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Journal of Basic Microbiology Apr 2021This study focuses on analyzing the protein expression pattern of intracellular proteins when Pseudomonas mendocina SMSKVR-3 exposed to 300 mM of arsenate to find out...
This study focuses on analyzing the protein expression pattern of intracellular proteins when Pseudomonas mendocina SMSKVR-3 exposed to 300 mM of arsenate to find out the proteins that are overexpressed or exclusively expressed in response to arsenate. The sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of protein expression at different time intervals showed the highest number of protein bands (14) that are overexpressed at 8 h of the time interval. It was also observed that treatment with at least 200 mM of As(V) is required to induce a difference in protein expression. Two-dimensional (2D)-PAGE analysis of 8-h sample exhibited 146 unique spots, 45 underexpressed, and 46 overexpressed spots in arsenate-treated sample. Based on the highest percent volume and fold change, three unique spots and one overexpressed spot were selected and analyzed by matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF/TOF) mass spectrometry (MS) analysis followed by the MASCOT search. These proteins were identified as ribosome-recycling factor (20.13 kDa), polyphosphate:ADP/GDP phosphotransferase (40.88 kDa), ribonuclease P protein component (14.96 kDa) and cobalt-precorrin-5B C(1)-methyltransferase (38.43 kDa) with MASCOT score of 54, 81, 94, and 100, respectively. All of these proteins help the bacteria to overcome arsenate stress.
Topics: Arsenates; Bacterial Proteins; Electrophoresis, Gel, Two-Dimensional; Electrophoresis, Polyacrylamide Gel; Pseudomonas mendocina; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
PubMed: 33448070
DOI: 10.1002/jobm.202000671 -
The Israel Medical Association Journal... Jun 2011
Topics: Adult; Anti-Bacterial Agents; Diagnosis, Differential; Hematologic Tests; Humans; Male; Microbial Sensitivity Tests; Pseudomonas Infections; Pseudomonas mendocina; Sepsis
PubMed: 21809738
DOI: No ID Found -
Nefrologia : Publicacion Oficial de La... 2017
Topics: Humans; Male; Peritoneal Dialysis; Peritonitis; Pseudomonas Infections; Pseudomonas mendocina; Young Adult
PubMed: 28655401
DOI: 10.1016/j.nefro.2016.11.004 -
Applied Microbiology and Biotechnology Feb 2019Polyhydroxyalkanoates (PHAs) can be produced by microorganisms from renewable resources and are regarded as promising bioplastics to replace petroleum-based plastics. A...
Polyhydroxyalkanoates (PHAs) can be produced by microorganisms from renewable resources and are regarded as promising bioplastics to replace petroleum-based plastics. A medium-chain-length PHAs (mcl-PHA)-producing strain Pseudomonas mendocina NK-01 was isolated previously by our lab and its whole-genome sequence is currently available. Morphology engineering of manipulating cell morphology-related genes has been applied for enhanced accumulation of the intracellular biopolymer short-chain-length PHAs (scl-PHA). However, it has not yet been reported to improve the yield of mcl-PHA by morphology engineering so far. In this work, several well-characterized cell morphology-related genes, including the cell fission ring (Z-ring) location genes minCD, peptidoglycan degradation gene nlpD, actin-like cytoskeleton protein gene mreB, Z-ring formation gene ftsZ, and FtsZ inhibitor gene sulA, were intensively investigated for their impacts on the cell morphology and mcl-PHA accumulation by gene knockout and overexpression in P. mendocina NKU, a upp knockout mutant of P. mendocina NK-01. For a minCD knockout mutant P. mendocina NKU-∆minCD, the average cell length was obviously increased and the mcl-PHA production was improved. However, the nlpD knockout mutant had a shorter cell length and lower mcl-PHA yield compared with P. mendocina NKU. Overexpression of mreB in P. mendocina NKU resulted in spherical cells. When ftsZ was overexpressed in P. mendocina NKU, the cell division was accelerated and the mcl-PHA titer was improved. Furthermore, mreB, ftsZ, or sulA was overexpressed in P. mendocina NKU-∆minCD. Consequently, the mcl-PHA titers were all increased compared with P. mendocina NKU-∆minCD carrying the empty vector. The multiple fission pattern was finally achieved in ftsZ-overexpressing NKU-∆minCD. In this work, improved production of mcl-PHA in P. mendocina NK-01 has been achieved by morphology engineering. This work provides an alternative strategy to enhance mcl-PHA accumulation in mcl-PHA-producing strains.
Topics: Gene Deletion; Gene Expression; Metabolic Engineering; Polyhydroxyalkanoates; Pseudomonas mendocina
PubMed: 30610286
DOI: 10.1007/s00253-018-9546-8 -
3 Biotech Sep 2019This study aimed to investigate the effects of cytoskeleton protein MreB on bacterial cell morphology and the synthesis of alginate oligosaccharides (AO) and...
This study aimed to investigate the effects of cytoskeleton protein MreB on bacterial cell morphology and the synthesis of alginate oligosaccharides (AO) and polyhydroxyalkanoate (PHA) by NK-01. To overexpress the gene, an expression vector encoding MreB-GFP fusion protein was constructed. The scanning electron microscope (SEM) showed that cells expressing MreB were longer than the wild ones, which agrees with MreB's relationship with the synthesis of peptidoglycan. Cells expressing the MreB-GFP fusion protein emitted green fluorescence under a fluorescence microscope, suggesting that MreB was functionally expressed in strain NK-01. Under a confocal laser scanning microscope, MreB was observed as located around the cell membrane. Furthermore, the recombinant strain could synthesize 0.961 g/L AO, which was 5.86-fold higher than wild-type strain. Through the medium optimization test, we finally selected the addition of 20 g/L glucose as the optimal glycogen addition for AO fermentation based on a high AO yield and high substrate transformation efficiency. The results indicated that overexpression of MreB affected the cell morphology, the activity of AO polymerase, and the efficiency of AO secretion. However, the synthesis of PHA for recombinant strain was slightly reduced. The results suggested that the overexpression of this cytoskeleton protein affected the yield of specific intracellular and extracellular products.
PubMed: 31497462
DOI: 10.1007/s13205-019-1873-7 -
Bioresource Technology Feb 2022Two biosafety strains, identified as Pseudomonas mendocina S16 and Enterobacter cloacae DS'5, were isolated from freshwater aquaculture ponds and showed significant...
Nitrogen removal characteristics and potential application of the heterotrophic nitrifying-aerobic denitrifying bacteria Pseudomonas mendocina S16 and Enterobacter cloacae DS'5 isolated from aquaculture wastewater ponds.
Two biosafety strains, identified as Pseudomonas mendocina S16 and Enterobacter cloacae DS'5, were isolated from freshwater aquaculture ponds and showed significant heterotrophic nitrification-aerobic denitrification abilities. Within 48 h, the inorganic nitrogen removal efficiencies in the two strains were 66.59 %-97.97 % (S16) and 72.27 %-96.44 % (DS'5). The optimal conditions for organic nitrogen removal of the two strains were temperature 20-35 °C and carbon/nitrogen (C/N) ratio 10-20 while using sodium citrate as the carbon source. Sequence amplification demonstrated the presence of the denitrification genes in both the two strains, and quantitative real-time PCR results showed that the coupled expression of nap + nar would improve the nitrate removal rate in S16. The nitrogen removal efficiencies of the two strains in immobilization culture systems were 79.80 %-98.58 % (S16) and 60.80 %-98.40 % (DS'5). This study indicated the great potential application of the two strains in aquaculture tail water treatment.
Topics: Aerobiosis; Aquaculture; Bacteria; Denitrification; Enterobacter cloacae; Heterotrophic Processes; Nitrification; Nitrites; Nitrogen; Ponds; Pseudomonas mendocina; Wastewater
PubMed: 34910970
DOI: 10.1016/j.biortech.2021.126541 -
Current Microbiology Nov 2020Even though organisms with squalene hopene cyclase activity involved in hopanoid synthesis has been reported earlier, their existence along with carotenoid synthesis is...
Even though organisms with squalene hopene cyclase activity involved in hopanoid synthesis has been reported earlier, their existence along with carotenoid synthesis is rarely reported. Here, we report the existence of hopanoid and C carotenoid biosynthetic pathway in Pseudomonas mendocina, the squalene hopene cyclase producing endophyte of the medicinal plant Murraya koenigii. The enzyme squalene hopene cyclase from Pseudomonas mendocina is involved in the synthesis of dehydrosqualene-mediated alternate pathway for carotenoid biosynthesis. The hopanoids are involved in membrane stability and integrity, and the carotene chromophores are involved in the photo protection of the cell. The orange-colored C carotenoid pigment 4-4' diaponeurosporenic acid in the extracellular extract of Pseudomonas mendocina with squalene cyclase activity was detected by the combination of UV/Vis spectrometry, FTIR, and Mass Spectrometry. 4-4' diaponeurosporenic acid could be traced as the end product of the carotenoid pathway and belonged to the xanthophyll group of carotenoids.
Topics: Biosynthetic Pathways; Carotenoids; Lyases; Pseudomonas mendocina
PubMed: 32894325
DOI: 10.1007/s00284-020-02180-3 -
Chemosphere Mar 2023The high hydrophobicity of n-hexane is the main reason why it is difficult to be removed biologically. In this study, the effects of bamboo-charcoal modified by...
The high hydrophobicity of n-hexane is the main reason why it is difficult to be removed biologically. In this study, the effects of bamboo-charcoal modified by bimetallic Fe/Pd (BBC) on n-hexane biodegradation by Pseudomonas mendocina NX-1 (PM) was investigated. The n-hexane removal efficiency was increased in the presence of BC. The highest n-hexane removal efficiency at 90.0% was achieved at 0.05 g L BCE and 3 g L NH under pH 7.7 and 35 °C. Additionally, protein content (45.9 μg mL) and negative cell surface zeta potential (-26.4 mV) were increased during biodegradation process, with PM-BBC being 43.1 μg mL and 19.1 mV. Bacterial growth was improved and maximum cell surface hydrophobicity was obtained after 20 h, which was 59.4% higher than the control with PM-BBC (37.7%) or PM (16.1%), showing biodegradation products of 1-butanol and acetic acid. The results indicate that BBC improved n-hexane biodegradation efficiency by promoting bacterial growth, reducing cell zeta potential, exposing hydrophobic proteins, and increasing cell surface hydrophobicity of bacterial strain NX-1. This investigation suggests that BBC-enhanced biodegradation can be promising to treat n-hexane-containing gas.
Topics: Pseudomonas mendocina; Charcoal; Biodegradation, Environmental; Hexanes
PubMed: 36657580
DOI: 10.1016/j.chemosphere.2023.137897 -
3 Biotech Oct 2017Poly(3-hydroxybutyrate--4-hydroxybutyrate) (P(3HB--4HB)) is a biodegradable plastic that is extensively utilized in many fields. In this work, P(3HB--4HB) powder was...
Poly(3-hydroxybutyrate--4-hydroxybutyrate) (P(3HB--4HB)) is a biodegradable plastic that is extensively utilized in many fields. In this work, P(3HB--4HB) powder was degraded by for the preparation of low-molecular-mass (LMW) P(3HB--4HB). After degradation, the remaining P(3HB--4HB) powder was analyzed via gel permeation chromatography (GPC), differential scanning calorimetry (DSC), X-ray powder diffraction (XRD), Fourier transform infrared (FTIR) spectroscopy, and proton nuclear magnetic resonance (H NMR) spectroscopy. The degradation of P(3HB--4HB) by occurred in two stages: the fast degradation stage (0-8 h) and the slow degradation stage (8-24 h). GPC analysis showed that the molecular weight of P(3HB--4HB) gradually decreased with degradation time. After 24 h of degradation, the weight-average molecular weight of P(3HB--4HB) was reduced to 4-5 kDa. DSC and XRD analyses both verified that the degree of crystallinity decreased with prolonged degradation time. The melting temperature of the degraded powder, however, remained unchanged. FTIR and H NMR analyses of the degraded powder showed that no new material was produced during degradation. Thus, the degradation of P(3HB--4HB) by could be used to produce LMW P(3HB--4HB) for use in various applications, such as the synthesis of amphiphilic block copolymers.
PubMed: 28828288
DOI: 10.1007/s13205-017-0824-4 -
Infection, Genetics and Evolution :... Aug 2011A diversity of molecular translocation mechanisms, including various secretion systems, has been elaborated in host-bacterial interactions. The newly described type VI...
A diversity of molecular translocation mechanisms, including various secretion systems, has been elaborated in host-bacterial interactions. The newly described type VI secretion system (T6SS) appears to be involved in bacterial pathogenesis by acting as a nano-syringe, contributing in translocation of several effector-proteins into the eukaryotic host cell cytoplasm. Recent evidences revealed the involvement of T6SS machinery in inter-bacterial interactions. Several Pseudomonas species are found to harbour multiple and well organised T6SS loci, however, their genomic structural similarities as well as phylogenetic divergence suggest an independent evolution. Until now elementary evidence was provided for the presence of T6SS in the genomes of Pseudomonas entomophila (Pen), an aggressive insect pathogen as well as the human opportunistic pathogen Pseudomonas mendocina (Pme). In this report we evidenced by in silico genome mining along with bioinformatic analysis the presence of genes encoding for putative T6SS core components and secreted proteins in the sequenced Pen L48 and Pme ymp, strains and designated their putative promoters, sigma factors binding sites and various regulatory proteins. Moreover, we investigated the phylogenetic relatedness of four T6SS core proteins from these strains with their orthologues from various Pseudomonas species. Our analysis revealed two phylogenetically distinguishable T6SS loci in the genome of Pme that appeared to be highly homologous to Pseudomonas aeruginosa Hcp-Secretion Island-I (HSI-I) and -II. Our findings suggest that Pme could be excellent additional to P. aeruginosa model, for the elucidation of HSI-I and -II biological role(s), avoiding the overlapping activity HSI-III (Lesic et al., 2009), which is missing from Pme's genome. Likewise, our analysis revealed the presence of a unique entire T6SS in Pen genome, which appears to be phylogenetically close to Pme T6SS-II and P. aeruginosa HSI-II. Since Pen lacks the common secretion systems T3SS and T4SS, the single T6SS locus could have an enforced role in the insect-bacterial interactions, providing thus a promising model for studying its biological function.
Topics: Bacterial Proteins; Bacterial Secretion Systems; Base Composition; Computational Biology; Data Mining; Gene Expression Regulation, Bacterial; Gene Regulatory Networks; Genes, Bacterial; Models, Biological; Multigene Family; Phylogeny; Pseudomonas
PubMed: 21600307
DOI: 10.1016/j.meegid.2011.04.029