Bacillus strain NS 8, a lipase-producing bacterium isolated from a Malaysian hot spring, is able to tolerate a broad range of temperature and pH, which makes it beneficial for this study. It generated PCR products with molecular weight of 1,532 bp, and the 16S rRNA sequence analysis identified it as Bacillus subtilis with accession number AB110598. It showed a 71% similarity index with B. subtilis using Biolog Microstation System. Its lipase production was optimized using a shake flask system by changing the physical (agitation speed, pH and temperature) and nutritional (nitrogen, carbon and minerals) factors. The most suitable combination of the basal medium for lipase production was 2.5% olive oil (carbon), 1.5% peptone (nitrogen), 0.1% MgSO(4) (mineral) at an optimum temperature of 50°C, pH 7.5 and 150 rpm agitation, giving an enzyme yield of 4.23 U/ml. Statistical optimization using response surface methodology was carried out. An optimum lipase production of 5.67 U/ml was achieved when olive oil concentration of 3%, peptone 2%, MgSO(4)·7H(2)O 0.2% and an agitation rate of 200 rpm were combined. Lipase production was further carried out inside a 2-liter bioreactor, which yielded an enzyme activity of 14.5 U/ml after 15 h of incubation.
Thermostable lipase produced by a genotypically identified extremophilic Bacillus subtilis NS 8 was purified 500-fold to homogeneity with a recovery of 16% by ultrafiltration, DEAE-Toyopearl 650M and Sephadex G-75 column. The purified enzyme showed a prominent single band with a molecular weight of 45 kDa. The optimum pH and temperature for activity of lipase were 7.0 and 60°C, respectively. The enzyme was stable in the pH range between 7.0 and 9.0 and temperature range between 40 and 70°C. It showed high stability with half-lives of 273.38 min at 60°C, 51.04 min at 70°C and 41.58 min at 80°C. The D-values at 60, 70 and 80°C were 788.70, 169.59 and 138.15 min, respectively. The enzyme's enthalpy, entropy and Gibb's free energy were in the range of 70.07-70.40 kJ mol(-1), -83.58 to -77.32 kJ mol(-1)K(-1) and 95.60-98.96 kJ mol(-1), respectively. Lipase activity was slightly enhanced when treated with Mg(2+) but there was no significant enhancement or inhibition of the activity with Ca(2+). However, other metal ions markedly inhibited its activity. Of all the natural vegetable oils tested, it had slightly higher hydrolytic activity on soybean oil compared to other oils. On TLC plate, the enzyme showed non-regioselective activity for triolein hydrolysis.
Matched MeSH terms: Bacillus subtilis/enzymology*; Bacillus subtilis/growth & development
Actinopyga lecanora, a type of sea cucumber commonly known as stone fish with relatively high protein content, was explored as raw material for bioactive peptides production. Six proteolytic enzymes, namely alcalase, papain, pepsin, trypsin, bromelain and flavourzyme were used to hydrolyze A. lecanora at different times and their respective degrees of hydrolysis (DH) were calculated. Subsequently, antibacterial activity of the A. lecanora hydrolysates, against some common pathogenic Gram positive bacteria (Bacillus subtilis and Staphylococcus aureus) and Gram negative bacteria (Escherichia coli, Pseudomonas aeruginosa, and Pseudomonas sp.) were evaluated. Papain hydrolysis showed the highest DH value (89.44%), followed by alcalase hydrolysis (83.35%). Bromelain hydrolysate after one and seven hours of hydrolysis exhibited the highest antibacterial activities against Pseudomonas sp., P. aeruginosa and E. coli at 51.85%, 30.07% and 30.45%, respectively compared to the other hydrolysates. Protein hydrolysate generated by papain after 8 h hydrolysis showed maximum antibacterial activity against S. aureus at 20.19%. The potent hydrolysates were further fractionated using RP-HPLC and antibacterial activity of the collected fractions from each hydrolysate were evaluated, wherein among them only three fractions from the bromelain hydrolysates exhibited inhibitory activities against Pseudomonas sp., P. aeruginosa and E. coli at 24%, 25.5% and 27.1%, respectively and one fraction of papain hydrolysate showed antibacterial activity of 33.1% against S. aureus. The evaluation of the relationship between DH and antibacterial activities of papain and bromelain hydrolysates revealed a meaningful correlation of four and six order functions.
In this study, potential probiotic strains were isolated from fermented pickles based on antagonistic activity against two shrimp pathogens (Vibrio harveyi and Vibrio parahaemolyticus). Two strains L10 and G1 were identified by biochemical tests, followed by16S ribosomal RNA gene sequence analysis as Bacillus subtilis, and characterized by PCR amplification of repetitive bacterial DNA elements (Rep-PCR). Subsequently, B. subtilis L10 and G1 strains were tested for antibacterial activity under different physical conditions, including culture medium, salinity, pH and temperature using the agar well diffusion assay. Among the different culture media, LB broth was the most suitable medium for antibacterial production. Both strains showed the highest level of antibacterial activity against two pathogens at 30 °C and 1.0% NaCl. Under the pH conditions, strain G1 showed the greatest activity against V. harveyi at pH 7.3-8.0 and against V. parahaemolyticus at pH 6.0-8.0, whereas strain L10 showed the greatest activity against two pathogens at pH 7.3. The cell-free supernatants of both strains were treated with four different enzymes in order to characterize the antibacterial substances against V. harveyi. The result showed considerable reduction of antibacterial activity for both strains, indicating the proteinaceous nature of the antibacterial substances. A wide range of tolerance to NaCl, pH and temperature was also recorded for both strains. In addition, both strains showed no virulence effect in juvenile shrimp Litopenaeus vannamei. On the basis of these results and safety of strains to L. vannamei, they may be considered for future challenge experiments in shrimp as a very promising alternative to the use of antibiotics.
Antimicrobial irradiated seaweed-neem biocomposite films were synthesized in this study. The storage functional properties of the films were investigated. Characterization of the prepared films was conducted using SEM, FT-IR, contact angle, and antimicrobial test. The macroscopic and microscopic including the analysis of the functional group and the gas chromatography-mass spectrometry test revealed the main active constituents present in the neem extract, which was used an essential component of the fabricated films. Neem leaves' extracts with 5% w/w concentration were incorporated into the matrix of seaweed biopolymer and the seaweed-neem bio-composite film were irradiated with different dosages of gamma radiation (0.5, 1, 1.5, and 2 kGy). The tensile, thermal, and the antimicrobial properties of the films were studied. The results revealed that the irradiated films exhibited improved functional properties compared to the control film at 1.5 kGy radiation dosage. The tensile strength, tensile modulus, and toughness exhibited by the films increased, while the elongation of the irradiated bio-composite film decreased compared to the control film. The morphology of the irradiated films demonstrated a smoother surface compared to the control and provided surface intermolecular interaction of the neem-seaweed matrix. The film indicated an optimum storage stability under ambient conditions and demonstrated no significant changes in the visual appearance. However, an increase in the moisture content was exhibited by the film, and the hydrophobic properties was retained until nine months of the storage period. The study of the films antimicrobial activities against Staphylococcus aureus (SA), and Bacillus subtilis (BS) indicated improved resistance to bacterial activities after the incorporation of neem leaves extract and gamma irradiation. The fabricated irradiated seaweed-neem bio-composite film could be used as an excellent sustainable packaging material due to its effective storage stability.
The bioactivity of R. nasutus leaf extracts was assessed on Bacillus cereus, Bacillus subtilis, Staphylococcus aureus, Streptococcus pyogenes, Vibrio parahaemolyticus, Enterobacter aerogenes, Proteus mirabilis, and Klebsiella pneumoniae. Crude chloroform, petroleum ether, ethyl acetate, ethanol and methanol extracts were screened by disc diffusion method. Promising crude extract was further subjected to the column fractionation followed by the screening of the antibacterial activity of individual fractions. Biologically active pure fraction was subjected to the advanced analytical studies like HPLC, LC-MS, IR and NMR for characterisation of the bioactive compound. Ethanolic extract exhibited the maximum antibacterial activity against Klebsiella pneumoniae with the maximum of 35±0.42 mm zone of inhibition. The biologically potent column fraction from ethanol extract with 40±0.42 mm zone of inhibition upon subject to the HPLC, LC-MS, IR and NMR revealed that the active compound is rhinacanthin-C, a naphthoquinone.
Disinfectants are generally used to inactivate microorganisms in solutions. The process of inactivation involves the disinfectant in the liquid diffusing towards the bacteria sites and thereafter reacting with bacteria at rates determined by the respective reaction rates. Such processes have demonstrated an initial lag phase followed by an active depletion phase of bacteria. This paper attempts to study the importance of the combined effects of diffusion of the disinfectant through the outer membrane of the bacteria and transport through the associated concentration boundary layers (CBLs) during the initial lag phase. Mathematical equations are developed correlating the initial concentration of the disinfectant with time required for reaching a critical concentration (C*) at the inner side of the membrane of the cell based on diffusion of disinfectant through the outer membranes of the bacteria and the formation of concentration boundary layers on both sides of the membranes. Experimental data of the lag phases of inactivation already available in the literature for inactivation of Bacillus subtilis spores with ozone and monochloramine are tested with the equations. The results seem to be in good agreement with the theoretical equations indicating the importance of diffusion process across the outer cell membranes and the resulting CBL's during the lag phase of disinfection.
While bulk zinc oxide (ZnO) is of non-toxic in nature, ZnO nanoarchitectures could potentially induce the macroscopic characteristics of oxidative, lethality and toxicity in the water environment. Here we report a systematic study through state-of-the-art controllable synthesis of multi-dimensional ZnO nanoarchitectures (i.e. 0D-nanoparticle, 1D-nanorod, 2D-nanosheet, and 3D-nanoflowers), and subsequent in-depth understanding on the fundamental factor that determines their photoactivities. The photoactivities of resultant ZnO nanoarchitectures were interpreted in terms of the photodegradation of salicylic acid as well as inactivation of Bacillus subtilis and Escherichia coli under UV-A irradiation. Photodegradation results showed that 1D-ZnO nanorods demonstrated the highest salicylic acid photodegradation efficiency (99.4%) with a rate constant of 0.0364 min-1. 1D-ZnO nanorods also exhibited the highest log reductions of B. subtilis and E. coli of 3.5 and 4.2, respectively. Through physicochemical properties standardisation, an intermittent higher k value for pore diameter (0.00097 min-1 per mm), the highest k values for crystallite size (0.00171 min-1 per nm) and specific surface area (0.00339 min-1 per m2/g) contributed to the exceptional photodegradation performance of nanorods. Whereas, the average normalised log reduction against the physicochemical properties of nanorods (i.e. low crystallite size, high specific surface area and pore diameter) caused the strongest bactericidal effect.
Matched MeSH terms: Bacillus subtilis/growth & development
We studied the effect of two probiotic Bacillus subtilis strains on the growth performance, digestive enzyme activity, immune gene expression and disease resistance of juvenile white shrimp (Litopenaeus vannamei). A mixture of two probiotic strains, L10 and G1 in equal proportions, was administered at two different doses 10(5) (BM5) and 10(8) (BM8) CFU g(-1) feed to shrimp for eight weeks. In comparison to untreated control group, final weight, weight gain and digestive enzyme activity were significantly greater in shrimp fed BM5 and BM8 diets. Significant differences for specific growth rate (SGR) and survival were recorded in shrimp fed BM8 diet as compared with the control; however, no significant differences were recorded for food conversion ratio (FCR) among all the experimental groups. Eight weeks after the start of the feeding period, shrimp were challenged with Vibrio harveyi. Statistical analysis revealed significant differences in shrimp survival between probiotic and control groups. Cumulative mortality of the control group was 63.3%, whereas cumulative mortality of the shrimp that had been given probiotics was 20.0% with BM8 and 33.3% with BM5. Subsequently, real-time PCR was employed to determine the mRNA levels of prophenoloxidase (proPO), peroxinectin (PE), lipopolysaccharide- and β-1,3-glucan-binding protein (LGBP) and serine protein (SP). The expression of all immune-related genes studied was significantly up-regulated (P
Endophytic fungi provide protection to their host plant and the fungi often produce antimicrobial compounds to aid the host
fighting off pathogens. These bioactive compounds were secondary metabolites which were often produced as waste- or
by-products. In the present study, endophytic fungi isolated from mangrove plants and soils were characterized and their
antimicrobial production and bioremediation potential of heavy metals copper (Cu) and zinc (Zn) were assessed. Twelve
(12) isolated and identified endophytic fungi belonged to seven species; Penicillium, Curvularia, Diaporthe, Aspergillus,
Guignardia, Neusartorya and Eupenicillium. Antimicrobial activities of these 12 fungal endophytes were tested against
Gram negative bacteria; Bacillus subtilis, Staphylococcus aureus, Gram positive bacteria; Escherichia coli and fungi;
Candida albicans and Aspergillus niger among others. Two isolates (related to Guignardia sp. and Neusartoya sp.) showed
strong antimicrobial (and antifungal) activity whereas the rest showed no activity. Compounds were isolated from both
isolates and screened using HPLC. Both isolates displayed chemically very interesting chromatograms as they possessed a
high diversity of basic chemical structures and peaks over a wide range of polarities, with structures similar to Trimeric
catechin and Helenalin among others. For bioremediation assessment, the results showed maximum biosorption capacity
for two isolates related to Curvularia sp. and Neusartorya sp., with the former removing 25 mg Cu/g biomass and the
latter removing 24 mg Zn/g biomass. Our results indicated the potential of mangrove endophytic fungi in producing
bioactive compounds and also highlighted their potential for the treatment of heavy metal-contaminated wastewater.
A new dichlorinated pulvinic acid derivative, methyl-3',5'-dichloro-4,4'-di-O-methylatromentate, was isolated from the fruiting body of a Scleroderma sp. The structure was determined using spectroscopic methods, and an X-ray analysis was carried out for confirmation of the structure. Compound was found to display moderate antimicrobial activity against Bacillus subtilis.
Cupressus sempervirens is a known traditional plant used to manage various ailments, including cancer, inflammatory and infectious diseases. In this investigation, we aimed to explore the chemical profile of Cupressus sempervirens essential oil (CSEO) as well as their antibacterial mode of action. The volatile components were characterized using gas chromatography coupled to a mass spectrometer (GC-MS). The results revealed remarkable antibacterial properties of EO derived from C. sempervirens. GC-MS analysis indicated that C. sempervirens EO characterized by δ-3-carene (47.72%), D-limonene (5.44%), β-pinene (4.36%), β-myrcene (4.02%). The oil exhibited significant inhibitory effects against a range of bacteria, including Staphylococcus aureus ATCC 29213, Bacillus subtilis ATCC 13048, Bacillus cereus (Clinical isolate), Pseudomonas aeruginosa ATCC 27853, and Escherichia coli ATCC 25922. These inhibitory effects surpassed those of conventional antibiotics. Furthermore, the EO demonstrated low minimum inhibitory concentrations (MICs) and minimum bactericidal concentrations (MBCs), indicating its bactericidal nature (MBC/MIC < 4.0). Time-kill kinetics analysis showed that CSEO was particularly effective at 2 × MIC doses, rapidly reduced viable count of B. subtilis and P. aeruginosa within 8 h. This suggests that the oil acts quickly and efficiently. The cell membrane permeability test further demonstrated the impact of CSEO on the relative conductivity of B. subtilis and P. aeruginosa, both at 2 × MIC concentrations. These observations suggest that EO disrupts the bacterial membrane, thereby influencing their growth and viability. Additionally, the cell membrane integrity test indicated that the addition of CSEO to bacterial cultures resulted in the significant release of proteins from the bacterial cells. This suggests that EO affects the structural integrity of the bacterial cells. Furthermore, the anti-biofilm assay confirmed the efficacy of CSEO as a potent anti-biofilm agent. It demonstrated the oil's ability to inhibit quorum sensing, a crucial mechanism for biofilm formation, and its competitive performance compared to the tested antibiotics.
Biodegradation of pharmaceuticals active compounds (PACs) in secondary effluents by using B. subtilis 2012WTNC as a function of β-lactamase was optimized using response surface methodology (RSM) designed by central composite design (CCD). Four factors including initial concentration of bacteria (1-6 log10 CFU mL-1), incubation period (1-14 days), incubation temperature (20-40 °C) and initial concentration of PACs (1-5 mg L-1) were investigated. The optimal operating factors for biodegradation process determined using response surface methodology (RSM) was recorded with 5.57 log10 CFU mL-1 of B. subtilis, for 10.38 days, at 36.62 °C and with 4.14 mg L-1 of (cephalexin/amoxicillin) with R2 coefficient of 0.99. The biodegradation was 83.81 and 93.94% respectively. The relationship among the independent variables was significant (p
Lansium domesticum Corr. is a fruit tree of the Meliaceae family, which is commonly found in SouthEast Asia with a wide range of varieties. This study investigated three varieties of L. domesticum; Duku, Langsat and Dokong for the phytochemical screening and antimicrobial activity. Seeds from the matured fruits were extracted using hexane, methanol and water. The crude extracts were screened for antimicrobial activities toward three bacteria, namely, Pseudomonas aeruginosa, Bacillus subtilis, and Staphylococcus aureus. The findings showed that Langsat seed extracts contained more groups of compounds compared with the other two varieties, and its methanol extract demonstrated the highest inhibition zones against the three bacteria. The crude methanol extract of Duku seeds showed inhibition zones only towards Bacillus subtilis at a high concentration (1.0 mgL-1), whilst the seed extracts of Dokong showed no inhibition zones towards any of the tested bacteria.
The draft genome here presents the sequence of Bacillus subtilis UMX-103. The bacterial strain was isolated from hydrocarbon-contaminated soil from Terengganu, Malaysia. The whole genome of the bacterium was sequenced using Illumina HiSeq 2000 sequencing platform. The genome was assembled using de novo approach. The genome size of UMX-103 is 4,234,627 bp with 4399 genes comprising 4301 protein-coding genes and 98 RNA genes. The analysis of assembled genes revealed the presence of 25 genes involved in biosurfactant production, where 14 of the genes are related to biosynthesis and 11 of the genes are in the regulation of biosurfactant productions. This draft genome will provide insights into the genetic bases of its biosurfactant-producing capabilities.
The goal of this study was to determine inhibitory effect of palm kernel expeller (PKE) peptides of different degree of hydrolysis (DH %) against spore-forming bacteria Bacillus cereus, Bacillus circulans, Bacillus coagulans, Bacillus licheniformis, Bacillus megaterium, Bacillus pumilus, Bacillus stearothermophillus, Bacillus subtilis, Bacillus thuringiensis, Clostridium perfringens; and non-spore-forming bacteria Escherichia coli, Lisinibacillus sphaericus, Listeria monocytogenes, Pseudomonas aeruginosa, Salmonella Typhimurium and Staphylococcus aureus.
In the present study, binary oxide (cadmium oxide [CdO])x (zinc oxide [ZnO])1-x nanoparticles (NPs) at different concentrations of precursor in calcination temperature were prepared using thermal treatment technique. Cadmium and zinc nitrates (source of cadmium and zinc) with polyvinylpyrrolidone (capping agent) have been used to prepare (CdO)x (ZnO)1-x NPs samples. The sample was characterized by X-ray diffraction (XRD), scanning electron microscopy, energy-dispersive X-ray (EDX), transmission electron microscopy (TEM), and Fourier transform infrared (FTIR) spectroscopy. XRD patterns analysis revealed that NPs were formed after calcination, which showed a cubic and hexagonal crystalline structure of (CdO)x (ZnO)1-x NPs. The phase analysis using EDX spectroscopy and FTIR spectroscopy confirmed the presence of Cd and Zn as the original compounds of prepared (CdO)x (ZnO)1-x NP samples. The average particle size of the samples increased from 14 to 33 nm as the concentration of precursor increased from x=0.20 to x=0.80, as observed by TEM results. The surface composition and valance state of the prepared product NPs were determined by X-ray photoelectron spectroscopy (XPS) analyses. Diffuse UV-visible reflectance spectra were used to determine the optical band gap through the Kubelka-Munk equation; the energy band gap was found to decrease for CdO from 2.92 to 2.82 eV and for ZnO from 3.22 to 3.11 eV with increasing x value. Additionally, photoluminescence (PL) spectra revealed that the intensity in PL increased with an increase in particle size. In addition, the antibacterial activity of binary oxide NP was carried out in vitro against Escherichia coli ATCC 25922 Gram (-ve), Salmonella choleraesuis ATCC 10708, and Bacillus subtilis UPMC 1175 Gram (+ve). This study indicated that the zone of inhibition of 21 mm has good antibacterial activity toward the Gram-positive B. subtilis UPMC 1175.
A new 2-arylbenzofuran, sesbagrandiflorain C (1), together with four known compounds, 2-(3,4-dihydroxy-2-methoxyphenyl)-4-hydroxy-6-methoxybenzofuran-3-carbaldehyde (2), 2-(4-hydroxy-2-methoxyphenyl)-5,6-dimethoxybenzofuran-3-carboxaldehyde (3), sesbagrandiflorain A (4) and sesbagrandiflorain B (5), have been isolated from the stem bark of an Indonesian plant, Sesbania grandiflora (L.) Pers. The chemical structure of compound 1 was elucidated by UV, IR, MS, and NMR spectroscopic techniques. The proton and carbon NMR resonances of 1 were also compared with the predicted chemical shifts obtained from DFT quantum mechanical calculations with Gaussian. None of the compounds showed antibacterial activity against Bacillus subtilis, Escherichia coli, Mycobacterium smegmatis, Pseudomonas aeruginosa, and Staphylococcus aureus in an agar diffusion assay. However, sesbagrandiflorains A (4) and B (5) exhibited moderate activity against Mycobacterium tuberculosis H37Rv. In addition, compounds 1 - 5 have moderate cytotoxicity against HeLa, HepG2, and MCF-7 cancer cell lines.
Citrus canker caused by Xanthomonas citri subsp. citri is a disease affecting the yield and fruit quality of lime (Citrus aurantiifolia). This research investigated endophytic bacteria obtained from six healthy Citrus spp. to inhibit the pathogen and to control citrus canker on lime plants. Numbers of the endophytic bacteria isolated from C. aurantifolia, C. hystrix, C. maxima, C. nobilis, C. reticulata and C. sinensis were 28, 25, 29, 42, 12 and 34 isolates, respectively. The selected endophytic bacteria that were effective against X. citri subsp. citri were Bacillus amyloliquefaciens LE109, B. subtilis LE24 and B. tequilensis PO80. The optimum culture medium for an antagonistic effect on the pathogen in B. amyloliquefaciens LE109 and B. tequilensis PO80 was yeast extract peptone dextrose broth, and in B. subtilis LE24 was modified soluble starch broth. To control citrus canker in lime, young expanded leaves of lime plants were aseptically punctured and inoculated with 30 μl of bacterial suspension of the pathogen (108 CFU/ml in 0.85% NaCl) per punctured location. After the pathogenic inoculation for 24 h, the leaves were then inoculated with 30 μl of the selected endophytic bacteria (108 CFU/ml in 0.85% NaCl), and treated with 30 μl of the culture media containing bioactive compounds produced by the selected endophytic bacteria. The leaves inoculated with cell suspensions of B. amyloliquefaciens LE109 or B. subtilis LE24 could completely control citrus canker. However, the leaves inoculated with B. tequilensis PO80 displayed 10% disease incidence. Additionally, the leaves treated with the crude bioactive compounds of B. amyloliquefaciens LE109 or B. subtilis LE24 could completely control citrus canker. Notably, the leaves treated with the crude bioactive compounds of B. tequilensis PO80 displayed 5% disease incidence. The results of this study showed that the Bacillus strains play important roles in the biocontrol of citrus canker in lime.
A series of N'-(substituted benzylidene)-2-(benzo[d]oxazol-3(2H)-yl)acetohydrazide derivatives was synthesized and evaluated for its in vitro antimicrobial and anticancer activities. Antimicrobial activity results revealed that compound 12 was found to be the most potent antimicrobial agent. Results of anticancer study indicated that the synthesized compounds exhibited average anticancer potential. Compound 7 (IC 50 =3.12 µM) and compound 16 (IC 50 =2.88 µM) were found to be most potent against breast cancer (MCF7) cell lines. In conclusion, compound 12 and 16 have the potential to be selected as lead compound for the developing of novel antimicrobial and anticancer agents respectively.
Matched MeSH terms: Bacillus subtilis/drug effects; Bacillus subtilis/growth & development