (orfs). orf1 (tyli) encodes a cytochrome p450 and mutations in this gene affect macrolide ring hydroxylation. the product of orf2 (tylb) belongs to a widespread family of proteins whose functions are speculative, although tylb mutants are defective in the biosynthesis or addition of mycaminose during tylosin production. orfs 3 and 4 (tyla1 and tyla2) encode delta tdp-gluc ... | 1994 | 7984112 |
| egta induces the synthesis in escherichia coli of three proteins that cross-react with calmodulin antibodies. | escherichia coli mutants, (vera, dila) specifically resistant to the ca2+ channel inhibitors verapamil and diltiazem, respectively, are hypersensitive to egta and bapta. we have shown, using 1-d and 2-d gel electrophoresis, that the synthesis of at least 25 polypeptides in the mutants was enhanced by treatment with ca2+ chelators and the synthesis of at least 11 polypeptides was repressed. this pattern of induction was not observed in heat- or sds-treated cells and therefore does not appear to b ... | 1994 | 7997161 |
| distribution of oxoacyl synthase homology sequences within streptomyces dna. | a genomic dna sequence of streptomyces strain isp 5485 was cloned, sequenced and compared with corresponding information from nuclei acid data banks. the dna sequence was unique, but showed homology to dna coding for the condensing enzyme, 2-oxoacyl synthase, of the deoxyerythronolide b synthase complex (debs) from saccharopolyspora erythraea nrrl 2338. a subfragment of the sequenced dna was used to construct a gene-specific probe that formed part of the putative 2-oxoacyl synthase gene. the pcr ... | 1994 | 8020754 |
| occurrence of thermophilic actinomycetes in natural substrates in nigeria. | thermophilic actinomycetes were isolated from 163 (48.95%) of 333 samples of vegetable substrates and soil from different sites in anambra and enugu states in nigeria. thermoactinomyces (tha.) vulgaris was the most common, occurring in 32.4% of samples while tha. thalpophilus was isolated from 20.1%. tha. sacchari, saccharomonospora (sam.) viridis and saccharopolyspora (sap.) rectivirgula were isolated from 3-10.5% of the samples examined. streptomyces (stm.) thermovulgaris occurred in 2.7% and ... | 1994 | 8060118 |
| [isolation and characterization of genetic recombinants in protoplast fusion in saccharopolyspora erythraea]. | formation of genetic recombinants after conjugation and protoplast fusion in saccharopolyspora erythraea, an organism producing erythromycin, was studied comparatively. after the protoplast fusion the frequency of all the classes of the haploid recombinants increased 10 to 460 times by comparison with the conjugation. the protoplast fusion was characterized by higher diversity of the recombinant classes, up to 45.5 per cent of the recombinants being formed at the account of multiple crossing ove ... | 1993 | 8060179 |
| conjugal transfer of cosmid dna from escherichia coli to saccharopolyspora spinosa: effects of chromosomal insertions on macrolide a83543 production. | cosmid poj436, containing large inserts of saccharopolyspora spinosa (ss) dna, was transferred by conjugation from escherichia coli to ss an integrated into the chromosome, apparently by homologous recombination, at high frequencies (10(-5) to 10(-4) per recipient). transfer was mediated by the plasmid rp4 (rk2) transfer functions in e. coli, and the rk2 orit function located on poj436 [bierman et al., gene 116 (1992) 43-49]. poj436 lacking ss dna, or containing a small insert (approx. 2 kb) of ... | 1994 | 8063103 |
| divalent metal ion requirements of a thermostable multimetal beta-galactosidase from saccharopolyspora rectivirgula. | to understand the roles of metal ions on the catalytic properties and thermostability of the thermostable beta-galactosidase of saccharopolyspora rectivirgula, a thermophilic actinomycete, we have investigated the binding kinetics and requirements of divalent metal ions by equilibrium dialysis, titration, and metal ion buffer techniques. we found that the monomeric multimetal enzyme (m(r) 136,977) had eight specific binding sites for divalent metal ions. these sites were classified as follows: a ... | 1994 | 8071323 |
| characterisation of a streptomyces antibioticus gene encoding a type i polyketide synthase which has an unusual coding sequence. | a gene (orfb) from streptomyces antibioticus (an oleandomycin producer) encoding a large, multifunctional polyketide synthase (pks) was cloned and sequenced. its product shows an internal duplication and a close similarity to the third subunit of the pks involved in erythromycin biosynthesis by saccharopolyspora erythraea, showing the equivalent nine active site domains in the same order along the polypeptide. an unusual feature of this orf is the gc content of most of the sequence, which is sur ... | 1994 | 8107683 |