Additional file 4: of Elucidating the molecular physiology of lantibiotic NAI-107 production in Microbispora ATCC-PTA-5024

Supplementary Results section. (PDF 123 kb)

LAPTOP: Lantibiotic production, Technology, optimization and improved process

Omic-based strategies reveal novel links between primary metabolism and antibiotic production

biotecnologiche di batteri del suolo

AROMATIC AMINOACIDS AND ANTIBIOTIC PRODUCTION IN STREPTOMYCES COELICOLOR

Elucidating the molecular physiology of lantibiotic NAI-107 production in Microbispora ATCC-PTA-5024.

Background The filamentous actinomycete Microbispora ATCC-PTA-5024 produces the lantibiotic NAI-107, which is an antibiotic peptide effective against multidrug-resistant Gram-positive bacteria. In actinomycetes, antibiotic production is often associated with a physiological differentiation program controlled by a complex regulatory and metabolic network that may be elucidated by the integration of genomic, proteomic and bioinformatic tools. Accordingly, an extensive evaluation of the proteomic changes associated with NAI-107 production was performed on Microbispora ATCC-PTA-5024 by combining two-dimensional difference in gel electrophoresis, mass spectrometry and gene ontology approaches. R…

Inorganic phosphate is a trigger factor for Microbispora sp. ATCC-PTA-5024 growth and NAI-107 production

A putative membrane protein and an ABC-transporter of Planobispora rosea stimulate antibacterial activity in Streptomyces lividans

Expression in Streptomyces lividans of Nonomuraea genes cloned in an artificial chromosome

A bacterial artificial chromosomal library of Nonomuraea sp. ATCC39727 was constructed using Escherichia coli-Streptomyces artificial chromosome (ESAC) and screened for the presence of dbv genes known to be involved in the biosynthesis of the glycopeptide A40926. dbv genes were cloned as two large, partially overlapping, fragments and transferred into the host Streptomyces lividans, thus generating strains S. lividansColon, two colonsNmESAC50 and S. lividansColon, two colonsNmESAC57. The heterologous expression of Nonomuraea genes in S. lividans was successfully demonstrated by using combined RT-PCR and proteomic approaches. MALDI-TOF analysis revealed that a Nonomuraea ABC transporter is e…

Identification and characterization of new prolylendopeptidases (PEPs) from Actinomycetes

Applicazioni biotecnologiche di batteri del suolo

Regulation of antibiotic biosynthesis in Actinomycetes.

Amino acid biosynthesis in Streptomyces coelicolor

Dbv3 e Dbv4: due regolatori della biosintesi dell’antibiotico glicopeptidico A40926 di Nonomuraea ATCC39727

Additional file 5: of Elucidating the molecular physiology of lantibiotic NAI-107 production in Microbispora ATCC-PTA-5024

Figure S1-S6 with corresponding figure legends. (PDF 511 kb)

Additional file 5: of Elucidating the molecular physiology of lantibiotic NAI-107 production in Microbispora ATCC-PTA-5024

Figure S1-S6 with corresponding figure legends. (PDF 511 kb)

Aromatic aminoacids and Calcium Dependent Antibiotic:Relation of primary metabolism to antibiotic production in Actinomycetes

From tryptophan metabolism to peptide antibiotic production in Streptomyces coelicolor

The tryptophan is a precursor of the lipopeptide calcium dependent antibiotic (CDA), produced from Streptomyces coelicolor and closely related to important antibiotics such as daptomycin. We have focused our attention on the correlation between CDA production and tryptophan metabolism in order to identify new strategies aimed at increasing the production of peptide antibiotics.

Aminoacid metabolism and its impact on antibiotic production

TrpM, a Small Protein Modulating Tryptophan Biosynthesis and Morpho-Physiological Differentiation in Streptomyces coelicolor A3(2).

In the model actinomycete Streptomyces coelicolor A3(2), small open reading frames encoding proteins with unknown functions were identified in several amino acid biosynthetic gene operons, such as SCO2038 (trpX) in the tryptophan trpCXBA locus. In this study, the role of the corresponding protein in tryptophan biosynthesis was investigated by combining phenotypic and molecular analyses. The 2038KO mutant strain was characterized by delayed growth, smaller aerial hyphae and reduced production of spores and actinorhodin antibiotic, with respect to the WT strain. The capability of this mutant to grow on minimal medium was rescued by tryptophan and tryptophan precursor (serine and/or indole) su…

A putative membrane protein and an ABC-transporter of Planobispora rosea induce antibacterial activity of Streptomyces lividans

Regulation of the biosynthesis of the glycopeptide antibiotic A40926 in Nonomuraea ATCC39727

The actinomycete Nonomuraea produces the glycopeptide A40926, precursor of dalbavancin. Dalbavancin is a novel lipoglycopeptide agent against emerging resistant Gram-positive cocci with superior pharmacodynamics properties compared to vancomycin. Chemically, a glycopeptide antibiotic consists of a heptapeptide core constituted by proteinogenic and nonproteinogenic amino acids such as 3,5-dihydroxyphenylglycine (DPG) and 4-hydroxyphenylglycine (HPG). The heptapeptide is assembled by a nonribosomal peptide synthetase and modified by oxidative cross-linking of the electron-rich aromatic side chains, halogenation, sulfation, methylation, acylation, and glycosylation. The A40926 biosynthetic gen…

Two heterologously expressed Planobispora rosea proteins cooperatively induce Streptomyces lividans thiostrepton uptake and storage from the extracellular medium.

Abstract Background A bacterial artificial chromosomal library of Planobispora rosea, a genetically intractable actinomycete strain, was constructed using Escherichia coli-Streptomyces artificial chromosome (ESAC) and screened for the presence of genes known to be involved in the biosynthesis of antibiotics. Results One clone with a 40 kb insert showed antimicrobial activity against Gram positive bacteria. Insert sequence analysis and subcloning experiments revealed that the bioactivity was due to a 3.5 kb DNA fragment containing two open reading frames. These orfs encode two proteins with high similarity to a putative membrane protein of Streptomyces coelicolor and to the nogalamycin resis…

Proteomic investigations to unravel molecular physiology of the lantibiotic producer Microbispora sp. ATCC‐PTA‐5024

Tryptophan catabolism via kynurenine production in Streptomyces coelicolor: identification of three genes coding for the enzymes of tryptophan to anthranilate pathway

Most enzymes involved in tryptophan catabolism via kynurenine formation are highly conserved in Prokaryotes and Eukaryotes. In humans, alterations of this pathway have been related to different pathologies mainly involving the central nervous system. In Bacteria, tryptophan and some of its derivates are important antibiotic precursors. Tryptophan degradation via kynurenine formation involves two different pathways: the eukaryotic kynurenine pathway, also recently found in some bacteria, and the tryptophan-to-anthranilate pathway, which is widespread in microorganisms. The latter produces anthranilate using three enzymes also involved in the kynurenine pathway: tryptophan 2,3-dioxygenase (TD…

Additional file 4: of Elucidating the molecular physiology of lantibiotic NAI-107 production in Microbispora ATCC-PTA-5024

Supplementary Results section. (PDF 123 kb)

Impact of aminoacid metabolism on antibiotic production

A proteomic investigation to explore biochemical capabilities of a lantibiotic producer Microbispora strain

Impact of aminoacid metabolism on antibiotic production in Streptomyces

Stategie metaboliche indotte dal triptofano in Streptomyces coelicolor

AN ABC-TRANSPORTER AND A MEMBRANE PROTEIN OF PLANOBISPORA ROSEA INDUCE ANTIBACTERIAL ACTIVITY IN STREPTOMYCES LIVIDANS

Inorganic phosphate is a trigger factor for Microbispora sp. ATCC-PTA-5024 growth and NAI-107 production

Background NAI-107, produced by the actinomycete Microbispora sp. ATCC-PTA-5024, is a promising lantibiotic active against Gram-positive bacteria and currently in late preclinical-phase. Lantibiotics (lanthionine-containing antibiotics) are ribosomally synthesized and post-translationally modified peptides (RiPPs), encoded by structural genes as precursor peptides. The biosynthesis of biologically active compounds is developmentally controlled and it depends upon a variety of environmental stimuli and conditions. Inorganic phosphate (Pi) usually negatively regulates biologically-active molecule production in Actinomycetes, while it has been reported to have a positive control on lantibiotic…

Microbispora sp. proteomic analysis to study metabolic pathway changes during lantibiotic production

Regulation of the biosynthesis of dalbavancin

Having a look at Microbispora sp. ATCC-PTA-5024, a lantibiotic producer, "from the cradle to the grave" at the proteome level

Microbispora sp. ATCC-PTA-5024 proteomic analysis to study metabolic pathway changes before and during lantibiotic production

Heterologous expression of Nonomuraea sp. ATCC39727 genes in Streptomyces lividans

Identification of SCP2165, a new SCP2-derived plasmid of Streptomyces coelicolor A3(2).

Aims:  Characterization of SCP2165, a plasmid identified in the Gram-positive bacterium Streptomyces coelicolor A3(2). Methods and Results:  Pulsed-field gel electrophoresis (PFGE) of mycelia of a S. coelicolor strain embedded in low melting agarose revealed the presence of a plasmid. Restriction enzyme mapping and sequence analysis of a 2·1 kb fragment revealed that this plasmid could be SCP2. SCP2 and its spontaneous derivative SCP2* are self-transmissible plasmids and have chromosome mobilizing ability (c.m.a.). SCP2* has a c. 1000-fold increased c.m.a. compared with SCP2. Interestingly the plasmid, named SCP2165, shows a c.m.a. from 5 × 10−2 to 1 × 10−1 which is 50–100-fold higher than …

Additional file 2: Table S2. of Elucidating the molecular physiology of lantibiotic NAI-107 production in Microbispora ATCC-PTA-5024

Description, functional classification, abundance profile and mass spectrometry identification parameters of differentially represented Microbispora ATCC-PTA-5024 proteins identified from global proteome analysis at D substages. (XLS 107 kb)

Additional file 6: Table S5. of Elucidating the molecular physiology of lantibiotic NAI-107 production in Microbispora ATCC-PTA-5024

Description, abundance profile and mass spectrometry identification parameters of differentially represented spots containing multiple protein components. (XLS 45 kb)

Additional file 3: Table S3. of Elucidating the molecular physiology of lantibiotic NAI-107 production in Microbispora ATCC-PTA-5024

Description, functional classification, abundance profile and mass spectrometry identification parameters of differentially represented Microbispora ATCC-PTA-5024 proteins identified from membrane proteome analysis at A substages. (XLSX 37 kb)

Additional file 2: Table S2. of Elucidating the molecular physiology of lantibiotic NAI-107 production in Microbispora ATCC-PTA-5024

Description, functional classification, abundance profile and mass spectrometry identification parameters of differentially represented Microbispora ATCC-PTA-5024 proteins identified from global proteome analysis at D substages. (XLS 107 kb)

Additional file 6: Table S5. of Elucidating the molecular physiology of lantibiotic NAI-107 production in Microbispora ATCC-PTA-5024

Description, abundance profile and mass spectrometry identification parameters of differentially represented spots containing multiple protein components. (XLS 45 kb)

Additional file 7: Table S4. of Elucidating the molecular physiology of lantibiotic NAI-107 production in Microbispora ATCC-PTA-5024

Description, functional classification, abundance profile and mass spectrometry identification parameters of differentially represented proteins due to NAI-107 exposure in Microbispora ATCC-PTA-5024 RP0 strain. (XLSX 32 kb)

Additional file 1: Table S1. of Elucidating the molecular physiology of lantibiotic NAI-107 production in Microbispora ATCC-PTA-5024

Description, functional classification, abundance profile and mass spectrometry identification parameters of differentially represented Microbispora ATCC-PTA-5024 proteins identified from global proteome analysis at A substages. (XLSX 48 kb)

Additional file 8: Table S6. of Elucidating the molecular physiology of lantibiotic NAI-107 production in Microbispora ATCC-PTA-5024

Numbers of KEGG orthology groups participating in molecular and metabolic processes as inferred from genome and proteome analyses, respectively. (XLS 24 kb)

Additional file 3: Table S3. of Elucidating the molecular physiology of lantibiotic NAI-107 production in Microbispora ATCC-PTA-5024

Description, functional classification, abundance profile and mass spectrometry identification parameters of differentially represented Microbispora ATCC-PTA-5024 proteins identified from membrane proteome analysis at A substages. (XLSX 37 kb)

Additional file 7: Table S4. of Elucidating the molecular physiology of lantibiotic NAI-107 production in Microbispora ATCC-PTA-5024

Description, functional classification, abundance profile and mass spectrometry identification parameters of differentially represented proteins due to NAI-107 exposure in Microbispora ATCC-PTA-5024 RP0 strain. (XLSX 32 kb)

Additional file 8: Table S6. of Elucidating the molecular physiology of lantibiotic NAI-107 production in Microbispora ATCC-PTA-5024

Numbers of KEGG orthology groups participating in molecular and metabolic processes as inferred from genome and proteome analyses, respectively. (XLS 24 kb)

Additional file 1: Table S1. of Elucidating the molecular physiology of lantibiotic NAI-107 production in Microbispora ATCC-PTA-5024

Description, functional classification, abundance profile and mass spectrometry identification parameters of differentially represented Microbispora ATCC-PTA-5024 proteins identified from global proteome analysis at A substages. (XLSX 48 kb)