Bacterium Bacillus Sp Research Articles

Mineral fortification modifies physical and microstructural characteristics of milk gels coagulated by a bacterial enzymatic pool

An enzymatic pool from the Amazonian bacterium Bacillus sp. P7 was used as milk coagulant. Discovery of novel coagulants is of great interest in dairy industry for the development of new textures in cheese. Color, mechanical and microstructural characterization of milk gels induced by the bacterial enzymatic pool was carried out. Effect of mineral fortification on these characteristics was studied. Whiter gels with smaller pore diameters were obtained in the presence of Ca2+ or Mg2+. These characteristics seemed to be influenced by the effect of ionic strength on casein structure which was also evidenced by digital texture features analysis. On the other hand, specific affinity of the assayed cations for milk proteins showed to be important in the development of the mechanical texture of the gels. Firmness and fracture force of milk gels obtained in the presence of Zn2+ or Ca2+ were higher than in the presence of Mg2+ and Na2+.

Optimization, Production, and Partial Purification of Thermostable -Amylase Produced by Marine Bacterium Bacillus sp. NRC12017

α-Amylase biosynthesis was illustrated under stress circumstances of high temperature and high salinity in aerobically cultivated culture of a newly isolated moderately thermophilic bacterium of spore-forming Bacillus sp. NRC12017 in medium containing starch, peptone and yeast extract. Maximum yield (18.41 U/ml) was took place at pH 6.5 with 200 µl inoculum size at 45°C and an incubation time of 3 days. The ideal volume of the fermentation broth was found to be 15 ml in 100 ml Erlenmeyer flask, supplementation of starch at 2.5% and peptone plus yeast extract at 0.7% brought about the greatest production of α-amylase. Two fractions of α-amylase activities, designated FIII and FIV, were refined from culture filtrate utilizing ammonium sulfate (60 and 80%) and showed the main band at 30 KDa. Both fractions had the highest activity at 250 µl starch in a reaction mixture and pH 6.0-7.0. FIII and FIV showed that a temperature of 50 and 55°C and a reaction time of 20 and 30 min were the best available conditions for their activities, respectively, both fractions were stable up to 65°C and the activity was decreased drastically to 3.22 and 3.26 % when heated at 70°C. Concerning pH stability, a broad range of pH stability (5.0-11.0) was obtained by FIII and FIV.

Efektifitas Pestisida Biologis Bacillus Cereus dan Bacillus Megaterium. sebagai Pengendali Spodoptera litura Fabr (Lepidoptera: Noctuidae)

This research was conducted in vitro in the laboratory. Studies conducted in the Laboratory of Bacteriology Department of Plant Pests and Diseases of the Faculty of Agriculture, University of Brawijaya, on the effectiveness of the bacteria Bacillus cereus and Bacillus megaterium as biological pesticides controlling Spodoptera litura. The objective of this study was to determine the effectiveness of the bacteria Bacillus sp as biological control. Effectiveness pesticide was measured by testing the incubation period and mortality in larvae of S. litura instar 3. This study used a completely randomized design (CRD), conducted observations every 6 hours until the larvae dead. The results showed that the percentage of mortality of S. litura reached 94.66% compared to the control, while the incubation time of the bacteria B. cereus cause disease until 29.84 hours. B. cereus and B. megaterium have the ability to incubate third instar larvae of S. litura up to 29.84 hours and caused the death of larvae up to 94.66%. Keywords: Biological pesticides, Bacillus cereus, Bacillus megaterium, entomopatogen, Spodoptera litura

Efektivitas Pemberian Bacillus sp. D2.2 pada Media Teknis Molase terhadap Kualitas Air dan Performa Pertumbuhan Udang Vaname (Litopenaeus vannamei)

Vanamei prawns have fast growth and can reared in high density. it has an impact on water quality deterioration and disruption of survival rates and growth. Various ways to cope has been done, one of them is with probiotic bacteria. The new strain of D2.2 bacteria is thought to be effective of ammonia utilization. Probiotics with local bacteria Bacillus sp. D2.2 is cultured on molasses technical medium to be applied semi-mass. The purpose of this study is to asses the efectivity of Bacillus sp. D2.2 inthe molasses technical medium on water quality and growth performance of vaname prawns (Litopenaeus vannamei). The research was used complete randomized design (RAL) with four treatments, A (Control), B (Application of 5 ppm Bacillus sp. D2.2 cultured in molasses technical medium), C (Application of 10 ppm Bacillus sp. D2.2 cultured in molasses technical medium), D (Application of 15 ppm Bacillus sp. D2.2 cultured in molasses technical medium) were repeated three times each. The results showed no effect on water quality and shrimp survival rate, but absolute growth (W), daily growth rate (GR) and feed conversion ratio (FCR) showed that B and C treatment had better than control.
 Keywords: Vaname shrimp, growth, Bacillus sp. D2.2, molasses technical medium

Symbiotic performance and induction of systemic resistance against Cercospora sojina in soybean plants co-inoculated with Bacillus sp. CHEP5 and Bradyrhizobium japonicum E109.

Soybean is an economically very important crop throughout the word and particularly in Argentina. Soybean yield may be affected by many factors such as the lack of some essential nutrients or pathogens attack. In this work we demonstrated that the co-inoculation of the native biocontrol bacterium Bacillus sp. CHEP5 which induces resistance against Cercospora sojina in soybean and the nitrogen fixing strain Bradyrhizobium japonicum E109, was more effective in reducing frog leaf spot severity than the inoculation of the biocontrol agent alone. Probably, this is related with the increase in the ability to form biofilm when both bacteria are growing together. Furthermore, Bacillus sp. CHEP5 inoculation did not affect Bradyrhizobium japonicum E109 symbiotic behavior and flavonoids composition of root exudates in pathogen challenged plants. These results suggest that co-inoculation of plants with rhizobia and biocontrol agents could be a strategy to improve soybean production in a sustainable system.

Crystal Structure and Functional Characterization of a Cytochrome P450 (BaCYP106A2) from Bacillus sp. PAMC 23377.

Bacterial cytochrome P450 (CYP) steroid hydroxylases are effectively useful in the pharmaceutical industry for introducing hydroxyl groups to a wide range of steroids. We found a putative CYP steroid hydroxylase (BaCYP106A2) from the bacterium Bacillus sp. PAMC 23377 isolated from Kara Sea of the Arctic Ocean, showing 94% sequence similarity with BmCYP106A2 (Bacillus megaterium ATCC 13368). In this study, soluble BaCYP106A2 was overexpressed to evaluate its substrate-binding activity. The substrate affinity (Kd value) to 4-androstenedione was 387 ± 37 µM. Moreover, the crystal structure of BaCYP106A2 was determined at 2.7 Å resolution. Structural analysis suggested that the α8-α9 loop region of BaCYP106A2 is intrinsically mobile and might be important for initial ligand binding. The hydroxyl activity of BaCYP106A2 was identified using in vitro enzyme assays. Its activity was confirmed with two kinds of steroid substrates, 4-androstenedione and nandrolone, using chromatography and mass spectrometry methods. The main products were monohydroxylated compounds with high conversion yields. This is the second study on the structure of CYP106A steroid hydroxylases, and should contribute new insight into the interactions of bacterial CYP106A with steroid substrates, providing baseline data for studying the CYP106A steroid hydroxylase from the structural and enzymatic perspectives.

Pengaruh Pemaparan Medan Magnet Pada Media Mandels Yang Dimodifikasi Terhadap Pertumbuhan Dan Aktivitas Enzim Selulase Bacillus Sp.

Enzymes can undergo structural changes due to various environmental factor such as pH,temperature, electricity and magnetic fields. The purpose of this study was to investigate theeffect of magnetic field strength and duration of exposure to the modified of media Mandelsused as a culture medium of Bacillus sp. on the growth and activity of cellulase enzyme fromthe bacterium Bacillus sp. Factorial study group were prepared using the method of fullrandomized design (RKLT). The first factor is the strong magnetic field consists of 4 levels ofmagnetic field strength treatment: 0 mT; 0,1 mT; 0,2 mT; 0.3 mT. The second faktor is theprolonged exposure consisting of 4 levels prolonged exposure treatments 0, 10, 20, 30minutes. each unit treatment was repeated 3 times. The results showed that exposure tomagnetic fields on modified of media Mandels causes a decrease in the relative activity ofthe enzyme cellulase and relative growth of Bacillus sp.Keywords: cellulase enzyme, magnetic field, Mandels modification, Bacillus sp.

UJI EFEKTIVITAS EKSTRAK KUNYIT SEBAGAI ANTIBAKTERI DALAM PERTUMBUHAN Bacillus sp dan Shigella dysentriae SECARA IN VITRO

Turmeric is a spice plants that acts as an antibacterial, because it contains a variety of compounds including curcumin and essential oil. Essential oils can be used as an antibacterial because it contains hydroxyl and carbonyl functional group which is phenols derivative. Te phenol derivatives will interact with the bacterial cell wall, then absorbed and penetrated into the bacterial cell, causing precipitation and denaturation of proteins, the result will lyse the bacterial cell membrane, while the antibacterial activity of curcuminis by inhibiting bacterial cell proliferation. Turmeric has launched a pharmacological effect, lowering the fat content, asthma , hepatitis , anti- gall , anti- inflammatory , anti- diarrhea , and act as anti-inflammatory or anti-inflammatory. Turmeric has antibacterial properties of curcumin and essential oil that is capable of inhibiting the growth of bacteria that causes diarrhea and Shigelladysenteriae Bacillus sp. Tis study was conducted to determine the effectiveness of turmeric extract on the growth of Bacillus sp and Shigella dysenteriae, with various concentrations of 15%, 30%, 50%, 75%, and 100% by the well diffusion method. Based on theinhibition zone measurement of bacteria Bacillus sp and Shigella dysenteriaethe results were weak category, for the bacteria Bacillus sp with a concentration of 15 % , 30 % , 50 % , 75 % , and 100 % with a diameter of 11 ; 12.3 ; 13 , 3 ; 13.7 ; 14.7 mm, while for the bacteria Shigella dysenteriae with the same concentration has a diameter of 10.3 ; 11.7 ; 12.3 ; 13.3 , and 14.7 mm. Te conclusion of the study is that the antibacterial activity of turmeric extract is more effective against the bacteria Bacillus spthan against bacteria Shigella dysenteriae, although the difference was not signifcant.

Anti-Pigmentary Effect of (-)-4-Hydroxysattabacin from the Marine-Derived Bacterium Bacillus sp.

Bioactivity-guided isolation of a crude extract from a culture broth of Bacillus sp. has led to the isolation of (-)-4-hydroxysattabacin (1). The inhibitory effect of (-)-4-hydroxysattabacin (1) was investigated on melanogenesis in the murine melanoma cell line, B16F10, and human melanoma cell line, MNT-1, as well as a pigmented 3D-human skin model. (-)-4-Hydroxysattabacin treatment decreased melanin contents in a dose-dependent manner in α-melanocyte stimulating hormone (α-MSH)-stimulated B16F10 cells. Quantitative real time PCR (qRT–PCR) demonstrated that treatment with (-)-4-hydroxysattabacin down-regulated several melanogenic genes, including tyrosinase, tyrosinase-related protein 1 (TRP-1), and tyrosinase-related protein 2 (TRP-2) while their enzymatic activities were unaffected. The anti-melanogenic effects of (-)-4-hydroxysattabacin were further demonstrated in a pigmented 3D human epidermal skin model, MelanodermTM, and manifested as whitening and regression of melanocyte activation in the tissue.

Impacts of Hydrogen Peroxide and Copper Sulfate on the Control of Microcystis aeruginosa and MC-LR and the Inhibition of MC-LR Degrading Bacterium Bacillus sp.

Laboratory batch experiments were carried out to evaluate the impacts of H2O2 and copper sulfate on M. aeruginosa PCC7820, microcystin-LR (MC-LR) and its degrading bacteria Bacillus sp., previously isolated from Hulupi Lake in Taiwan. The study shows that 3 mg·L−1 hydrogen peroxide removed only 9% M. aeruginosa within seven days of exposure, from an initial cell concentration of 2 × 106 cells/mL. With copper sulfate, a concentration of 2 mg·L−1 removed 99% M. aeruginosa cells, but showed negligible efficacy in removing 0.05 mg·L−1 MC-LR. At a higher dosage, 20 mg·L−1 H2O2 led to 40% and 95% removal, respectively for MC-LR and M. aeruginosa cells. Copper sulfate and H2O2 were both lethal to Bacillus sp. population, with mortality rate constants of k = 0.04 h−1 and 0.03 h−1 under 1 mg·L−1 copper sulfate and 5 mg·L−1 H2O2, respectively. H2O2 is competitive in terms of cost, with a capability of degrading organic compounds with the assistance of ultraviolet (UV) light, and it may be considered as an alternative algaecide to copper sulfate in reservoirs for algae growth control.

Biodegradation and kinetic study of benzene in bioreactor packed with PUF and alginate beads and immobilized with Bacillus sp. M3

Benzene removal in free and immobilized cells on polyurethane foam (PUF) and polyvinyl alcohol (PVA)-alginate beads was studied using an indigenous soil bacterium Bacillus sp. M3 isolated from petroleum-contaminated soil. The important process parameters (pH, temperature and inoculums size) were optimized and found to be 7, 37°C and 6.0×108CFU/mL, respectively. Benzene removals were observed to be 70, 84 and 90% within 9days in a free cell, immobilized PVA-alginate beads and PUF, respectively under optimum operating conditions. FT-IR and GC–MS analysis confirm the presence of phenol, 1,2-benzenediol, hydroquinone and benzoate as metabolites. The important kinetic parameter ratios (µmax/Ks; L/mg·day) calculated using Monod model was found to be 0.00123 for free cell, 0.00159 for immobilized alginate beads and 0.002016 for immobilized PUF. Similarly inhibition constants (Ki; mg/L) calculated using Andrew-Haldane model was found to be 435.84 for free cell, 664.25 for immobilized alginate beads and 724.93 for immobilized PUF.

Recycling of carbon dioxide by free air CO2 enriched (FACE) Bacillus sp. SS105 for enhanced production and optimization of biosurfactant

Carbon dioxide utilizing bacterium Bacillus sp. SS105 was isolated from FACE (free air CO2 enriched) sample. The strain was grown in shake flask containing minimal salt medium with 50mM NaHCO3 as autotrophic carbon source and molasses as a low cost byproduct for mixotrophic growth. Carbon dioxide sequestration property of Bacillus sp. SS105 was determined by enzyme assay of carbonic anhydrase and ribulose-1, 5-bisphosphate carboxylase/oxygenase (RuBisCO). Along with CO2 sequestration this strain produced biosurfactant and its characterization by FTIR and 1H NMR indicated lipopeptide nature. Optimization of process parameter along with nutrient sources for higher biosurfactant production was done by Response Surface Methodology (RSM). Under optimized conditions, the yield of biosurfactant and biomass was 2.65 and 2.78gL−1 respectively. The study revealed simultaneous CO2 sequestration and biosurfactant production by Bacillus sp. SS105.

Lactose-inducted production of a complete lignocellulolytic enzyme system by a novel bacterium Bacillus sp. BS-5 and its application for saccharification of alkali-pretreated corn cob

In this study, with combined carboxymethyl cellulose agar plate, xylan agar plate and filter paper hydrolysis assay, a novel cellulase and xylanase-producing strain identified as Bacillus sp. was isolated. Using lactose as the only carbon source, a complete and balanced lignocellulolytic enzyme system containing at least endoglucanase (9.6 U/ml), exoglucanase (0.8 U/ml), Fpase (1.4 U/ml), xylanase (3.8 U/ml) and β-glucosidase (1.2 U/ml) was produced. Interestingly, a zymogram of the crude culture supernatant displayed a multifunctional lignocellulolytic enzyme system including at least four bonds with both endoglucanase activity and xylanase activity at 21.2, 23.8, 28.9 and 31.2 kDa, respectively, indicating that these enzymes might be bifunctional. More gratifyingly, according to the binding affinity analysis and scanning electron microscopy, the crude enzyme complex produced by strain BS-5 was capable of hydrolyzing not only pure insoluble polysaccharides, but also agricultural residues such as corn cob. At 5% substrate concentration and 20 FPU/g enzyme loading, the reducing sugar was 350.8 mg/g of alkali-pretreated corn cob after 72 h enzymatic hydrolysis. These results suggested that this strain could be a good candidate for the development of a more cost-effective and efficient lignocellulolytic enzyme cocktail for the saccharification of lignocellulosic biomass.

Probiotic efficiency of Bacillus sp. in Labeo rohita challenged by Aeromonas hydrophila : assessment of stress profile, haemato-biochemical parameters and immune responses

Fish are always susceptible to a wide variety of deadly pathogens which cause a huge loss in aquaculture industries. In this investigation, we have demonstrated the in vivo probiotic efficiency of Bacillus sp. MVF1 (GenBank Acc. No. KP256503) in Labeo rohita challenged with pathogenic strain of Aeromonas hydrophila (MTCC 1739). To check the probiotic potential of the selected bacterial strain, fish were divided into four groups: control, D1, D2 and D3. A total of 100 days (70 days probiotic feeding + 71th day sampling and 28 days challenged test + 29th day sampling) of feeding trial was conducted. To establish the probiotic potential of Bacillus sp. MVF1, certain haematological parameters (haemoglobin, total erythrocyte and leucocyte count), serum biochemical parameters (total protein, albumin and globulin), immune parameters (serum lysozyme and total IgM levels) and hepatic stress profile (malondialdehyde production, superoxide dismutase and catalase activity) have been measured. Our results demonstrated that red blood cell number, white blood cell number and haemoglobin content were much higher in D2 group fish compared to other groups and control fish. Similarly, total protein contents, albumin concentration, globulin concentration, lysozyme activity and IgM production were also recorded to be highest in D2 group fish. This finding clearly indicated the probiotic potential of Bacillus sp. MVF1 in L. rohita. Furthermore, our results also demonstrated that 1 × 107 CFU g−1 feed (D2) provides better immunity compared to 1 × 105 (D1) and 1 × 109 (D3). Due to beneficial effects, the bacterium Bacillus sp. MVF1 might be useful in aquaculture industries to reduce the disease susceptibility.

Isolation and characterization of mannanase, xylanase, and cellulase from marine bacteria Bacillus sp.

Yopi, Djohan AC, Rahmani N, Jannah AM. 2017. Isolation and characterization of mannanase, xylanase, and cellulase from marine bacteria Bacillus sp. Biofarmasi (Rumphius J Nat Prod Biochem) 15: 15-20. Isolation, identification, and characterization of mannanase, xylanase and cellulase producing indigenous marine bacteria have been conducted from total 20 isolates. Based on 16S rDNA sequence analysis, three potential isolates are obtained and identified as Bacillus subtilis (M8), Bacillus tequilensis (X4) and Bacillus cereus (C9). The potential strains M8, X4 and C9 can produce mannanase, xylanase and cellulase activities such as 9.5 U/mL; 0.36U/mL;0.56U/mL with optimum pH and temperature 6.0;50oC, 5.5;70oC and 8;50oC, respectively. Based on the TLC analysis, mannanase from M8 and xylanase from X4 has potential to hydrolyzed mannan and xylan for producing oligosaccharides with size around tri-hexasaccharides as a main product.

Mitigation Effect of Drought Stress by Plant Growth-promoting Bacterium Bacillus sp. SB19 on Kale Seedlings in Greenhouse

Mitigation Effect of Drought Stress by Plant Growth-promoting Bacterium Bacillus sp. SB19 on Kale Seedlings in Greenhouse

The optimal of xylanase production conditions using empty fruit bunch raw biomass by marine bacterium Bacillus sp LBF-001

Several xylanases have been studied recently, but few xylanases are from marine microorganisms have been reported. Marine bacterium Bacillus sp. LBF-001 was isolated from Pari Island Kepulauan Seribu in Indonesia. The purposes of this study are to identify of 16S rDNA gene from marine bacterium LBF-001 and to optimize medium conditions including kind and concentration of biomass, nitrogen source, pH and temperature. With 16S rDNA gene analysis that LBF-001 isolate have 99% identity with Bacillus pumilus HT-Z4-B2 (KJ526885). Fermentation for producing xylanases was done by using several agricultural residues under solid-state fermentation (SSF). The optimum condition for xylanase production by isolate Bacillus sp. LBF-001 are 2.5% empty fruit bunch, 0.6% lactose broth, pH values was 6.5, temperature condition was 30oC, under submerged fermentation with shaking at 150 rpm for 48 hours fermentation and giving the xylanase activity 10.85 U/mL.

Biosynthesis and Characterization of Poly3-hydroxyalkanote (PHA) from Newly Isolated Bacterium Bacillus sp. AZR-1

Bacterial poly3-hydroxyalkanotes are the polyesters that are accumulated in conditions of un-balanced nutrients. Soil samples were collected from Hilly areas of Muzaffarabad, and oil processing area of Lahore, Pakistan to isolate PHA producing bacteria. The bacteria were screened on Sudan black B and Nile blue A staining methodologies. An efficient PHA producing strain AZR-1 was isolated and characterized morphologically and biochemically. The strain was identified as Bacillus sp. AZR-1 on the genetic basis of 16S rRNA gene sequence. Bacillus sp. AZR-1 produced 40% PHA contents when grown on N-limited mineral salt medium supplemented with glucose in shake flasks. Starch and sodium gluconate were found to be other putative carbon sources for PHA production as the bacterium produced 22 and 17% PHA on these sources. FTIR confirmed the presence of scl-PHA monomers in the polymer extracted from the bacterium. The PHA synthase gene (phaC) was partially amplified and sequenced which showed maximum similarity with the PHA synthase of Bacillus weihenstephanensis KBAB4.

Parameter’s optimization and kinetics study of α-amylase enzyme of Bacillus sp. MB6 isolated from vegetable waste

Abstract α-Amylase, a very critical enzyme for hydrolysis of starch into simple sugar and it has various applications in industrial settings. This study reports the identification of Bacillus sp. MB6 which produces increased amount of enzyme from less required resources. To optimize the yield of enzyme, we used various combinations of parameters. The most optimized conditions for production of amylase enzyme from the bacterium Bacillus sp. MB6 are pH of 6, temperature of 37 °C, and incubation period of 48 h. Condition of enzymatic activity were also examined and the results show that pH of 6, a temperature of 55 °C, and a reaction time of 30 min are the best available conditions for its activity. Purification of enzyme by 1.63 fold enhanced the specific activity of enzyme based upon its activity analysis as compared with unpurified enzyme. Enzyme kinetics studies show the Michaelis constant (Km) to be 5.45 mg/ml and maximum velocity of the reaction (Vmax) to be 24.15 mg/ml/min. In conclusion, we report enzyme production and purification methodology that exhibit better yield of alpha-amylase for commercial applications.

Chitinolytic Bacillus-Mediated Induction of Jasmonic Acid and Defense-Related Proteins in Soybean (Glycine max L. Merrill) Plant Against Rhizoctonia solani and Fusarium oxysporum

In the present research, bacterial-mediated elicitation of induced systemic resistance in the soybean plant was studied. The main objective was in vitro analysis of jasmonic acid and different defense-related enzymes in soybean plants primed with bacterium Bacillus sp. SJ-5 against the fungal pathogen Rhizoctonia solani and Fusarium oxysporum. In the different assays conducted, Bacillus sp. SJ-5 showed strong antifungal activity against R. solani and F. oxysporum showing 45 and 63 % growth inhibition, respectively. Strain SJ-5 was found to be positive for the cell wall-degrading enzymes chitinase, protease, and β-1,3-glucanase, and cell-free supernatant was found with significant fungal growth inhibitory activity. Different defense-related enzymes, namely lipoxygenase, phenylalanine ammonia-lyase, peroxidase, polyphenol oxidase, and β-1,3-glucanase in the different parts of Glycine max L. Merrill were reported to be highest on the 8th day after challenge inoculation and expressed significantly in the root tissue. GC–MS analysis of jasmonic acid (JA) revealed the highest JA accumulation in bacterized soybean plant root tissue challenged with R. solani and F. oxysporum, which was 91.2 and 99.84 %, respectively, with respect to control. In the SJ-5-primed root tissue, phenolic content was highest upon challenge inoculation of R. solani and F. oxysporum with 30.47 ± 0.97 and 32.4 ± 0.3 mg/g fresh weight, respectively. In summary, the present investigation revealed a role for the bacterial isolate Bacillus sp. SJ-5 in soybean plant growth promotion and enhanced protection against R. solani and F. oxysporum by elicitation of defense-related enzymes.