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Current Research in Microbial Sciences Dec 2021The proteomics and transcriptomic analysis of strain AS2 was carried out through 2D gel electrophoresis and RT-PCR. Seven protein spots were selected randomly from the...
The proteomics and transcriptomic analysis of strain AS2 was carried out through 2D gel electrophoresis and RT-PCR. Seven protein spots were selected randomly from the gel and identified through mass spectrometry. Four proteins including putative metal-dependent hydrolase TatD, thioredoxin reductase, DNA-directed RNA polymerase subunit alpha and chaperone protein DnaK were upregulated while superoxide dismutase [Mn], 3-oxoacyl-[acyl-carrier-protein] reductase FabG, and putative alkyl/aryl-sulfatase YjcS were down-regulated under arsenite stress. No significant difference was observed in gene expression analysis in the presence and absence of arsenite. The optimum arsenite processing ability was determined at 37°C (90%) and at pH 7 (92%). The maximum metal processing ability was determined at 250 mM arsenite/L (90%) while the minimum was estimated at 1250 mM arsenite/L (42%). The maximum arsenite removal ability of strain AS2 determined after 8 days was 68 and 82% from wastewater and distilled water, and the organism can be a good bioresource for green chemistry to eradicate environmental arsenite.
PubMed: 34841312
DOI: 10.1016/j.crmicr.2021.100020 -
Scientific Reports Nov 2022This study investigated the mechanism of membrane damage by protocatechualdehyde (PCA) against Micrococcus luteus and assessed effects of PCA on the sensory and...
This study investigated the mechanism of membrane damage by protocatechualdehyde (PCA) against Micrococcus luteus and assessed effects of PCA on the sensory and physicochemical properties of pork. The mechanism of PCA inhibition on M. luteus was studied by determining the minimum inhibitory concentration (MIC) based on membrane potential, intracellular ATP concentration, intracellular pH, confocal laser scanning microscopy (CLSM), and field emission gun scanning electron microscopy (FEG-SEM). The results showed that the MIC of PCA against M. luteus was 1.25 mg/mL. Hyperpolarization of the bacterial cell membrane, a decrease in the intracellular ATP concentration, and intracellular pH indicated that PCA damaged the cell membrane of M. luteus. FEG-SEM observation revealed that PCA could cause surface collapse, cell membrane rupture, and content outflow of M. luteus. Additionally, PCA was found to inhibit increases in the total number of colonies, the thiobarbituric acid reactive substances (TBARS) value growth rate, and moisture mobility in raw pork. Additionally, it improved the color and texture of raw pork, all of which effectively prolonged its shelf life. This study will encourage the application of PCA as a natural antibacterial agent in the food industry.
Topics: Animals; Swine; Micrococcus luteus; Pork Meat; Red Meat; Adenosine Triphosphate
PubMed: 36344587
DOI: 10.1038/s41598-022-23309-3 -
BMC Genomics Feb 2021Micrococcus luteus is a group of actinobacteria that is widely used in biotechnology and is being thought as an emerging nosocomial pathogen. With one of the smallest...
BACKGROUND
Micrococcus luteus is a group of actinobacteria that is widely used in biotechnology and is being thought as an emerging nosocomial pathogen. With one of the smallest genomes of free-living actinobacteria, it is found in a wide range of environments, but intraspecies genetic diversity and adaptation strategies to various environments remain unclear. Here, comparative genomics, phylogenomics, and genome-wide association studies were used to investigate the genomic diversity, evolutionary history, and the potential ecological differentiation of the species.
RESULTS
High-quality genomes of 66 M. luteus strains were downloaded from the NCBI GenBank database and core and pan-genome analysis revealed a considerable intraspecies heterogeneity. Phylogenomic analysis, gene content comparison, and average nucleotide identity calculation consistently indicated that the species has diverged into three well-differentiated clades. Population structure analysis further suggested the existence of an unknown ancestor or the fourth, yet unsampled, clade. Reconstruction of gene gain/loss events along the evolutionary history revealed both early events that contributed to the inter-clade divergence and recent events leading to the intra-clade diversity. We also found convincing evidence that recombination has played a key role in the evolutionary process of the species, with upto two-thirds of the core genes having been affected by recombination. Furthermore, distribution of mammal-associated strains (including pathogens) on the phylogenetic tree suggested that the last common ancestor had a free-living lifestyle, and a few recently diverged lineages have developed a mammal-associated lifestyle separately. Consistently, genome-wide association analysis revealed that mammal-associated strains from different lineages shared genes functionally relevant to the host-associated lifestyle, indicating a recent ecological adaption to the new host-associated habitats.
CONCLUSIONS
These results revealed high intraspecies genomic diversity of M. luteus and highlighted that gene gain/loss events and extensive recombination events played key roles in the genome evolution. Our study also indicated that, as a free-living species, some lineages have recently developed or are developing a mammal-associated lifestyle. This study provides insights into the mechanisms that drive the genome evolution and adaption to various environments of a bacterial species.
Topics: Animals; Evolution, Molecular; Genetic Variation; Genome, Bacterial; Genome-Wide Association Study; Genomics; Micrococcus luteus; Phylogeny; Recombination, Genetic
PubMed: 33602135
DOI: 10.1186/s12864-021-07432-5 -
3 Biotech Jul 2021In this study, hydrolytic and oxidative activities of enzymes isolated from halophilic microbes were characterized and applied for biomass utilization. First, lipase...
In this study, hydrolytic and oxidative activities of enzymes isolated from halophilic microbes were characterized and applied for biomass utilization. First, lipase from , and peroxidase and laccase from and were selected and their catalytic activities were determined, respectively. The lipase encoding gene was synthesized after codon-optimization and could be successfully expressed in with the assist of the Tif chaperone protein. The purified enzyme showed 119.13 ± 7.18 and 34.42 ± 5.91 U/mL of lipase and esterase activities, respectively. Moreover, the lipase was applied for hydrolysis of the triglycerides mixture, which resulted in 182.9 ± 11.1 mg/L/h of glycerol productivity. Next, peroxidase and laccase activities of and were determined, and extracellular enzymes of was applied for lignocellulosic biomass degradation, which resulted in 91.9 μg glucose/mg lignocellulose of production yields. Finally, the hydrolytic and oxidative activities of the enzymes from halophilic microbes could be further utilized for biomass treatment and biochemical production.
PubMed: 34295605
DOI: 10.1007/s13205-021-02902-9 -
Frontiers in Pediatrics 2023To explore the clinical characteristics of bloodstream infection in an infant and characterize the phenotype and genotype of the isolated strains, as well as seek...
PURPOSE
To explore the clinical characteristics of bloodstream infection in an infant and characterize the phenotype and genotype of the isolated strains, as well as seek suitable infection models for assessing virulence.
METHODS
Clinical data was collected from an infant patient diagnosed with bloodstream infection. Metagenomic sequencing was performed on the isolated blood sample. The strain was isolated and underwent mass spectrometry, biochemical tests, antibiotic susceptibility assays, and whole-genome sequencing. The infection model was used to assess virulence.
RESULTS
Patient responded poorly to cephalosporins, but recovered after Linezolid treatment. Metagenomic sequencing identified as the predominant species in the sample, confirming infection. They were identified as with a high confidence level of 98.99% using mass spectrometry. The strain showed positive results for Catalase, Oxidase, and Urea tests, and negative results for Mannose, Xylose, Lactose, Mannitol, Arginine, and Galactose tests, consistent with the biochemical profile of reference standards. susceptibility to 15 antibiotics was demonstrated and no resistance genes were detected. Predicted virulence genes, including , were associated with metabolic pathways and the type VI secretion system. The infection model demonstrated dose-dependent survival rates.
CONCLUSION
The infant likely developed a bloodstream infection with due to compromised immunity. Although the isolated strain is sensitive to cephalosporin antibiotics and has low pathogenicity in infection models, clinical treatment with cephalosporins was ineffective. Linezolid proved to be effective, providing valuable guidance for future clinical management of such rare infections.
PubMed: 38188910
DOI: 10.3389/fped.2023.1303040 -
Plants (Basel, Switzerland) Aug 2022Cadmium (Cd) and nickel (Ni) are two of the most toxic metals, wreaking havoc on human health and agricultural output. Furthermore, high levels of Cd and Ni in the soil...
Cadmium (Cd) and nickel (Ni) are two of the most toxic metals, wreaking havoc on human health and agricultural output. Furthermore, high levels of Cd and Ni in the soil environment, particularly in the root zone, may slow plant development, resulting in lower plant biomass. On the other hand, endophytic bacteria offer great promise for reducing Cd and Ni. Moreover, they boost plants' resistance to heavy metal stress. Different bacterium strains were isolated from tomato roots. These isolates were identified as and using 16SrDNA and were utilized to investigate their involvement in mitigating the detrimental effects of heavy metal stress. The two bacterial strains can solubilize phosphorus and create phytohormones as well as siderophores. Therefore, the objective of this study was to see how endophytic bacteria ( and ) affected the mitigation of stress from Cd and Ni in tomato plants grown in 50 μM Cd or Ni-contaminated soil. According to the findings, Cd and Ni considerably lowered growth, biomass, chlorophyll (Chl) content, and photosynthetic properties. Furthermore, the content of proline, phenol, malondialdehyde (MDA), HO, OH, O, the antioxidant defense system, and heavy metal (HM) contents were significantly raised under HM-stress conditions. However, endophytic bacteria greatly improved the resistance of tomato plants to HM stress by boosting enzymatic antioxidant defenses (i.e., catalase, peroxidase, superoxide dismutase, glutathione reductase, ascorbate peroxidase, lipoxygenase activity, and nitrate reductase), antioxidant, non-enzymatic defenses, and osmolyte substances such as proline, mineral content, and specific regulatory defense genes. Moreover, the plants treated had a higher value for bioconcentration factor (BCF) and translocation factor (TF) due to more extensive loss of Cd and Ni content from the soil. To summarize, the promotion of endophytic bacterium-induced HM resistance in tomato plants is essentially dependent on the influence of endophytic bacteria on antioxidant capacity and osmoregulation.
PubMed: 35956496
DOI: 10.3390/plants11152018 -
Tropical Biomedicine Mar 2018The multifarious types of infections contracted from indoor environments show that buildings can serve as a reservoir for infectious bacteria. This study is an...
The multifarious types of infections contracted from indoor environments show that buildings can serve as a reservoir for infectious bacteria. This study is an investigation into the type and concentrations of bacteria in the indoor and outdoor environments of an electronic factory, an office and a winery in Malaysia. Trypticase soy agar (TSA) (with ambient air incubation) and TSA supplemented with haemin and NADH (with CO enhanced incubation) were used for the isolation of bacteria. The plates were incubated at 37ºC for 3 days. A random selection of bacterial isolates were Gram stained and identified using the BD BBL Crystal Identification Systems. Kytococcus sedentarius and Micrococcus luteus were the predominant bacterial species identified from indoor air. These bacteria were present at relatively high concentrations in indoor air, at times, above 800 colony forming units per cubic meter (CFU/m) of air. This indicates that both K. sedentarius and M. luteus can survive a wide range of adverse conditions, including chemical contamination and ultraviolet exposure. M. luteus is a known cause of pneumonia in immunocompromised individuals and has also been implicated in skin infections. Recent reports suggest species of kytococci as emerging opportunistic pathogens of the immunocompromised, paediatrics and the elderly. We postulate that opportunistic bacteria, such as the kytococci and the micrococci, may also have a potential role in instigating subclinical, more subtle symptoms of disease in inmmunocompetent individuals.
PubMed: 33601787
DOI: No ID Found -
Access Microbiology 2023Dormant bacterial cells do not divide and are not immediately culturable, but they persist in a state of low metabolic activity, a physiological state having clinical...
Dormant bacterial cells do not divide and are not immediately culturable, but they persist in a state of low metabolic activity, a physiological state having clinical relevance, for instance in latent tuberculosis. Resuscitation-promoting factors (Rpfs) are proteins that act as signalling molecules mediating growth and replication. In this study we aimed to test the effect of Rpfs from on the number and diversity of cultured bacteria using insect and soil samples, and to examine if the increase in culturability could be reproduced with the putative reaction product of Rpf, 1,6-anhydro--acetylmuramic acid (1,6-anhydro-MurNAc). The gene from was amplified and cloned into a pET21b expression vector and the protein was expressed in BL21(DE3) cells and purified by affinity chromatography using a hexa-histidine tag. 1,6-Anhydro-MurNAc was prepared using reported chemical synthesis methods. Recombinant Rpf protein or 1,6-anhydro-MurNAc were added to R2A cultivation media, and their effect on the culturability of bacteria from eight environmental samples including four cockroach guts and four soils was examined. Colony-forming units, 16S rRNA gene copies and Illumina amplicon sequencing of the 16S rRNA gene were measured for all eight samples subjected to three different treatments: Rpf, 1,6-anhydro-MurNAc or blank control. Both Rpf and 1,6-anhydro-MurNAc increased the number of colony-forming units and of 16S rRNA gene copies across the samples although the protein was more effective. The Rpf and 1,6-anhydro-MurNAc promoted the cultivation of a diverse set of bacteria and in particular certain clades of the phyla and . This study opens the path for improved cultivation strategies aiming to isolate and study yet undescribed living bacterial organisms.
PubMed: 37841103
DOI: 10.1099/acmi.0.000647.v4 -
Pharmaceutics Jun 2021Infectious diseases caused by microbial biofilms are a major clinical problem, and new antimicrobial agents that can inhibit biofilm formation and eradicate pre-formed...
Infectious diseases caused by microbial biofilms are a major clinical problem, and new antimicrobial agents that can inhibit biofilm formation and eradicate pre-formed biofilms are urgently needed. Pomegranate extracts are a well-established folkloric medicine and have been used in the treatment of infectious diseases since ancient times, whilst the addition of metal ions, including zinc (II), has enhanced the antimicrobial activity of pomegranate. is generally a non-pathogenic skin commensal bacterium, although it can act as an opportunistic pathogen and cause serious infections, particularly involving catheterization and comorbidities. The aims of this study were to evaluate the holistic activity of pomegranate rind extract (PRE), Zn (II), and PRE/Zn (II) individually and in combination against under both planktonic and biofilm conditions. Antimicrobial activity was detected in vitro using the broth dilution method, and synergistic activity was determined using checkerboard and time-kill assays. Effects on biofilm formation and eradication were determined by crystal violet and BacLight Live/Dead staining. PRE and Zn (II) exerted antimicrobial activity against under both planktonic and biofilm conditions. After 4 h, potent synergistic bactericidal activity was also found when PRE and Zn (II) were co-administered under planktonic conditions (log reductions: PRE 1.83 ± 0.24, Zn (II) 3.4 ± 0.08, and PRE/Zn (II) 6.88 ± 1.02; < 0.0001). In addition, greater heterogeneity was induced in the structure of biofilm using the PRE/Zn (II) combination compared to when PRE and Zn (II) were applied individually. The activity of PRE and the PRE/Zn (II) combination could offer a novel antimicrobial therapy for the treatment of disease-associated infections caused by and potentially other bacteria.
PubMed: 34201223
DOI: 10.3390/pharmaceutics13060851 -
Journal, Genetic Engineering &... Apr 2021Lipases are promising biocatalysts for industrial applications and attract attention to be explored. A novel acidic lipase has been isolated from the lipolytic bacteria...
BACKGROUND
Lipases are promising biocatalysts for industrial applications and attract attention to be explored. A novel acidic lipase has been isolated from the lipolytic bacteria Micrococcus luteus EMP48-D (LipEMP48-D) screened from tempeh. The lipase gene had previously been overexpressed in Escherichia coli BL21, but the expression level obtained was relatively low. Here, to improve the expression level, the lipase gene was cloned to Pichia pastoris. We eliminated the native signal sequence of M. luteus and replaced it with α-mating factor (α-MF) signal sequence. We also optimized and synthesized the lipase gene based on codon preference in P. pastoris.
RESULTS
LipEMP48-D lipase was expressed as an extracellular protein. Codon optimization has been conducted for 20 codons, with the codon adaption index reaching 0.995. The highest extracellular lipase activity obtained reached 145.4 ± 4.8 U/mg under AOX1 promoter in P. pastoris KM71 strain, which was 9.7-fold higher than the previous activity in E. coli. LipEMP48-D showed the highest specific activity at pH 5.0 and stable within the pH range 3.0-5.0 at 40 °C. LipEMP48-D also has the capability of hydrolyzing various long-chain triglycerides, particularly olive oil (100%) followed by sunflower oil (88.5%). LipEMP48-D exhibited high tolerance for various polar organic solvents with low log P, such as isopropanol (115.7%) and butanol (114.6%). The metal ions (Na, K, Ca, Mg, Mn) decreased enzyme activity up to 43.1%, while Fe increased relative activity of enzymes up to 200%. The conversion of free fatty acid (FFA) into fatty acid methyl ester (FAME) was low around 2.95%.
CONCLUSIONS
This study was the first to report overexpression of Micrococcus lipase in yeast. The extracellular expression of this acidic lipase could be potential for biocatalyst in industrial fields, especially organic synthesis, food industry, and production of biodiesel.
PubMed: 33826047
DOI: 10.1186/s43141-021-00155-w