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Molecules (Basel, Switzerland) Dec 2020AJ003 produces glycosyl-4,4'-diaponeurosporen-4'-ol-4-oic acid as its main carotenoid. Five carotenoid pathway genes were presumed to be present in the genome of...
AJ003 produces glycosyl-4,4'-diaponeurosporen-4'-ol-4-oic acid as its main carotenoid. Five carotenoid pathway genes were presumed to be present in the genome of AJ003; however, 4,4-diaponeurosporene oxidase (CrtP) was non-functional, and a gene encoding aldehyde dehydrogenase (AldH) was not identified. In the present study, a genome mining approach identified two missing enzymes, CrtP2 and AldH2454, in the glycosyl-4,4'-diaponeurosporen-4'-ol-4-oic acid biosynthetic pathway. Moreover, CrtP2 and AldH enzymes were functional in heterologous and generated two carotenoid aldehydes (4,4'-diapolycopene-dial and 4,4'-diaponeurosporene-4-al) and two carotenoid carboxylic acids (4,4'-diaponeurosporenoic acid and 4,4'-diapolycopenoic acid). Furthermore, the genes encoding CrtP2 and AldH2454 were located at a distance the carotenoid gene cluster of .
Topics: Aldehyde Dehydrogenase; Carotenoids; Databases, Protein; Genome, Bacterial; Metabolic Networks and Pathways; Oxidoreductases; Planococcaceae
PubMed: 33322786
DOI: 10.3390/molecules25245892 -
Microbial Ecology May 2024Biocrust inoculation and microbially induced carbonate precipitation (MICP) are tools used in restoring degraded arid lands. It remains unclear whether the ecological...
Biocrust inoculation and microbially induced carbonate precipitation (MICP) are tools used in restoring degraded arid lands. It remains unclear whether the ecological functions of the two tools persist when these methods are combined and subjected to freeze-thaw (FT) cycles. We hypothesized a synergetic interaction between MICP treatment and biocrust under FT cycles, which would allow both components to retain their ecological functions. We grew cyanobacterial (Nostoc commune) biocrusts on bare soil and on MICP (Sporosarcina pasteurii)-treated soil, subjecting them to repeated FT cycles simulating the Mongolian climate. Generalized linear modeling revealed that FT cycling did not affect physical structure or related functions but could increase the productivity and reduce the nutrient condition of the crust. The results confirm the high tolerance of MICP-treated soil and biocrust to FT cycling. MICP treatment + biocrust maintained higher total carbohydrate content under FT stress. Our study indicates that biocrust on biomineralized soil has a robust enough structure to endure FT cycling during spring and autumn and to promote restoration of degraded lands.
Topics: Soil Microbiology; Soil; Freezing; Cyanobacteria; Carbonates; Ecosystem; Sporosarcina
PubMed: 38730059
DOI: 10.1007/s00248-024-02389-w -
Frontiers in Microbiology 2016Current diagnostic tests for Johne's disease (JD), a chronic granulomatous inflammation of the gastrointestinal tract of ruminants caused by Mycobacterium avium...
Current diagnostic tests for Johne's disease (JD), a chronic granulomatous inflammation of the gastrointestinal tract of ruminants caused by Mycobacterium avium subspecies paratuberculosis (MAP), lack the sensitivity to identify infected animals at early (asymptomatic) stages of the disease. The objective was to determine the pattern of MAP-associated dysbiosis of intestinal microbiota as a potential biomarker for early detection of infected cattle. To that end, genomic DNA was extracted from ileal mucosa and fecal samples collected from 28 MAP-positive and five control calves. High-throughput Illumina sequencing of the V4 hypervariable region of the 16S rRNA gene was used for community profiling of ileal mucosa-associated (MAM) or fecal microbiota. The PERMANOVA analysis of unweighted UniFrac distances revealed distinct clustering of ileal MAM (P = 0.049) and fecal microbiota (P = 0.068) in MAP-infected vs. control cattle. Microbiota profile of MAP-infected animals was further investigated by linear discriminant analysis effective size (LEfSe); several bacterial taxa within the phylum Proteobacteria were overrepresented in ileal MAM of control calves. Moreover, based on reconstructed metagenomes (PICRUSt) of ileal MAM, functional pathways associated with MAP infection were inferred. Enrichment of lysine and histidine metabolism pathways, and underrepresentation of glutathione metabolism and leucine and isoleucine degradation pathways in MAP-infected calves suggested potential contributions of ileal MAM in development of intestinal inflammation. Finally, simultaneous overrepresentation of families Planococcaceae and Paraprevotellaceae, as well as underrepresentation of genera Faecalibacterium and Akkermansia in the fecal microbiota of infected cattle, served as potential biomarker for identifying infected cattle during subclinical stages of JD. Collectively, based on compositional and functional shifts in intestinal microbiota of infected cattle, we inferred that this dynamic network of microorganisms had an active role in intestinal homeostasis.
PubMed: 27065983
DOI: 10.3389/fmicb.2016.00426 -
Microbial Cell Factories Nov 2018N-acylhomoserine lactones (AHLs) are well-studied signalling molecules produced by some Gram-negative Proteobacteria for bacterial cell-to-cell communication or quorum...
BACKGROUND
N-acylhomoserine lactones (AHLs) are well-studied signalling molecules produced by some Gram-negative Proteobacteria for bacterial cell-to-cell communication or quorum sensing. We have previously demonstrated the degradation of AHLs by an Antarctic bacterium, Planococcus versutus L10.15, at low temperature through the production of an AHL lactonase. In this study, we cloned the AHL lactonase gene and characterized the purified novel enzyme.
RESULTS
Rapid resolution liquid chromatography analysis indicated that purified AidP possesses high AHL-degrading activity on unsubstituted, and 3-oxo substituted homoserine lactones. Liquid chromatography-mass spectrometry analysis confirmed that AidP functions as an AHL lactonase that hydrolyzes the ester bond of the homoserine lactone ring of AHLs. Multiple sequence alignment analysis and phylogenetic analysis suggested that the aidP gene encodes a novel AHL lactonase enzyme. The amino acid composition analysis of aidP and the homologous genes suggested that it might be a cold-adapted enzyme, however, the optimum temperature is 28 °C, even though the thermal stability is low (reduced drastically above 32 °C). Branch-site analysis of several aidP genes of Planococcus sp. branch on the phylogenetic trees also showed evidence of episodic positive selection of the gene in cold environments. Furthermore, we demonstrated the effects of covalent and ionic bonding, showing that Zn is important for activity of AidP in vivo. The pectinolytic inhibition assay confirmed that this enzyme attenuated the pathogenicity of the plant pathogen Pectobacterium carotovorum in Chinese cabbage.
CONCLUSION
We demonstrated that AidP is effective in attenuating the pathogenicity of P. carotovorum, a plant pathogen that causes soft-rot disease. This anti-quorum sensing agent is an enzyme with low thermal stability that degrades the bacterial signalling molecules (AHLs) that are produced by many pathogens. Since the enzyme is most active below human body temperature (below 28 °C), and lose its activity drastically above 32 °C, the results of a pectinolytic inhibition assay using Chinese cabbage indicated the potential of this anti-quorum sensing agent to be safely applied in the field trials.
Topics: 4-Butyrolactone; Amino Acid Sequence; Antarctic Regions; Bacterial Proteins; Carboxylic Ester Hydrolases; Models, Molecular; Pectins; Pectobacterium carotovorum; Planococcus Bacteria; Quorum Sensing; Sequence Analysis, Protein; Substrate Specificity
PubMed: 30445965
DOI: 10.1186/s12934-018-1024-6 -
The Journal of General and Applied... May 2024Proteolytic enzymes stand out as the most widely employed category utilized in manufacturing industry. A new protease was separated from Planococcus sp.11815 strain and...
Proteolytic enzymes stand out as the most widely employed category utilized in manufacturing industry. A new protease was separated from Planococcus sp.11815 strain and named as nprS-15615 in this research. The gene of this protease has not been reported, and its enzymatic properties have been studied for the first time. To enhance enzyme production, the Planococcus sp. protease gene was expressed in Bacillus licheniformis 2709. The expression level of nprS-15615 was observed under the control of regulatory elements P. nprS-15615 protease activity reached 1186.24±32.87 U/mL after 48 hours of cultivation in shake flasks which was nearly four times the output of the original bacteria (291.38±25.73U/mL). The optimum temperature and pH of the recombinant protease were 30 ℃ and 8.0, respectively.The enzyme exhibited the highest capacity for hydrolyzing casein and demonstrated resilience towards a NaCl concentration of 10.0% (wt/v). Furthermore, in the presence of 0.5% surfactants, the recombinant protease activity can maintain above 75%, and with the existence of 0.5% liquid detergents, there was basically no loss of enzyme activity which indicated that nprS-15615 had good compatibility with surfactants and liquid detergents. In addition, npS-15615 performed well in the washing experiment, and the washing effect at 20 ℃ can be significantly improved by adding crude enzyme solution in the washing process.
Topics: Detergents; Hydrogen-Ion Concentration; Temperature; Metalloproteases; Recombinant Proteins; Bacterial Proteins; Bacillus licheniformis; Enzyme Stability; Planococcus Bacteria; Caseins; Gene Expression; Cloning, Molecular; Surface-Active Agents; Hydrolysis
PubMed: 37880082
DOI: 10.2323/jgam.2023.09.002 -
PloS One 2018The weak interlayer refers to the filling material in shear belts or large-scale structural planes, which is usually composed of soil, fine sand and gravels. It is prone...
The weak interlayer refers to the filling material in shear belts or large-scale structural planes, which is usually composed of soil, fine sand and gravels. It is prone to argillization when encountering water and its mechanical strength and stiffness are generally low, which has adverse effects on the stability of underground structures. In recent years, research on reinforcement techniques for weak interlayers has been a hot topic in geotechnical field. As a new reinforcement method for structural planes, the microbial healing technique has attracted a lot of attention. In this paper, a study on the healing technique for weak interlayer based on microbially induced calcium carbonate precipitation (MICP) and related mechanical properties was conducted for the interlayer shear belt at Baihetan Hydropower Station in China. First, Sporosarcina pasteurii was activated in laboratory. Reinforcement of the weak interlayer was realized by utilizing calcium carbonate precipitation on the weak interlayer. Continuous monitoring of the precipitates on the weak interlayer by XRD and SEM indicated that the precipitates on the weak layer were microbially induced calcium carbonate. Its crystals were irregular fish scale-shaped cubes with size in the range of 5~20μm. With favorable crystal growth, the crystals and the particles of the weak interlayer were cemented together. Finally, the mechanical properties of the healed weak interlayer were tested and the variations of uniaxial compressive strength, shear strength and triaxial compressive strength with bacteria concentration were discussed. The test results indicated that the maximum uniaxial compressive strength, peak shear strength and triaxial compressive strength can be increased by 149%, 162% and 119%, respectively, which subsequently improve the overall strength of the shear zone or structural plane. This can provide a new idea for soft ground reinforcement in underground projects.
Topics: Calcium Carbonate; Chemical Precipitation; Construction Industry; Microscopy, Electron, Scanning; Power Plants; Shear Strength; Soil Microbiology; Sporosarcina
PubMed: 30212556
DOI: 10.1371/journal.pone.0203834 -
Acta Crystallographica. Section F,... Nov 2018The determination of conditions for the reproducible growth of well diffracting crystals is a critical step in every biocrystallographic study. On the occasion of a new...
The determination of conditions for the reproducible growth of well diffracting crystals is a critical step in every biocrystallographic study. On the occasion of a new structural biology project, several advanced crystallogenesis approaches were tested in order to increase the success rate of crystallization. These methods included screening by microseed matrix screening, optimization by counter-diffusion and crystal detection by trace fluorescent labeling, and are easily accessible to any laboratory. Their combination proved to be particularly efficient in the case of the target, a 48 kDa CCA-adding enzyme from the psychrophilic bacterium Planococcus halocryophilus. A workflow summarizes the overall strategy, which led to the production of crystals that diffracted to better than 2 Å resolution and may be of general interest for a variety of applications.
Topics: Bacterial Proteins; Crystallization; Crystallography, X-Ray; Escherichia coli; Planococcus Bacteria; RNA Nucleotidyltransferases; Recombinant Proteins; Workflow
PubMed: 30387781
DOI: 10.1107/S2053230X18014590 -
Microbial Cell Factories Jun 2024Microbially induced calcium carbonate precipitation has been extensively researched for geoengineering applications as well as diverse uses within the built environment....
BACKGROUND
Microbially induced calcium carbonate precipitation has been extensively researched for geoengineering applications as well as diverse uses within the built environment. Bacteria play a crucial role in producing calcium carbonate minerals, via enzymes including carbonic anhydrase-an enzyme with the capability to hydrolyse CO, commonly employed in carbon capture systems. This study describes previously uncharacterised carbonic anhydrase enzyme sequences capable of sequestering CO2 and subsequentially generating CaCO biominerals and suggests a route to produce carbon negative cementitious materials for the construction industry.
RESULTS
Here, Bacillus subtilis was engineered to recombinantly express previously uncharacterised carbonic anhydrase enzymes from Bacillus megaterium and used as a whole cell catalyst allowing this novel bacterium to sequester CO and convert it to calcium carbonate. A significant decrease in CO was observed from 3800 PPM to 820 PPM upon induction of carbonic anhydrase and minerals recovered from these experiments were identified as calcite and vaterite using X-ray diffraction. Further experiments mixed the use of this enzyme (as a cell free extract) with Sporosarcina pasteurii to increase mineral production whilst maintaining a comparable level of CO sequestration.
CONCLUSION
Recombinantly produced carbonic anhydrase successfully sequestered CO and converted it into calcium carbonate minerals using an engineered microbial system. Through this approach, a process to manufacture cementitious materials with carbon sequestration ability could be developed.
Topics: Calcium Carbonate; Bacillus subtilis; Carbon Dioxide; Carbonic Anhydrases; Sporosarcina; Bacillus megaterium; Carbon Sequestration; Chemical Precipitation; Bacterial Proteins
PubMed: 38858761
DOI: 10.1186/s12934-024-02437-7 -
Food Microbiology Sep 2019The Minas artisanal cheese is a traditional product in its way of producing. Produced in the Minas Gerais state, Brazil, this cheese is made using raw cow's milk with...
The Minas artisanal cheese is a traditional product in its way of producing. Produced in the Minas Gerais state, Brazil, this cheese is made using raw cow's milk with the addition of an endogenous starter culture called "pingo", responsible for inoculating specific microorganisms that could enhance flavor and sensorial aspects. There are seven regions able to produce and commercialize this product - Araxá, Campo das Vertentes, Canastra Cerrado, Serra do Salitre, Serro and Triângulo Mineiro. This study aimed to assess the bacterial community of raw milk, endogenous starter culture and to uncover possible shifts in the bacterial community of the rind and core of cheeses at sixty days of ripening located in the Serra do Salitre region by Illumina MiSeq 16S rRNA gene amplicon sequencing. Raw milk and starter culture are responsible for inoculating specific bacteria into the cheese, with Planococcaceae and Streptococcaceae being prevalent throughout ripening time. The Planococcaceae family seems to develop strong interactions with the Leuconostocaceae family on the surface of these cheeses, and is associated with environmental aspects of the region, probably leading to a microbial signature of these products. Additionally, abiotic factors such as geographical location, moisture and acidity are major drivers in the microbial shift.
Topics: Animals; Bacteria; Brazil; Cheese; Environment; Food Microbiology; Microbiota; Milk; RNA, Ribosomal, 16S; Sensation; Taste; Time Factors
PubMed: 31027793
DOI: 10.1016/j.fm.2019.02.016 -
The ISME Journal Jun 2013Planococcus halocryophilus strain Or1, isolated from high Arctic permafrost, grows and divides at -15 °C, the lowest temperature demonstrated to date, and is...
Planococcus halocryophilus strain Or1, isolated from high Arctic permafrost, grows and divides at -15 °C, the lowest temperature demonstrated to date, and is metabolically active at -25 °C in frozen permafrost microcosms. To understand how P. halocryophilus Or1 remains active under the subzero and osmotically dynamic conditions that characterize its native permafrost habitat, we investigated the genome, cell physiology and transcriptomes of growth at -15 °C and 18% NaCl compared with optimal (25 °C) temperatures. Subzero growth coincides with unusual cell envelope features of encrustations surrounding cells, while the cytoplasmic membrane is significantly remodeled favouring a higher ratio of saturated to branched fatty acids. Analyses of the 3.4 Mbp genome revealed that a suite of cold and osmotic-specific adaptive mechanisms are present as well as an amino acid distribution favouring increased flexibility of proteins. Genomic redundancy within 17% of the genome could enable P. halocryophilus Or1 to exploit isozyme exchange to maintain growth under stress, including multiple copies of osmolyte uptake genes (Opu and Pro genes). Isozyme exchange was observed between the transcriptome data sets, with selective upregulation of multi-copy genes involved in cell division, fatty acid synthesis, solute binding, oxidative stress response and transcriptional regulation. The combination of protein flexibility, resource efficiency, genomic plasticity and synergistic adaptation likely compensate against osmotic and cold stresses. These results suggest that non-spore forming P. halocryophilus Or1 is specifically suited for active growth in its Arctic permafrost habitat (ambient temp. ∼-16 °C), indicating that such cryoenvironments harbor a more active microbial ecosystem than previously thought.
Topics: Acclimatization; Arctic Regions; Cell Membrane; Cold Temperature; Ecosystem; Freezing; Gene Expression Regulation, Bacterial; Genome, Bacterial; Molecular Sequence Data; Osmotic Pressure; Planococcus Bacteria
PubMed: 23389107
DOI: 10.1038/ismej.2013.8