Metabolic pathways in Microbispora sp. ATCC-PTA 5024, producer of NAI-107 lantibiotic

Proteomic study of the antibiotic producing Amycolatopsis balhimycina

From microbial proteomics to synthetic biology: Amycolatopsis balhimycina case

Actinomycetes, filamentous Gram-positive bacteria, are usually exploited as bio-farms naturally producing a wide range of small biologically active metabolites, such as antibiotics, extensively used in medicine, food-industry, chemistry and bio-remediation strategies. The development of high throughput technologies, like proteomics, allows functional genomic studies aimed at shedding light on molecular mechanisms controlling the production of useful compounds and macromolecules. Differential proteomic analyses, performed by using Two Dimensional PolyAcrylamide Gel Electrophoresis (2D-PAGE) coupled to mass spectrometry (MS) procedures, revealed novel links between balhimycin production (a va…

THE SMALL PROTEIN SCO2038 CONTROLS TRYPTOPHAN BIOSYNTHESIS AND DIFFERENTIATION IN STREPTOMYCES COELICOLOR

The cellular regulatory factors comprise regulatory proteins, small RNA and small proteins. It is known that the product of small orfs (smorfs) can regulate the translation of downstream elements and also can encode functional peptides involved in the regulation of specific pathways (Ladoukakis E. et al.,2011). In particular, in the model streptomycete Sfreptomyces coelicolor, smorfs (about 100-300 nucleotides) were identified in some amino acid biosynthetic gene clusters such as in the tryptophan trpCXBA locus (Limauro D. et al., I 990; Hu DS. et al., 1999). In S. coelicolor the molecular mechanisms that regulate tryptophan (Trp) biosynthesis are poorly understood and, unlike the trp opero…

Multi-omics of Pseudoalteromonas haloplanktis TAC125: a quest for antimicrobial metabolic pathways

Backgrounds The Antarctic strain Pseudoalteromonas haloplanktis TAC125 is one of the model organisms of cold-adapted bacteria and is currently exploited as a new alternative expression host for numerous biotechnological applications. Interestingly, this bacterium has been reported to be able to inhibit the growth of Burkholderia cepacia complex (Bcc) strains, opportunistic pathogens responsible for the infection of immune-compromised patients. Most likely, this occurs through the synthesis of several different compounds, including Volatile Organic Compounds (VOCs), whose nature and characteristics are currently mostly unknown. Objectives To obtain a complete picture of cellular processes di…

Genetic manipulation of spiramycin-producing strain Streptomyces ambofaciens ATCC 23877 by φC31 Att/Int system-based vectors results in a significant reduction of antibiotic production

The genus Streptomyces produces about two-thirds of naturally occurring antibiotics and many other biologically active secondary metabolites. The genetic manipulation of Streptomycetes is often labor and time intensive due to their large genome and complex development. Much progress has been made to develop gene transfer methods useful to construct antibiotic-producing strains with improved properties. The φC31 Att/Int system is an integration system that has been widely used to produce stable recombinants in Actinomycetes and its integration site is an attB site located in a pirin-like gene (pirA) of the bacterial chromosome. In this study we demonstrate that the integration of φC31 DNA in…

The small protein SCO2038 controls Streptomyces coelicolor differentiation by modulating tryptophan biosynthesis

Background In Streptomyces coelicolor amino acid metabolism is an important clue of the morphological and physiological differentiation program and, differently from other bacteria, the expression of amino acid biosynthetic genes is not subjected to endproduct negative regulation. In some amino acid biosynthetic gene clusters, such as tryptophan, histidine and proline, small orfs (about 100-300 nucleotides) were identified. These small orfs, such as sco2038, encode proteins whose cellular role have to be elucidated to highlight possible novel and crucial molecular mechanisms controlling amino acid synthesis and, thus, differentiation program. Objectives The aims of this work are: 1. the und…

trpX: a small orf involved in S. coelicolor tryptophan metabolism

The small protein TrpM modulates morpho-physiological differentiation in Streptomyces coelicolor

TrpM, a small protein of 63 amino acids, is encoded by a gene of the trpCMBA locus involved in tryptophan biosynthesis in the model actinomycete Streptomyces coelicolor. Indeed, the trpM knock-out mutant strain is characterized by a delayed growth on minimal medium, smaller aerial hyphae, and reduction of both spore and antibiotic actinorhodin production in comparison with the wild-type strain. These observations are in agreement with proteomic analyses which highlighted a role for TrpM in controlling i) tryptophan production through precursor availability and, thus ii) bacterial growth and morpho-physiological differentiation. To further elucidate the role of TrpM, a S. coelicolor trpM kno…

Omics approaches to elucidate the molecular physiology of lantibiotc NAI-107 production in Microbispora ATCC-PTA-5024

The small protein SCO2038 modulates tryptophan biosynthesis and morpho-physiological differentiation in Streptomyces coelicolor

In Streptomyces coelicolor small open reading frames were identified in several amino acids biosynthetic gene clusters, like SCO2038 (trpX) in the tryptophan trpCXBA locus. Here, the role of SCO2038, encoding a 63 amino acid protein, was investigated by both phenotypic and molecular analyses. A SCO2038 knockout mutant strain showed a delayed growth on minimal medium (MM), compromised actinorhodin biosynthesis and poor sporulation. The capability of this mutant to grow on MM was restored by tryptophan’s and its precursors’ supplementation. Pulldown and bacterial two hybrid assays revealed SCO2038 interaction with PepA, which is putatively involved in the metabolism of serine, glycine and cys…

Proteomics to elucidate the molecular physiology of Microbispora sp. ATCC-PTA-5024, the producer of NAI-107, a very promising lantibiotic

The filamentous actinobacterium Microbispora sp. ATCC-PTA-5024 produces the lantibiotic NAI-107 (Maffioli et al., 2014), which is effective against multidrug-resistant Gram-positive pathogens (Jabés et al., 2011). In actinomycetes, the biosynthesis of antibiotics is generally elicited as a physiological response controlled by a complex regulatory network involving global regulators, playing pleiotropic roles, and pathway-specific regulators, which activate the biosynthesis of biologically active molecules (Bibb, 2005)