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Inoculation of Bacillus sphaericus UPMB-10 to young oil palm and measurement of its uptake of fixed nitrogen using the ¹⁵N isotope dilution technique

Zakry FA, Shamsuddin ZH, Abdul Rahim K, Zawawi Zakaria Z, Abdul Rahim A

Microbes Environ, 2012;27(3):257-62.
PMID: 22446306

There are increasing applications of diazotrophic rhizobacteria in the sustainable agriculture system. A field experiment on young immature oil palm was conducted to quantify the uptake of N derived from N₂ fixation by the diazotroph Bacillus sphaericus strain UPMB-10, using the ¹⁵N isotope dilution method. Eight months after ¹⁵N application, young immature oil palms that received 67% of standard N fertilizer application together with B. sphaericus inoculation had significantly lower ¹⁵N enrichment than uninoculated palms that received similar N fertilizers. The dilution of labeled N served as a marker for the occurrence of biological N₂ fixation. The proportion of N uptake that was derived from the atmosphere was estimated as 63% on the whole plant basis. The inoculation process increased the N and dry matter yields of the palm leaflets and rachis significantly. Field planting of young, immature oil palm in soil inoculated with B. sphaericus UPMB-10 might mitigate inorganic fertilizer-N application through supplementation by biological nitrogen fixation. This could be a new and important source of nitrogen biofertilizer in the early phase of oil palm cultivation in the field.

Matched MeSH terms: Bacillus/metabolism*
Fulltext Binary Toxin Subunits of Lysinibacillus sphaericus Are Monomeric and Form Heterodimers after In Vitro Activation

Surya W, Chooduang S, Choong YK, Torres J, Boonserm P

PLoS One, 2016;11(6):e0158356.
PMID: 27341696 DOI: 10.1371/journal.pone.0158356

The binary toxin from Lysinibacillus sphaericus has been successfully used for controlling mosquito-transmitted diseases. An activation step shortens both subunits BinA and BinB before their interaction with membranes and internalization in midgut cells, but the precise role of this activation step is unknown. Herein, we show conclusively using three orthogonal biophysical techniques that protoxin subunits form only monomers in aqueous solution. However, in vitro activated toxins readily form heterodimers. This oligomeric state did not change after incubation of these heterodimers with detergent. These results are consistent with the evidence that maximal toxicity in mosquito larvae is achieved when the two subunits, BinA and BinB, are in a 1:1 molar ratio, and directly link proteolytic activation to heterodimerization. Formation of a heterodimer must thus be necessary for subsequent steps, e.g., interaction with membranes, or with a suitable receptor in susceptible mosquito species. Lastly, despite existing similarities between BinB C-terminal domain with domains 3 and 4 of pore-forming aerolysin, no aerolysin-like SDS-resistant heptameric oligomers were observed when the activated Bin subunits were incubated in the presence of detergents or lipidic membranes.

Matched MeSH terms: Bacillus/metabolism*
Fulltext Biodegradation of palm kernel cake by cellulolytic and hemicellulolytic bacterial cultures through solid state fermentation

Alshelmani MI, Loh TC, Foo HL, Lau WH, Sazili AQ

ScientificWorldJournal, 2014;2014:729852.
PMID: 25019097 DOI: 10.1155/2014/729852

Four cellulolytic and hemicellulolytic bacterial cultures were purchased from the Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Culture (DSMZ) and the American Type Culture Collection (ATCC). Two experiments were conducted; the objective of the first experiment was to determine the optimum time period required for solid state fermentation (SSF) of palm kernel cake (PKC), whereas the objective of the second experiment was to investigate the effect of combinations of these cellulolytic and hemicellulolytic bacteria on the nutritive quality of the PKC. In the first experiment, the SSF was lasted for 12 days with inoculum size of 10% (v/w) on different PKC to moisture ratios. In the second experiment, fifteen combinations were created among the four microbes with one untreated PKC as a control. The SSF lasted for 9 days, and the samples were autoclaved, dried, and analyzed for proximate analysis. Results showed that bacterial cultures produced high enzymes activities at the 4th day of SSF, whereas their abilities to produce enzymes tended to be decreased to reach zero at the 8th day of SSF. Findings in the second experiment showed that hemicellulose and cellulose was significantly (P < 0.05) decreased, whereas the amount of reducing sugars were significantly (P < 0.05) increased in the fermented PKC (FPKC) compared with untreated PKC.

Matched MeSH terms: Bacillus/metabolism*; Paenibacillus/metabolism*
Isotherm equilibria of Mn²⁺ biosorption in drinking water treatment by locally isolated Bacillus species and sewage activated sludge

Hasan HA, Abdullah SR, Kofli NT, Kamarudin SK

J Environ Manage, 2012 Nov 30;111:34-43.
PMID: 22813857 DOI: 10.1016/j.jenvman.2012.06.027

Manganese (Mn(2+)) is one of the inorganic contaminant that causes problem to water treatment and water distribution due to the accumulation on water piping systems. In this study, Bacillus sp. and sewage activated sludge (SAS) were investigated as biosorbents in laboratory-scale experiments. The study showed that Bacillus sp. was a more effective biosorbent than SAS. The experimental data were fitted to the Langmuir (Langmuir-1 & Langmuir-2), Freundlich, Temkin, Dubinin-Radushkevich (D-R) and Redlich-Peterson (R-P) isotherms to obtain the characteristic parameters of each model. Mn(2+) biosorption by Bacillus sp. was found to be significantly better fitted to the Langmuir-1 isotherm than the other isotherms, while the D-R isotherm was the best fit for SAS; i.e., the χ(2) value was smaller than that for the Freundlich, Temkin, and R-P isotherms. According to the evaluation using the Langmuir-1 isotherm, the maximum biosorption capacities of Mn(2+) onto Bacillus sp. and SAS were 43.5 mg Mn(2+)/g biomass and 12.7 mg Mn(2+)/g biomass, respectively. The data fitted using the D-R isotherm showed that the Mn(2+) biosorption processes by both Bacillus sp. and SAS occurred via the chemical ion-exchange mechanism between the functional groups and Mn(2+) ion.

Matched MeSH terms: Bacillus/metabolism*
Fulltext Application of potential phosphate-solubilizing bacteria and organic acids on phosphate solubilization from phosphate rock in aerobic rice

Panhwar QA, Jusop S, Naher UA, Othman R, Razi MI

ScientificWorldJournal, 2013;2013:272409.
PMID: 24288473 DOI: 10.1155/2013/272409

A study was conducted at Universiti Putra Malaysia to determine the effect of phosphate-solubilizing bacteria (PSB) and organic acids (oxalic & malic) on phosphate (P) solubilization from phosphate rock (PR) and growth of aerobic rice. Four rates of each organic acid (0, 10, 20, and 30 mM), and PSB strain (Bacillus sp.) were applied to aerobic rice. Total bacterial populations, amount of P solubilization, P uptake, soil pH, and root morphology were determined. The results of the study showed significantly high P solubilization in PSB with organic acid treatments. Among the two organic acids, oxalic acid was found more effective compared to malic acid. Application of oxalic acid at 20 mM along with PSB16 significantly increased soluble soil P (28.39 mg kg(-1)), plant P uptake (0.78 P pot(-1)), and plant biomass (33.26 mg). Addition of organic acids with PSB and PR had no influence on soil pH during the planting period. A higher bacterial population was found in rhizosphere (8.78 log10 cfu g(-1)) compared to the nonrhizosphere and endosphere regions. The application of organic acids along with PSB enhanced soluble P in the soil solution, improved root growth, and increased plant biomass of aerobic rice seedlings without affecting soil pH.

Matched MeSH terms: Bacillus/metabolism*
Biological treatment of sewage treatment plant sludge by pure bacterial culture with optimum process conditions in a stirred tank bioreactor

Alam MZ, Muyibi SA, Jamal P

J Environ Sci Health A Tox Hazard Subst Environ Eng, 2007 Sep;42(11):1671-9.
PMID: 17849310

Biological treatment of sewage treatment plant (STP) sludge by potential pure bacterial culture (Bacillus sp.) with optimum process conditions for effective biodegradation and bioseparation was carried out in the laboratory. The effective and efficient bioconversion was evaluated with the treatment of pure bacterial culture and existing microbes (uninnoculated) in sludge. The optimum process conditions i.e., temperature, 40 degrees C; pH, 6; inoculum, 5% (v/v); aeration, 1 vvm; agitation speed, 50 rpm obtained from the previous studies with chemical oxygen demand COD at 30 mgL(-1) were applied for the biological treatment of sludge. The results indicated that pure bacterial culture (Bacillus sp.) showed higher degradation and separation of treated sludge compared to treatment with the existing mixed microbes in a stirred tank bioreactor. The treated STP sludge by potential pure bacterial culture and existing microbes gave 30% and 11%; 91.2% and 59.1; 88.5% and 52.3%; 98.4% and 51.3%; 96.1% and 75.2%; 99.4% and 72.8% reduction of total suspended solids (TSS, biosolids), COD, soluble protein, turbidity, total dissolved solids (TDS) and specific resistance to filtration (SRF), respectively within 7 days of treatment. The pH was observed at 6.5 and 4 during the treatment of sludge by pure culture and existing microbes, respectively.

Matched MeSH terms: Bacillus/metabolism*
Biodegradation of hydrocarbon contamination by immobilized bacterial cells

Rahman RN, Ghaza FM, Salleh AB, Basri M

J Microbiol, 2006 Jun;44(3):354-9.
PMID: 16820766

This study examined the capacity of immobilized bacteria to degrade petroleum hydrocarbons. A mixture of hydrocarbon-degrading bacterial strains was immobilized in alginate and incubated in crude oil-contaminated artificial seawater (ASW). Analysis of hydrocarbon residues following a 30-day incubation period demonstrated that the biodegradation capacity of the microorganisms was not compromised by the immobilization. Removal of n-alkanes was similar in immobilized cells and control cells. To test reusability, the immobilized bacteria were incubated for sequential increments of 30 days. No decline in biodegradation capacity of the immobilized consortium of bacterial cells was noted over its repeated use. We conclude that immobilized hydrocarbon-degrading bacteria represent a promising application in the bioremediation of hydrocarbon-contaminated areas.

Matched MeSH terms: Bacillus/metabolism
Fulltext Biosurfactant Production by Bacillus salmalaya for Lubricating Oil Solubilization and Biodegradation

Dadrasnia A, Ismail S

Int J Environ Res Public Health, 2015 Aug;12(8):9848-63.
PMID: 26295402 DOI: 10.3390/ijerph120809848

This study investigated the capability of a biosurfactant produced by a novel strain of Bacillus salmalaya to enhance the biodegradation rates and bioavailability of organic contaminants. The biosurfactant produced by cultured strain 139SI showed high physicochemical properties and surface activity in the selected medium. The biosurfactant exhibited a high emulsification index and a positive result in the drop collapse test, with the results demonstrating the wetting activity of the biosurfactant and its potential to produce surface-active molecules. Strain 139SI can significantly reduce the surface tension (ST) from 70.5 to 27 mN/m, with a critical micelle concentration of 0.4%. Moreover, lubricating oil at 2% (v/v) was degraded on Day 20 (71.5). Furthermore, the biosurfactant demonstrated high stability at different ranges of salinity, pH, and temperature. Overall, the results indicated the potential use of B. salmalaya 139SI in environmental remediation processes.

Matched MeSH terms: Bacillus/metabolism*
Encapsulation of Bacillus salmalaya 139SI using double coating biopolymer technique

Vejan P, Abdullah R, Khadiran T, Ismail S

Lett Appl Microbiol, 2019 Jan;68(1):56-63.
PMID: 30339728 DOI: 10.1111/lam.13088

Sustainable crop production for a rapidly growing human population is one of the current challenges faced by the agricultural sector. However, many of the chemical agents used in agriculture can be hazardous to humans, non-targeted organism and environment. Plant growth promoting rhizobacteria have demonstrated a role in promoting plant growth and health under various stress conditions including disease. Unfortunately, bacterial viability degrades due to temperature and other environmental factors (Bashan et al., Plant Soil 378: 1-33, 2014). Encapsulation of bacteria into core-shell biopolymers is one of the promising techniques to overcome the problem. This study deals with the encapsulation of Bacillus salmalaya 139SI using simple double coating biopolymer technique which consist of brown rice protein/alginate and 0·5% low molecular weight chitosan of pH 4 and 6. The influence of biopolymer to bacteria mass ratio and the chitosan pH on the encapsulation process, physic-chemical, morphology and bioactivity properties of encapsulated B. salmalaya 139SI have been studied systematically. Based on the analysis of physico-chemical, morphology and bioactivity properties, B. salmalaya 139S1 encapsulated using double coating encapsulation technology has promising viability pre- and postfreeze-drying with excellent encapsulation yields of 99·7 and 89·3% respectively. SIGNIFICANCE AND IMPACT OF THE STUDY: The need of a simple yet effective way of encapsulating plant growth promoting rhizobacteria is crucial to further improve their benefits to global sustainable agriculture practice. Effective encapsulation allows for protection, controlled release and function of the micro-organism, as well as providing a longer shelf life for the product. This research report offers an innovative yet simple way of encapsulating using double coating technology with environmentally friendly biopolymers that could degrade and provide nutrients when in soil. Importantly, the bioactivity of the bacteria is maintained upon encapsulation.

Matched MeSH terms: Bacillus/metabolism*
Fulltext Optimal reduction of chemical oxygen demand and NH3-N from landfill leachate using a strongly resistant novel Bacillus salmalaya strain

Dadrasnia A, Azirun MS, Ismail SB

BMC Biotechnol, 2017 Nov 28;17(1):85.
PMID: 29179747 DOI: 10.1186/s12896-017-0395-9

BACKGROUND: When the unavoidable waste generation is considered as damaging to our environment, it becomes crucial to develop a sustainable technology to remediate the pollutant source towards an environmental protection and safety. The development of a bioengineering technology for highly efficient pollutant removal is this regard. Given the high ammonia nitrogen content and chemical oxygen demand of landfill leachate, Bacillus salmalaya strain 139SI, a novel resident strain microbe that can survive in high ammonia nitrogen concentrations, was investigated for the bioremoval of ammonia nitrogen from landfill leachate. The treatability of landfill leachate was evaluated under different treatment parameters, such as temperature, inoculum dosage, and pH.
RESULTS: Results demonstrated that bioaugmentation with the novel strain can potentially improve the biodegradability of landfill leachate. B. salmalaya strain 139SI showed high potential to enhance biological treatment given its maximum NH3-N and COD removal efficiencies. The response surface plot pattern indicated that within 11 days and under optimum conditions (10% v/v inoculant, pH 6, and 35 °C), B. salmalaya strain139SI removed 78% of ammonia nitrogen. At the end of the study, biological and chemical oxygen demands remarkably decreased by 88% and 91.4%, respectively. Scanning electron microscopy images revealed that ammonia ions covered the cell surface of B. salmalaya strain139SI.
CONCLUSIONS: Therefore, novel resistant Bacillus salmalaya strain139SI significantly reduces the chemical oxygen demand and NH3-N content of landfill leachate. Leachate treatment by B. salmalaya strain 139SI within 11 days.

Matched MeSH terms: Bacillus/metabolism*
Fulltext Production and Characterization of a Bioflocculant Produced by Bacillussalmalaya 139SI-7 and Its Applications in Wastewater Treatment

Abu Tawila ZM, Ismail S, Dadrasnia A, Usman MM

Molecules, 2018 Oct 18;23(10).
PMID: 30340415 DOI: 10.3390/molecules23102689

The production, optimization, and characterization of the bioflocculant QZ-7 synthesized by a novel Bacillus salmalaya strain 139SI isolated from a private farm soil in Selangor, Malaysia, are reported. The flocculating activity of bioflocculant QZ-7 present in the selected strain was found to be 83.3%. The optimal culture for flocculant production was achieved after cultivation at 35.5 °C for 72 h at pH 7 ± 0.2, with an inoculum size of 5% (v/v) and sucrose and yeast extract as carbon and nitrogen sources. The maximum flocculating activity was found to be 92.6%. Chemical analysis revealed that the pure bioflocculant consisted of 79.08% carbohydrates and 15.4% proteins. The average molecular weight of the bioflocculant was calculated to be 5.13 × 10⁵ Da. Infrared spectrometric analysis showed the presence of carboxyl (COO-), hydroxyl (-OH), and amino (-NH₂) groups, polysaccharides and proteins. The bioflocculant QZ-7 exhibited a wide pH stability range from 4 to 7, with a flocculation activity of 85% at pH 7 ± 0.2. In addition, QZ-7 was thermally stable and retained more than 80% of its flocculating activity after being heated at 80 °C for 30 min. SEM analysis revealed that QZ-7 exhibited a clear crystalline brick-shaped structure. After treating wastewater, the bioflocculant QZ-7 showed significant flocculation performance with a COD removal efficiency of 93%, whereas a BOD removal efficiency of 92.4% was observed in the B. salmalaya strain 139SI. These values indicate the promising applications of the bioflocculant QZ-7 in wastewater treatment.

Matched MeSH terms: Bacillus/metabolism
Entrapment of porous cross-linked enzyme aggregates of maltogenic amylase from Bacillus lehensis G1 into calcium alginate for maltooligosaccharides synthesis

Nawawi NN, Hashim Z, Rahman RA, Murad AMA, Bakar FDA, Illias RM

Int J Biol Macromol, 2020 May 01;150:80-89.
PMID: 32035147 DOI: 10.1016/j.ijbiomac.2020.02.032

Maltooligosaccharides (MOSs) are emerging oligosaccharides in food-based applications and can be synthesized through the enzymatic synthesis of maltogenic amylase from Bacillus lehensis G1 (Mag1). However, the lack of enzyme stability makes this approach unrealistic for industrial applications. The formation of cross-linked enzyme aggregates (CLEAs) is a promising tool for improving enzyme stability, and the substrate accessibility problem of CLEA formation was overcome by the addition of porous agents to generate porous CLEAs (p-CLEAs). However, p-CLEAs exhibited high enzyme leaching and low solvent tolerance. To address these problems, p-CLEAs of Mag1 (Mag1-p-CLEAs) were entrapped in calcium alginate beads (CA). Mag1-p-CLEAs-CA prepared with 2.5% (w/v) sodium alginate and 0.6% (w/v) calcium chloride yielded 53.16% (17.0 U/mg) activity and showed a lower deactivation rate and longer half-life than those of entrapped free Mag1 (Mag1-CA) and entrapped non-porous Mag1-CLEAs (Mag1-CLEAs-CA). Moreover, Mag1-p-CLEAs-CA exhibited low enzyme leaching and high tolerance in various solvents compared to Mag1-p-CLEAs. A kinetic study revealed that Mag1-p-CLEAs-CA exhibited relatively high affinity towards beta-cyclodextrin (β-CD) (Km = 0.62 mM). MOSs (300 mg/g) were synthesized by Mag1-p-CLEAs-CA at 50 °C. Finally, the reusability of Mag1-p-CLEAs-CA makes them as a potential biocatalyst for the continuous synthesis of MOSs.

Matched MeSH terms: Bacillus/metabolism*
Fulltext Kinetics of molybdenum reduction to molybdenum blue by Bacillus sp. strain A.rzi

Othman AR, Bakar NA, Halmi MI, Johari WL, Ahmad SA, Jirangon H, et al.

Biomed Res Int, 2013;2013:371058.
PMID: 24369531 DOI: 10.1155/2013/371058

Molybdenum is very toxic to agricultural animals. Mo-reducing bacterium can be used to immobilize soluble molybdenum to insoluble forms, reducing its toxicity in the process. In this work the isolation of a novel molybdate-reducing Gram positive bacterium tentatively identified as Bacillus sp. strain A.rzi from a metal-contaminated soil is reported. The cellular reduction of molybdate to molybdenum blue occurred optimally at 4 mM phosphate, using 1% (w/v) glucose, 50 mM molybdate, between 28 and 30 °C and at pH 7.3. The spectrum of the Mo-blue product showed a maximum peak at 865 nm and a shoulder at 700 nm. Inhibitors of bacterial electron transport system (ETS) such as rotenone, sodium azide, antimycin A, and potassium cyanide could not inhibit the molybdenum-reducing activity. At 0.1 mM, mercury, copper, cadmium, arsenic, lead, chromium, cobalt, and zinc showed strong inhibition on molybdate reduction by crude enzyme. The best model that fitted the experimental data well was Luong followed by Haldane and Monod. The calculated value for Luong's constants p max, K(s), S(m), and n was 5.88 μmole Mo-blue hr(-1), 70.36 mM, 108.22 mM, and 0.74, respectively. The characteristics of this bacterium make it an ideal tool for bioremediation of molybdenum pollution.

Matched MeSH terms: Bacillus/metabolism*
Molecular characterization, modeling and docking of CYP107CB2 from Bacillus lehensis G1, an alkaliphile

Ang SS, Salleh AB, Chor AL, Normi YM, Tejo BA, Rahman MB

Comput Biol Chem, 2015 Jun;56:19-29.
PMID: 25766878 DOI: 10.1016/j.compbiolchem.2015.02.015

Cytochrome P450s are a superfamily of heme monooxygenases which catalyze a wide range of biochemical reactions. The reactions involve the introduction of an oxygen atom into an inactivated carbon of a compound which is essential to produce an intermediate of a hydroxylated product. The diversity of chemical reactions catalyzed by cytochrome P450s has led to their increased demand in numerous industrial and biotechnology applications. A recent study showed that a gene sequence encoding a CYP was found in the genome of Bacillus lehensis G1, and this gene shared structural similarity with the bacterial vitamin D hydroxylase (Vdh) from Pseudonocardia autotrophica. The objectives of present study was to mine, for a novel CYP from a new isolate B. lehensis G1 alkaliphile and determine the biological properties and functionalities of CYP in this bacterium. Our study employed the usage of computational methods to search for the novel CYP from CYP structural databases to identify the conserved pattern, functional domain and sequence properties of the uncharacterized CYP from B. lehensis G1. A computational homology model of the protein's structure was generated and a docking analysis was performed to provide useful structural knowledge on the enzyme's possible substrate and their interaction. Sequence analysis indicated that the newly identified CYP, termed CYP107CB2, contained the fingerprint heme binding sequence motif FxxGxxxCxG at position 336-345 as well as other highly conserved motifs characteristic of cytochrome P450 proteins. Using docking studies, we identified Ser-79, Leu-81, Val-231, Val-279, Val-383, Ala-232, Thr-236 and Thr-283 as important active site residues capable of stabilizing interactions with several potential substrates, including vitamin D3, 25-hydroxyvitamin D3 and 1α-hydroxyvitamin D3, in which all substrates docked proximally to the enzyme's heme center. Biochemical analysis indicated that CYP107CB2 is a biologically active protein to produce 1α,25-dihydroxyvitamin D3 from 1α-hydroxyvitamin D3. Based on these results, we conclude that the novel CYP107CB2 identified from B. lehensis G1 is a putative vitamin D hydroxylase which is possibly capable of catalyzing the bioconversion of parental vitamin D3 to calcitriol, or related metabolic products.

Matched MeSH terms: Bacillus/metabolism*
Novel starter cultures to inhibit biogenic amines accumulation during fish sauce fermentation

Zaman MZ, Abu Bakar F, Jinap S, Bakar J

Int J Food Microbiol, 2011 Jan 31;145(1):84-91.
PMID: 21183239 DOI: 10.1016/j.ijfoodmicro.2010.11.031

Bacteria with amine oxidase activity have become a particular interest to reduce biogenic amines concentration in food products such as meat and fish sausages. However, little information is available regarding the application of these bacteria in fish sauce. Hence, our study was aimed to investigate the effect of such starter cultures in reducing biogenic amines accumulation during fish sauce fermentation. Staphylococcus carnosus FS19 and Bacillus amyloliquefaciens FS05 isolated from fish sauce which possess amine oxidase activity were used as starter cultures in this study. Fermentation was held for 120 days at 35 °C. The pH value increased in all samples, while salt concentration remained constant throughout fermentation. Aerobic bacteria count was significantly lower (p < 0.05) in the control than in inoculated samples as a result of starter cultures addition. However, it decreased during fermentation due to the growth inhibition by high salt concentration. Proteolytic bacterial count decreased during fermentation with no significant difference (p > 0.05) among samples. These bacteria hydrolyzed protein in anchovy to produce free amino acid precursors for amines formation by decarboxylase bacteria. The presence of biogenic amines producing bacteria in this study was considered to be indigenous from raw material or contamination during fermentation, since our cultures were negative histamine producers. Amino acid histidine, arginine, lysine and tyrosine concentration decreased at different rates during fermentation as they were converted into their respective amines. In general, biogenic amines concentration namely histamine, putrescine, cadaverine and tyramine increased throughout fermentation. However, their concentrations were markedly higher (p < 0.05) in the control (without starter cultures) as compared to the samples treated with starter cultures. Histamine concentration was reduced by 27.7% and 15.4% by Staphylococcus carnosus FS19 and Bacillus amyloliquefaciens FS05, respectively. Both cultures could also reduce other amines during fermentation. After 120 days of fermentation, the overall biogenic amines concentration was 15.9% and 12.5% less in samples inoculated with Staphylococcus carnosus FS19 and Bacillus amyloliquefaciens FS05, respectively, as compared to control samples. These findings emphasized that application of starter cultures with amines oxidase activity in fish sauce fermentation was found to be effective in reducing biogenic amines accumulation.

Matched MeSH terms: Bacillus/metabolism
Danger lurking in the "unknowns": structure-to-function studies of hypothetical protein Bleg1_2437 from Bacillus lehensis G1 alkaliphile revealed an evolutionary divergent B3 metallo-beta-lactamase

Tan SH, Normi YM, Leow AT, Salleh AB, Murad AM, Mahadi NM, et al.

J. Biochem., 2017 02 01;161(2):167-186.
PMID: 28175318 DOI: 10.1093/jb/mvw058

The effectiveness of β-lactam antibiotics as chemotherapeutic agents to treat bacterial infections is gradually threatened with the emergence of antibiotic resistance mechanism among pathogenic bacteria through the production metallo-β-lactamase (MBL). In this study, we discovered a novel hypothetical protein (HP) termed Bleg1_2437 from the genome of alkaliphilic Bacillus lehensis G1 which exhibited MBL-like properties of B3 subclass; but evolutionary divergent from other circulating B3 MBLs. Domain and sequence analysis of HP Bleg1_2437 revealed that it contains highly conserved Zn2+-binding residues such as H54, H56, D58, H59, H131 and H191, important for catalysis, similar with the subclass B3 of MBL. Built 3-D Bleg1_2437 structure exhibited an αββα sandwich layer similar to the well-conserved global topology of MBL superfamily. Other features include a ceiling and floor in the model which are important for accommodation and orientation of β-lactam antibiotics docked to the protein model showed interactions at varying degrees with residues in the binding pocket of Bleg1_2437. Hydrolysis activity towards several β-lactam antibiotics was proven through an in vitro assay using purified recombinant Bleg1_2437 protein. These findings highlight the presence of a clinically important and evolutionary divergent antibiotics-degrading enzyme within the pools of uncharacterized HPs.

Matched MeSH terms: Bacillus/metabolism
Fulltext Production of Biosurfactant Produced from Used Cooking Oil by Bacillus sp. HIP3 for Heavy Metals Removal

Md Badrul Hisham NH, Ibrahim MF, Ramli N, Abd-Aziz S

Molecules, 2019 Jul 18;24(14).
PMID: 31323813 DOI: 10.3390/molecules24142617

Heavy metals from industrial effluents and sewage contribute to serious water pollution in most developing countries. The constant penetration and contamination of heavy metals into natural water sources may substantially raise the chances of human exposure to these metals through ingestion, inhalation, or skin contact, which could lead to liver damage, cancer, and other severe conditions in the long term. Biosurfactant as an efficient biological surface-active agent may provide an alternative solution for the removal of heavy metals from industrial wastes. Biosurfactants exhibit the properties of reducing surface and interfacial tension, stabilizing emulsions, promoting foaming, high selectivity, and specific activity at extreme temperatures, pH, and salinity, and the ability to be synthesized from renewable resources. This study aimed to produce biosurfactant from renewable feedstock, which is used cooking oil (UCO), by a local isolate, namely Bacillus sp. HIP3 for heavy metals removal. Bacillus sp. HIP3 is a Gram-positive isolate that gave the highest oil displacement area with the lowest surface tension, of 38 mN/m, after 7 days of culturing in mineral salt medium and 2% (v/v) UCO at a temperature of 30 °C and under agitation at 200 rpm. An extraction method, using chloroform:methanol (2:1) as the solvents, gave the highest biosurfactant yield, which was 9.5 g/L. High performance liquid chromatography (HPLC) analysis confirmed that the biosurfactant produced by Bacillus sp. HIP3 consists of a lipopeptide similar to standard surfactin. The biosurfactant was capable of removing 13.57%, 12.71%, 2.91%, 1.68%, and 0.7% of copper, lead, zinc, chromium, and cadmium, respectively, from artificially contaminated water, highlighting its potential for bioremediation.

Matched MeSH terms: Bacillus/metabolism*
Fulltext Biotechnological potential of Bacillus salmalaya 139SI: a novel strain for remediating water polluted with crude oil waste

Ismail S, Dadrasnia A

PLoS One, 2015;10(4):e0120931.
PMID: 25875763 DOI: 10.1371/journal.pone.0120931

Environmental contamination by petroleum hydrocarbons, mainly crude oil waste from refineries, is becoming prevalent worldwide. This study investigates the bioremediation of water contaminated with crude oil waste. Bacillus salamalaya 139SI, a bacterium isolated from a private farm soil in the Kuala Selangor in Malaysia, was found to be a potential degrader of crude oil waste. When a microbial population of 108 CFU ml-1 was used, the 139SI strain degraded 79% and 88% of the total petroleum hydrocarbons after 42 days of incubation in mineral salt media containing 2% and 1% of crude oil waste, respectively, under optimum conditions. In the uninoculated medium containing 1% crude oil waste, 6% was degraded. Relative to the control, the degradation was significantly greater when a bacteria count of 99 × 108 CFU ml-1 was added to the treatments polluted with 1% oil. Thus, this isolated strain is useful for enhancing the biotreatment of oil in wastewater.

Matched MeSH terms: Bacillus/metabolism*
Secretory expression and characterization of a highly Ca2+-activated thermostable L2 lipase

Sabri S, Rahman RN, Leow TC, Basri M, Salleh AB

Protein Expr Purif, 2009 Dec;68(2):161-6.
PMID: 19679187 DOI: 10.1016/j.pep.2009.08.002

Thermostable lipases are important biocatalysts, showing many interesting properties with industrial applications. Previously, a thermophilic Bacillus sp. strain L2 that produces a thermostable lipase was isolated. In this study, the gene encoding for mature thermostable L2 lipase was cloned into a Pichia pastoris expression vector. Under the control of the methanol-inducible alcohol oxidase (AOX) promoter, the recombinant L2 lipase was secreted into the culture medium driven by the Saccharomyces cerevisiae alpha-factor signal sequence. After optimization the maximum recombinant lipase activity achieved in shake flasks was 125 U/ml. The recombinant 44.5 kDa L2 lipase was purified 1.8-fold using affinity chromatography with 63.2% yield and a specific activity of 458.1 U/mg. Its activity was maximal at 70 degrees C and pH 8.0. Lipase activity increased 5-fold in the presence of Ca2+. L2 lipase showed a preference for medium to long chain triacylglycerols (C(10)-C(16)), corn oil, olive oil, soybean oil, and palm oil. Stabilization at high temperature and alkaline pH as well as its broad substrate specificity offer great potential for application in various industries that require high temperature operations.

Matched MeSH terms: Bacillus/metabolism
Fulltext A Sco protein among the hypothetical proteins of Bacillus lehensis G1: Its 3D macromolecular structure and association with Cytochrome C Oxidase

Tan SH, Normi YM, Leow AT, Salleh AB, Karjiban RA, Murad AM, et al.

BMC Struct Biol, 2014 Mar 19;14:11.
PMID: 24641837 DOI: 10.1186/1472-6807-14-11

BACKGROUND: At least a quarter of any complete genome encodes for hypothetical proteins (HPs) which are largely non-similar to other known, well-characterized proteins. Predicting and solving their structures and functions is imperative to aid understanding of any given organism as a complete biological system. The present study highlights the primary effort to classify and cluster 1202 HPs of Bacillus lehensis G1 alkaliphile to serve as a platform to mine and select specific HP(s) to be studied further in greater detail.
RESULTS: All HPs of B. lehensis G1 were grouped according to their predicted functions based on the presence of functional domains in their sequences. From the metal-binding group of HPs of the cluster, an HP termed Bleg1_2507 was discovered to contain a thioredoxin (Trx) domain and highly-conserved metal-binding ligands represented by Cys69, Cys73 and His159, similar to all prokaryotic and eukaryotic Sco proteins. The built 3D structure of Bleg1_2507 showed that it shared the βαβαββ core structure of Trx-like proteins as well as three flanking β-sheets, a 310 -helix at the N-terminus and a hairpin structure unique to Sco proteins. Docking simulations provided an interesting view of Bleg1_2507 in association with its putative cytochrome c oxidase subunit II (COXII) redox partner, Bleg1_2337, where the latter can be seen to hold its partner in an embrace, facilitated by hydrophobic and ionic interactions between the proteins. Although Bleg1_2507 shares relatively low sequence identity (47%) to BsSco, interestingly, the predicted metal-binding residues of Bleg1_2507 i.e. Cys-69, Cys-73 and His-159 were located at flexible active loops similar to other Sco proteins across biological taxa. This highlights structural conservation of Sco despite their various functions in prokaryotes and eukaryotes.
CONCLUSIONS: We propose that HP Bleg1_2507 is a Sco protein which is able to interact with COXII, its redox partner and therefore, may possess metallochaperone and redox functions similar to other documented bacterial Sco proteins. It is hoped that this scientific effort will help to spur the search for other physiologically relevant proteins among the so-called "orphan" proteins of any given organism.

Matched MeSH terms: Bacillus/metabolism*

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