Cloning of a chloramphenicol acetyltransferase gene of Streptomyces acrimycini and its expression in Streptomyces and Escherichia coli

Gil, José A.; Kieser, Helen M.; Hopwood, David A.

doi:10.1016/0378-1119(85)90197-0

Cited by 26 publications

(7 citation statements)

References 26 publications

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“…Instead, S. lividans 1326 and M252 modified the antibiotic by 0-acetylation. The inferred presence of CAT activity supports observations of weak CAT activity in strains M252 (Gil & Hopwood, 1985) and 1326 (W. Dittrich & H. Schrempf, unpublished), and would seem to conflict with earlier reports of its absence from strain 1326 (Shaw & Hopwood, 1976;Gil & Hopwood, 1985). However, 0-acetylation of chloramphenicol by this strain depends on the composition of the growth medium and the age of cultures (N. P. Ranade, unpublished) and it is possible that CAT activity is catabolite-repressed under some conditions.…”

Section: Discussionsupporting

confidence: 87%

Chloramphenicol resistance in Streptomyces: cloning and characterization of a chloramphenicol hydrolase gene from Streptomyces venezuelae

Mosher

Ranade

Schrempf

et al. 1990

Journal of General Microbiology

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A 6.5 kb DNA fragment containing a chloramphenicol-resistance gene of Streptomyces venezuelae ISP5230 was cloned in Streptomyces fitlidans M252 using the high-copy-number plasmid vector pIJ702. The gene was located within a 2.4 kb KpnI-SstI fragment of the cloned DNA and encoded an enzyme (chloramphenicol hydrolase) that catalysed removal of the dichloroacetyl moiety from the antibiotic. The deacylated product, p-nitrophenylserinol, was metabolized to p-nitrobenzyl alcohol and other compounds by enzymes present in S. f i v i h s M252. Examination of the genomic DNA from several sources using the cloned 6.5 kb SstI fragment from S. venezuefue ISP5230 as a probe showed a hybridizing region in the DNA from S. venezuelae 13s but none in the DNA from another chloramphenicol producer, Streptomycesphaeochromogenes NRRLB 3559. The resistance phenotype was not expressed when the 6.5 kb SstI fragment or a subfragment was subcloned behind the lac-promoter of plasmid pTZ18R in Escherichia cofi.

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Section: Discussionsupporting

confidence: 87%

Chloramphenicol resistance in Streptomyces: cloning and characterization of a chloramphenicol hydrolase gene from Streptomyces venezuelae

Mosher

Ranade

Schrempf

et al. 1990

Journal of General Microbiology

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“…No chemical investigation of the S. acrimycini secondary metabolites has been previously reported, but only genetic and morphological descriptions. [8][9][10][11] Although there is abundant literature data about dipeptides isolated from microbial sources, 12,16 their true origin remains controversial. Some authors suggest that dipeptides are fermentation artifacts, generated by hydrolysis of proteins present in the growth media.…”

Section: Resultsmentioning

confidence: 99%

Dipeptide metabolites from the marine derived bacterium Streptomyces acrymicini

Hernandez¹,

Macedo²,

Berlinck³

et al. 2004

J. Braz. Chem. Soc.

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A investigação química do extrato bruto obtido a partir do meio de crescimento do actinomiceto Streptomyces acrimycini isolado de sedimentos marinhos levou ao isolamento de dois dipeptídeos: a 8-amino-[1,4]diazonano-2,5-diona (1) e leucil-4-hidroxiprolina (3). Os compostos foram isolados a partir do seu meio de crescimento por uma série de etapas cromatográficas e identificados pela análise de seus dados espectroscópicos. O esqueleto macrocíclico da 8-amino-[1,4]diazonano-2,5-diona foi descrito apenas uma vez nas marinobactinas, sideróforos isolados a partir de uma bactéria marinha do gênero Marinobacter.The chemical investigation of the crude extract obtained from the growth media of the marinederived actinomycete Streptomyces acrimycini isolated from marine sediments led to the isolation of two new dipeptide derivatives: 8-amino-[1,4]diazonane-2,5-dione (1) and leucyl-4-hydroxyproline (3). The dipeptides were isolated from the growth media by a series of chromatographic steps, and identified by analysis of spectroscopic data. The macrocyclic carbon backbone of 8-amino-[1,4]diazonane-2,5-dione has been previously reported only once in marinobactins, siderophores isolated from the marine bacterium Marinobacter sp.

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“…We confirmed that three Streptomyces species had diacetyl chloramphenicol esterase activities, but these were at lower levels than those we observed with S. aurantia. This lower activity in the streptomycetes may explain why CAT has provided resistance in some Streptomyces (11). The streptomycetes, like S. aurantia, inhabit environments containing toxins as well as complex nutrients.…”

Section: Discussionmentioning

confidence: 99%

Spirochaeta aurantia Has Diacetyl Chloramphenicol Esterase Activity

Sohaskey

Barbour

2000

J Bacteriol

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The free-living spirochete Spirochaeta aurantia was nearly as susceptible to diacetyl chloramphenicol, the product of chloramphenicol acetyltransferase, as it was to chloramphenicol itself. This unexpected susceptibility to diacetyl chloramphenicol was wholly or partly the consequence of intrinsic carboxylesterase activity, as indicated by high-performance liquid chromatography, thin-layer chromatography, and microbiological assays. The esterase converted the diacetate to chloramphenicol, thus inhibiting spirochete growth. The esterase activity was cell associated, reduced by proteinase K, eliminated by boiling, and independent of the presence of either chloramphenicol or diacetyl chloramphenicol. S. aurantia extracts also hydrolyzed other esterase substrates, and two of these, ␣-napthyl acetate and 4-methylumbelliferyl acetate, identified an esterase of approximately 75 kDa in a nondenaturing gel. Carboxylesterases occur in Streptomyces species, but in this study their activity was weaker than that of S. aurantia. The S. aurantia esterase could reduce the effectiveness of cat as either a selectable marker or a reporter gene in this species.The antibiotic chloramphenicol blocks translation by interacting with the peptidyl transferase centers of ribosomes (25). Acquired resistance to chloramphenicol in eubacteria is most commonly provided by the enzyme chloramphenicol acetyltransferase (CAT), which is encoded by one of several different types of cat genes (reviewed in reference 27). Some CAT proteins also provide resistance to fusidic acid and crystal violet, but they do so by sequestering these compounds (3,26). CAT acetylates chloramphenicol once or twice; neither the monoacetate nor the diacetate form of chloramphenicol has been shown to have antibiotic activity (27). Most of the studies of chloramphenicol and CAT have been carried out with either gram-negative or gram-positive bacteria. Little is known about the activities of chloramphenicol and CAT in spirochetes, which are distinct from other bacteria in several characteristics (24,32).Spirochaeta aurantia is a pigmented, free-living spirochete found in aquatic environments. In comparison to most other known spirochetes, S. aurantia has simple growth requirements and a fast doubling time (5). These features make it a suitable model organism for genetic studies of spirochetes, and accordingly, we began development of a genetic system for S. aurantia. We had previously shown that a CAT gene of gram-positive bacteria could be expressed in transfected Borrelia burgdorferi (28, 30), and S. aurantia was known to be susceptible to chloramphenicol (4). Thus, there was reason to expect that the CAT gene would provide for positive selection and function as a reporter in S. aurantia as well. However, in preliminary experiments we found that S. aurantia was not only susceptible to chloramphenicol it was also unexpectedly susceptible to diacetyl chloramphenicol, the product of CAT.In the present study we characterized this phenomenon in more detail and investigated the ...

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Cloning of a chloramphenicol acetyltransferase gene of Streptomyces acrimycini and its expression in Streptomyces and Escherichia coli

Cited by 26 publications

References 26 publications

Chloramphenicol resistance in Streptomyces: cloning and characterization of a chloramphenicol hydrolase gene from Streptomyces venezuelae

Chloramphenicol resistance in Streptomyces: cloning and characterization of a chloramphenicol hydrolase gene from Streptomyces venezuelae

Dipeptide metabolites from the marine derived bacterium Streptomyces acrymicini

Spirochaeta aurantia Has Diacetyl Chloramphenicol Esterase Activity

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