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273
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Introduction
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Publications
Publications (37)
The DNA repair endonuclease EndoMS/NucS is highly conserved in Archaea and Actinobacteria. This enzyme is able to recognize and cleave dsDNA carrying a mismatched base pair, and its activity is enhanced by the interaction with the sliding clamp of the replisome. Today, EndoMS/NucS has been established as the key protein of a non-canonical mismatch...
Some bacterial peptidyl-prolyl cis/trans isomerases (PPIases) are involved in secretory protein folding after the translocation step. Streptomyces lividans has been used as a host for engineering extracellular overproduction of homologous and heterologous proteins in industrial applications. Although the mechanisms governing the major secretory pat...
Background:
Streptomyces lividans is an appealing host for the production of proteins of biotechnological interest due to its relaxed exogenous DNA restriction system and its ability to secrete proteins directly to the medium through the major Sec or the minor Tat routes. Often, protein secretion displays non-uniform time-dependent patterns. Under...
Background:
Bacterial secretory proteins often require the formation of disulphide bonds outside the cell to acquire an active conformation. Thiol-disulphide oxidoreductases are enzymes that catalyse the formation of disulphide bonds. The bacterium Streptomyces lividans is a well-known host for the efficient secretion of overproduced homologous an...
Background:
Streptomyces lividans has demonstrated its value as an efficient host for protein production due to its ability to secrete functional proteins directly to the media. Secretory proteins that use the major Sec route need to be properly folded outside the cell, whereas secretory proteins using the Tat route appear outside the cell correct...
Gram-positive soil bacteria included in the genus Streptomyces produce a large variety of secondary metabolites in addition to extracellular hydrolytic enzymes. From the industrial and commercial viewpoints, the S. lividans strain has generated greater interest as a host bacterium for the overproduction of homologous and heterologous hydrolytic enz...
Proteins identified by nano LC–MS/MS Triple Tof analysis in the purified His6-CssR.
The Table indicates the number of Mascot protein score and the number of peptides identified for each protein by nano mass spectrometry analysis in the eluted fraction (E2. S1 Fig).
(DOCX)
Analysis of the expression of His6-CssR in E. coli strain BL21 (DE3).
A) E.coli cells overexpressing His6-CssR were grown and processed as described in Material and methods. The supernatant (S) containing the cytosol fraction was loaded onto a chromatography column filled with a Cobalt-containing resin. The concentration of the (S) fraction loaded...
Overproduction of Sec-proteins in S. lividans accumulates misfolded proteins outside of the cytoplasmic membrane where the accumulated proteins interfere with the correct functioning of the secretion machinery and with the correct cell functionality, triggering the expression in S. lividans of a CssRS two-component system which regulates the degrad...
Introduction:
Biological communities present in soil are essential to sustainable and productive agricultural practices; however, an accurate determination of the ecological status of agricultural soils remains to date an elusive task. An ideal indicator should be pervasive, play a relevant role in the ecosystem, show a rapid and proportional answ...
Preliminary statistical analysis.
The results of CCA and Indicator Value analyses obtained with each of the three classification methods are provided in separate subfolders as a compressed Zip archive.
(ZIP)
Taxonomic groups observed.
Details of all the taxonomic groups observed in the samples. The values reported are organized by taxonomic level and sample. Both, the frequencies (total number) of reads assigned to each taxonomic group and the percent that a group represents at each level in each sample are provided.
(XLS)
Statistical analysis of taxonomic changes observed.
The table contains the statistical analysis (G-test followed by post-hoc Fisher's exact and χ2 tests) of taxonomical changes observed between each sample subject to an external stress and its respective control. Changes have been classified by taxonomic level, experiment, stress applied and time o...
Extracellular protein production by Gram-positive bacteria, such as Streptomyces, may be complementary to current established protein production processes. The performance of a Streptomyces lividans mutant strain, deficient in the major signal peptidase (SipY) is investigated for the production of proteins secreted via the secondary Tat pathway. Th...
Streptomyces lividans uses mainly two pathways to target secretory proteins to the cytoplasmic membrane. The major pathway (Sec pathway) transports pre-proteins using the signal recognition particle, and the minor Tat pathway is responsible for the secretion using a folded conformation of a relatively low number of proteins. The signal peptides of...
Streptomyces lividans is considered an efficient host for the secretory production of homologous and heterologous proteins. To identify possible bottlenecks in the protein production process, a comparative transcriptomic approach was adopted to study cellular responses during the overproduction of a Sec-dependent model protein (alpha-amylase) and a...
Bacterial lipoproteins are a specialised class of membrane proteins that represent a small percentage of the proteome of Gram-positive bacteria, yet these lipoproteins have been reported to play important roles in nutrient scavenging, cell envelope assembly, protein folding, environmental signalling, host cell adhesion and virulence. Upon transloca...
Misfolded proteins accumulating outside the bacterial cytoplasmic membrane can interfere with the secretory machinery, hence the existence of quality factors to eliminate these misfolded proteins is of capital importance in bacteria that are efficient producers of secretory proteins. These bacteria normally use a specific two-component system to re...
Quantitative RT-PCR analysis of the two-component system and the three genes encoding HtrA-like proteases in the S. coelicolor DegU overproducer strain. Quantitative RT-PCR analysis of the two-component system and the HtrA- like protease genes in the S. coelicolor DegU overproducer strain (S.coelicolor M28; [26]) compared to that of the correspondi...
Deficiency in the translocase complex (SecG mutant strain) or in the major type I signal peptidase (SipY mutant strain) function in Streptomyces lividans resulted, as expected, in a drastic reduction of secretory protein production and in a bald phenotype. The transcriptional profiling of both strains showed that the expression of a set of genes in...
Background
Bacterial two-component signal transduction regulatory systems are the major set of signalling proteins frequently mediating responses to changes in the environment. They typically consist of a sensor, a membrane-associated histidine kinase and a cytoplasmic response regulator. The membrane-associated sensor detects the environmental sig...
Genes modulated by disruption of the SCO5784–5785 operon. Transcriptional units potentially ppGpp regulated [10] are underlined. *Genes down regulated in the strain carrying the SCO5785 gene in multicopy.
(DOC)
ICPL analysis of extracellular proteins of S. coelicolor M28 labelled with C12. Extracellular proteins of S. coelicolor M145 were labelled with C13 and those of S. coelicolor M28 were labelled with C12 at 24 h of growth.
(DOC)
Oligonucleotide primers used for gene transcript amplification. Oligonucleotide sequences start and terminate at their 5′ and 3′ ends, respectively.
(DOC)
Subtilisin inhibitor production and overall pattern of extracellular proteins. (A) Subtilisin inhibitor activity in cultures of S. coelicolor M28 (dark blocks) and its isogenic wild type strain (light blocks) grown in minimal medium. Values are given as a percentage of residual subtilisin activity in the assay. Total extracellular protein from expo...
ICPL analysis of extracellular proteins of S. coelicolor M28 labelled with C13. Extracellular proteins of S. coelicolor M145 were labelled with C12 and those of S. coelicolor M28 were labelled with C13 at 24 h of growth.
(DOC)
Expression of the steffimycin gene cluster in Steptomyces albus in combination with plasmids directing the biosynthesis of different neutral and branched-chain deoxyhexoses led to the identification of twelve new glycosylated derivatives of steffimycin with different degrees of decoration in the tetracyclic core. These experiments demonstrate the f...
The biosynthetic gene cluster for the aromatic polyketide steffimycin of the anthracycline family has been cloned and characterized
from “Streptomyces steffisburgensis” NRRL 3193. Sequence analysis of a 42.8-kbp DNA region revealed the presence of 36 open reading frames (ORFs) (one of them
incomplete), 24 of which, spanning 26.5 kb, are probably in...
