Marine actinomycete Streptomyces sp. ISP2-49E,a new source of Rhamnolipid

Marine derived actinomycetes were isolated from sediment samples of Galveston Bay, Texas and screened for production of bioactive metabolites. Streptomyces sp. ISP2-49E, identified by 16s rDNA, produced a previously described Rha-Rha-C10-C10 rhamnolipid; however, this is the first report of a rhamnolipid produced by a Streptomyces sp. This organism was isolated from hydrocarbon contaminated sediment and further supports the trend of rhamnolipid production as a potential adaptation to this environment.     

IM-2, a butyrolactone autoregulator,induces production of several nucleoside antibiotics in Streptomyces sp. FRI-5

IM-2 is one of the butyrolactone autoregulators of Streptomyces, which triggers production of a blue pigment in Streptomyces sp. FRI-5 at a concentration of 0.6 ng/ml. In the absence of IM-2, Streptomyces sp. FRI-5 was found to produce d-cycloserine. However, the addition of IM-2 at 5-h cultivation stopped both growth and d-cycloserine production, and instead induced production of several different antibiotics. The IM-2-induced antibiotics were isolated from the culture broth, and assigned as the nucleoside antibiotics, showdomycin and minimycin. Therefore, IM-2 was concluded to be a global regulator of a secondary metabolism, which not only induced the production of nucleoside antibiotics but also suppressed d-cycloserine production.     

PI factor, a novel type quorum-sensing inducer elicits pimaricin production in Streptomyces natalensis

A chemically novel autoinducer (PI factor) has been purified from cultures of the pimaricin producer Streptomyces natalensis ATCC27448. The chemical structure of the PI molecule was identified as 2,3-diamino-2,3-bis (hydroxymethyl)-1,4-butanediol. Pimaricin biosynthesis in S. natalensis npi287, a mutant impaired in pimaricin production, was restored by supplementation with either A-factor from Streptomyces griseus IFO13350 or with PI factor. S. natalensis did not synthesize A-factor. The PI autoinducer was active at very low concentrations (50-350 nm). A threshold level of 50 nm was required to observe the induction effect. The dose-response curve was typical of a quorum-sensing type mechanism. The biosynthesis of PI factor was associated with cell growth of S. natalensis, both in defined and complex media. Supplementation of the wild-type S. natalensis with pure PI (300 nm) resulted in a stimulation of 33% of the production of pimaricin. These results indicate that the endogenous synthesis of PI factor is limiting for pimaricin biosynthesis in the wild-type strain. This water-soluble PI factor belongs to a novel class of autoinducers in Streptomyces species different from the classical butyrolactone autoinducers. Because restoration of pimaricin production in the npi287 mutant is conferred by both A-factor and PI factor, S. natalensis appears to be able to integrate different quorum signals from actinomycetes.     

Ansaetherone, a new radical scavenger from Streptomyces sp

We identified a new radical scavenger, ansaetherone (C26 H33 NO7), from a culture of the Streptomyces sp. USF-4727 strain. In our previous study, it was shown that this strain produced four lipoxygenase inhibitors, tetrapetalones A, B, C and D. The chemical structure of ansaetherone was elucidated by the spectroscopic method, indicating that this compound was constructed with an aglycon and a sugar moiety. This chemical structure suggested that ansaetherone was related to the tetrapetalones. This finding provided information regarding tetrapetalone biosynthesis. Ansaetherone showed radical scavenging activity with an ED50 value of 300 microM in our assay.     

Cloning of a DNA fragment involved in pigment production in Streptomyces avermitilis

Streptomyces avermitilis has the ability to synthesize a diffusible, brown, melanin-like pigment, a common property among many Streptomyces species. A region of the S. avermitilis chromosome involved in the production of this pigment was cloned in Escherichia coli. Production of the brown pigment was attained in E. coli, and is optimal when medium is supplemented with copper ions, tyrosine and IPTG. The cloned S. avermitilis pigment-producing DNA fragment is under the control of the lac promoter carried in the E. coli vector. The gene involved in pigment production could be used as a tool to analyse gene expression in S. avermitilis, and as an alternative cloning marker in Streptomyces-Escherichia coli vectors.     

Formation of new antibiotic, virenomycin, by a culture of Streptomyces virens sp. nev

A culture of a new species Streptomyces virens was isolated from a soil sample. It produced an antibiotic designated as virenomycin. The antibiotic was mainly synthesized in the mycelium. Only insignificant amounts of it were found in the culture fluid. The optimal nutrient medium for production of virenomycin contained glycerol, soybean meal, ammonium sulphate, sodium chloride and calcium carbonate. Crystalline virenomycin had a comparatively low antitumor activity and narrow spectrum.     

Streptomyces sp. Z-11-6--a new producer of extracellular L-glutamate oxidase

Glutamate oxidase activity was studied in 1254 Streptomyces strains isolated from the zonal soils of various regions of Russia and other countries. Seven strains proved to be producers of extracellular L-glutamate oxidase. The most active producer strain was identified, and the conditions of enzyme biosynthesis were optimized. A multistep-mutagenesis and selection procedure allowed a genetically stable strain, Streptomyces sp. Z-11-6, to be obtained, whose glutamate oxidase activity was 40 times higher than that of the original natural isolate.     

abaA, a new pleiotropic regulatory locus for antibiotic production in Streptomyces coelicolor.

Production of the blue-pigmented antibiotic actinorhodin is greatly enhanced in Streptomyces lividans and Streptomyces coelicolor by transformation with a 2.7-kb DNA fragment from the S. coelicolor chromosome cloned on a multicopy plasmid. Southern analysis, restriction map comparisons, and map locations of the cloned genes revealed that these genes were different from other known S. coelicolor genes concerned with actinorhodin biosynthesis or its pleiotropic regulation. Computer analysis of the DNA sequence showed five putative open reading frames (ORFs), which were named ORFA, ORFB, and ORFC (transcribed in one direction) and ORFD and ORFE (transcribed in the opposite direction). Subcloning experiments revealed that ORFB together with 137 bp downstream of it is responsible for antibiotic overproduction in S. lividans. Insertion of a phi C31 prophage into ORFB by homologous recombination gave rise to a mutant phenotype in which the production of actinorhodin, undecylprodigiosin, and the calcium-dependent antibiotic (but not methylenomycin) was reduced or abolished. The nonproducing mutants were not affected in the timing or vigor or sporulation. A possible involvement of ORFA in antibiotic production in S. coelicolor is not excluded. abaA constitutes a new locus which, like the afs and abs genes previously described, pleiotropically regulates antibiotic production. DNA sequences that hybridize with the cloned DNA are present in several different Streptomyces species.     

Streptomyces sp. Z-11-6, a novel producer of extracellular L-glutamate oxidase

Glutamate oxidase activity was studied in 1254Streptomyces strains isolated from the zonal soils of various regions of Russia and other countries. Seven strains proved to be producers of extracellular L-glutamate oxidase. The most active producer strain was identified, and the conditions of enzyme biosynthesis were optimized. A multistep mutagenesis-selection procedure allowed a genetically stable strain,Streptomyces sp. Z-11-6, to be obtained, whose glutamate oxidase activity was 40 times higher than that of the original natural isolate.     

响应面法优化海洋放线菌Streptomyces sp.YT-10抗菌物质的发酵条件

以啤酒酵母为指示菌,研究了海洋放线菌YT-10产抗菌物质的发酵条件。利用Plackett-Burman设计对影响YT-10产抗菌物质的因素的效应进行评价,筛选出具有显著效应的三个因素:葡萄糖、黄豆粉和氯化钠。利用响应面中心旋转组合设计优化三个显著因素,确定了葡萄糖,黄豆粉和氯化钠浓度分别为0.7%,1.3%,0.952%。优化后抑菌圈直径可达32.6mm,比原来增加了34.7%。     

Bluemomycin,a new naphthoquinone derivative from Streptomyces sp. with antimicrobial and cytotoxic properties

Biotechnology Letters - Streptomyces is one of the most prolific producers of economically important bioactive compounds used against several illnesses; it has also been found to produce...     

Mechanism of the incidental production of a melanin-like pigment during 6-demethylchlortetracycline production in Streptomyces aureofaciens

The secondary metabolite 6-demethylchlortetracycline (6-DCT), which is produced by Streptomyces aureofaciens, is used as a precursor of semisynthetic tetracyclines. Strains that produce 6-DCT also produce a melanin-like pigment (MP). The correlation between MP production and 6-DCT production was investigated by using S. aureofaciens NRRL 3203. Production of both MP and 6-DCT was repressed by phosphate or ammonium ions, suggesting that syntheses of these compounds are controlled by the same regulators. Ten chlortetracycline-producing recombinants were derived from 6-DCT-producing mutant NRRL 3203 by gene replacement. All of the recombinants produced chlortetracycline but not MP, indicating that MP production is the results of a defect in the 6-methylation step and suggesting that the polyketide nonaketideamide is a common intermediate leading to MP as well as 6-DCT. To further examine the possibility that MP might be synthesized via the 6-DCT-biosynthetic pathway, mutants defective in 6-DCT biosynthesis were derived from a 6-DCT producer. Some of these mutants were able to produce MP, while others, including mutants with mutations in the gene encoding anhydrotetracycline oxygenase, an enzyme catalyzing the penultimate step in the pathway, produced neither 6-DCT nor MP. Production of 6-DCT and production of MP were restored simultaneously by integrative transformation with the corresponding 6-DCT-biosynthetic genes, indicating that some of 6-DCT-biosynthetic enzymes are indispensable for MP production. These findings suggest that a defect in the 6-methylation step results in redirection of carbon flux from a certain intermediate in the 6-DCT-biosynthetic pathway to a shunt pathway and results in MP production.     

Acid-precipitable polymeric lignin from hardwood lignocellulose: production by a Streptomyces sp.

A Streptomyces sp. isolate, from decayed wood shavings, solubilized lignocellulose (LC) and lignin of Pinus radiata, producing about 50 mg acid-precipitable polymeric lignin per g LC. The product was poor in protein and carbohydrates and contained mainly vanillin, guaicol, vanillic and ferulic acids. Hardwood LC is thus suitable for producing APPL as a phenolic chemical feedstock.V.M. Kaluskar is with the Department of Microbiology, J and J Science College, Nadiad 387001, Gujarat, India. B.P. Kapadanis is with the Department of Microbiology, School of Sciences, University of Pune, Ganesh Khind, Pune-41107, Maharashtra, India. M.J. Penninckx is with the Unit of Microbial Physiology and Ecology, Free University of Brussels, c/o IPB 642, rue Engeland, B-1180, Brussels, Belgium     

Draft genome sequence of Streptomyces sp. strain Wigar10, isolated from a surface-sterilized garlic bulb

Streptomyces sp. strain Wigar10 was isolated from a surface-sterilized garlic bulb (Allium sativum var. Purple Stripe). Its genome encodes several novel secondary metabolite biosynthetic gene clusters and provides a genetic basis for further investigation of this strain's chemical biology and potential for interaction with its garlic host.     

Ansaetherone,a New Radical Scavenger from Streptomyces sp.

We identified a new radical scavenger, ansaetherone (C₂₆H₃₃NO₇), from a culture of the Streptomyces sp. USF-4727 strain. In our previous study, it was shown that this strain produced four lipoxygenase inhibitors, tetrapetalones A, B, C and D. The chemical structure of ansaetherone was elucidated by the spectroscopic method, indicating that this compound was constructed with an aglycon and a sugar moiety. This chemical structure suggested that ansaetherone was related to the tetrapetalones. This finding provided information regarding tetrapetalone biosynthesis. Ansaetherone showed radical scavenging activity with an ED₅₀ value of 300 μM in our assay.     

Cloning and characterization of a Streptomyces single module type non-ribosomal peptide synthetase catalyzing a blue pigment synthesis

In the present study, we cloned a gene, designated bpsA, which encodes a single module type non-ribosomal peptide synthetase (NRPS) from a D-cycloserine (DCS)-producing Streptomyces lavendulae ATCC11924. A putative oxidation domain is significantly integrated into the adenylation domain of the NRPS, and the condensation domain is absent from the module. When S. lividans was transformed with a plasmid carrying bpsA, the transformed cells produced a blue pigment, suggesting that bpsA is responsible for the blue pigment synthesis. However, to produce the blue pigment in Escherichia coli, the existence of the 4'-phosphopantetheinyl transferase (PPTase) gene from Streptomyces was necessary, in addition to bpsA. The chemical structure of the pigment was determined as 5,5'-diamino-4,4'-dihydroxy-3,3'-diazadiphenoquinone-(2,2'), called indigoidine. The bpsA gene product, designated BPSA, was overproduced in an E. coli host-vector system and purified to homogeneity, demonstrating that the recombinant enzyme prefers L-Gln as a substrate. The in vitro experiment using L-Gln also showed that the blue pigment was formed by the purified BPSA only when the enzyme was phosphopantetheinylated by adding a Streptomyces PPTase purified from E. coli cells. Each site-directed mutagenesis experiment of Lys(598), Tyr(601), Ser(603), and Tyr(608), which are seen in the oxidation domain of BPSA, suggests that these residues are essential for the binding of FMN to the protein and the synthesis of the blue pigment.     

Purification, Characterization, and Substrate Specificities of Multiple Xylanases from Streptomyces sp. Strain B-12-2

The endoxylanase complex from Streptomyces sp. strain B-12-2 was purified and characterized. The organism forms five distinct xylanases in the absence of significant cellulase activity when grown on oat spelt xylan. This is the largest number of endoxylanases yet reported for a streptomycete. On the basis of their physiochemical characteristics, they can be divided into two groups: the first group (xyl 1a and xyl 1b) consists of low-molecular-mass (26.4 and 23.8 kDa, respectively) neutral- to high-pI (6.5 and 8.3, respectively) endoxylanases. Group 1 endoxylanases are unable to hydrolyze aryl-beta-d-cellobioside, have low levels of activity against xylotetraose (X(4)) and limited activity against xylopentaose, produce little or no xylose, and form products having a higher degree of polymerization with complex substrates. These enzymes apparently carry out transglycosylation. The second group (xyl 2, xyl 3, and xyl 4) consists of high-molecular-mass (36.2, 36.2, and 40.5 kDa, respectively), low-pI (5.4, 5.0, and 4.8, respectively) xylanases. Group 2 endoxylanases are able to hydrolyze aryl-beta-d-cellobioside, show higher levels of activity against X(4), and hydrolyze xylopentaose completely with the formation of xylobiose and xylotriose plus limited amounts of X(4) and xylose. The enzymes display intergroup synergism when acting on kraft pulp. Despite intragroup similarities, each enzyme exhibited a unique action pattern and physiochemical characteristic. xyl 2 was highly glycosylated, and xyl 1b (but no other enzyme) was completely inhibited by p-hydroxymercuribenzoate.