scholarly journals Thermo-resistant enzyme-producing microorganisms isolated from composting

2023 ◽  
Vol 83 ◽  
Author(s):  
S. P. M. Cotta ◽  
M. S. Marins ◽  
I.E. Marriel ◽  
U.G.P Lana ◽  
E.A. Gomes ◽  
...  
Keyword(s):  
Bacillus Cereus ◽  
Bacillus Licheniformis ◽  
Cellulase Activity ◽  
Ribosomal Gene ◽  
Thermomyces Lanuginosus ◽  
Universal Primers ◽  
Bacillus Species ◽  
Mineral Fertilizers ◽  
Biological Additive

Abstract Organo-mineral fertilizers supplemented with biological additives are an alternative to chemical fertilizers. In this study, thermoresistant microorganisms from composting mass were isolated by two-step procedures. First, samples taken at different time points and temperatures (33 days at 52 ºC, 60 days at 63 ºC, and over 365 days at 26 ºC) were pre-incubated at 80 oC for 30 minutes. Second, the microbial selection by in vitro culture-based methods and heat shock at 60 oC and 100 oC for 2h and 4h. Forty-one isolates were able to grow at 60 °C for 4h; twenty-seven at 100 °C for 2h, and two at 100 °C for 4h. The molecular identification by partial sequencing of the 16S ribosomal gene using universal primers revealed that thirty-five isolates were from eight Bacillus species, one Brevibacillus borstelensis, three Streptomyces thermogriseus, and two fungi (Thermomyces lanuginosus and T. dupontii). Data from amylase, phytase, and cellulase activity assays and the enzymatic index (EI) showed that 38 of 41 thermo-resistant isolates produce at least one enzyme. For amylase activity, the highest EI value was observed in Bacillus licheniformis (isolate 21C2, EI= 4.11), followed by Brevibacillus borstelensis (isolate 6C2, EI= 3.66), Bacillus cereus (isolate 18C2, EI= 3.52), and Bacillus paralicheniformis (isolate 20C2, EI= 3.34). For phytase, the highest EI values were observed for Bacillus cereus (isolate 18C2, EI= 2.30) and Bacillus licheniformis (isolate 3C1, EI= 2.15). Concerning cellulose production, B. altitudinis (isolate 6C1) was the most efficient (EI= 6.40), followed by three Bacillus subtilis (isolates 9C1, 16C2, and 19C2) with EI values of 5.66, 5.84, and 5.88, respectively, and one B. pumilus (isolate 27C2, EI= 5.78). The selected microorganisms are potentially useful as a biological additive in organo-mineral fertilizers and other biotechnological processes.

scholarly journals Thrombolytic Activity of Alkaline Protease Purified from a Mutant Strain Bacillus licheniformis MZK05M9

2018 ◽  
Vol 21 (1) ◽  
pp. 63-70
Author(s):  
Md Asad Uz Zaman ◽  
Taqiyah Akhtar ◽  
ATM Zafrul Azam ◽  
Md Arafat Al Mamun ◽  
Md Mozammel Hoq ◽  
...  
Keyword(s):  
Mutant Strain ◽  
Bacillus Licheniformis ◽  
Serine Proteases ◽  
Specific Activity ◽  
Pharmaceutical Journal ◽  
Crude Enzyme ◽  
Clot Lysis ◽  
Bacillus Species ◽  
Thrombolytic Activity

Investigations were performed to find out new microbial enzymes as thrombolytics having better efficacy and specificity. Mutant strain of Bacillus species, B. licheniformis MZK05M9 was cultured in modified urea-glucose media followed by purification using ammonium sulphate precipitation and ultrafiltration through centricon tube of specific MWCO value. The production method yielded 823.42 units/mg of the crude enzyme from mutant strain MZK05M9 and after purification 37695.64 units/mg. The molecular weight of the purified enzyme was estimated as 27.2 kDa and purification increased its specific activity to 16.5 fold with a recovery of 10%. The purified proteases were identified as serine proteases by irreversible inhibition of activity with phenylmethylsulfonyl fluoride (PMSF) and it exhibited 32.84% thrombolytic activity, by in vitro clot lysis assay. Stability studies showed that crude enzyme from mutant strain MZK05M9 remained stable up to a temperature of 45˚C and showed maximum stability at pH range 7.5 to 8.5. Our observation indicates that proteases produced by Bacillus licheniformis mutant have the potential to be developed as a viable thrombolytic agent.Bangladesh Pharmaceutical Journal 21(1): 63-70, 2018

scholarly journals Isolation, Extraction, Purification, and Molecular Characterization for Thermostable α-Amylase from Locally Isolated Bacillus Species in Sudan

2021 ◽  
Vol 2021 ◽  
pp. 1-8
Author(s):  
Maha A. Rakaz ◽  
Mohammed O. Hussien ◽  
Hanan M. Ibrahim
Keyword(s):  
Bacillus Cereus ◽  
Bacillus Licheniformis ◽  
Soil Bacteria ◽  
Biochemical Characterization ◽  
Amylase Activity ◽  
Industrial Applications ◽  
Soil Samples ◽  
Bacillus Species ◽  
Good Source ◽  
Extraction And Purification

The aim of this study was to isolate some soil bacteria strain that produced α-amylase and subsequent extraction and purification. One hundred soil samples were collected from different geographical areas in Khartoum State such as north Omdurman, Toti Island, and Soba. Samples were analyzed for starch hydrolyzing bacteria. Among several bacteria isolated, Bacillus cereus and Bacillus licheniformis were identified as active α-amylase producers. Both bacteria showed a large zone of clearance of 20 mm when grown on starch-agar plates. The identity was conducted using biochemical characterization and confirmed by sequencing their 16S-rDNA. The constitutive nature of amylase was proved by amplification of the amylase gene from the genome of B. licheniformis. The α-amylase activity from the spent medium of B. cereus and B. licheniformis was optimized at pH 8.0 and temperature of 45°C and 65°C, respectively. The α-amylase produced by both bacteria is alkalophilic and thermophilic. The experiments confirmed that B. licheniformis can be a good source of amylase for industrial applications in Sudan.

scholarly journals Evaluación del crecimiento de cuatro especies del género Bacillus sp., primer paso para entender su efecto biocontrolador sobre Fusarium sp.

Nova ◽  
2016 ◽  
Vol 14 (26) ◽  
pp. 53 ◽  
Author(s):  
Estefania Castañeda Alvarez ◽  
Ligia Consuelo Sánchez
Keyword(s):  
Bacillus Subtilis ◽  
Bacillus Cereus ◽  
Bacillus Licheniformis ◽  
Bacillus Sp ◽  
Fusarium Sp

<p>Evaluar las condiciones de crecimiento de cuatro especies de <em>Bacillus </em>sp. nativas a escala de 10ml en Medio Mínimo de Sales (MMS) como primer paso para entender su acción biocontroladora contra <em>Fusarium </em>sp. <strong>Métodos. </strong>El procedimiento para evaluar el crecimiento de los aislamientos UCMC-TB1, UCMC-TB2, UCMC-TB3 y UCMC-TB4 se realizó utilizando espectrofotometría y recuento directo en placa y pruebas de antagonismo dual en placa para evaluar el efecto controlador contra <em>Fusarium </em>sp. <strong>Resultados. </strong>Se confirmó la identificación por pruebas bioquímicas de los cuatro aislamientos: <em>Bacillus licheniformis, Bacillus subtilis, Bacillus </em><em>pumilus y Bacillus cereus; </em>todas las cepas presentaron antagonismo <em>in vitro</em>. El <em>Bacillus subtilis </em>fue la especie que demostró mayor capacidad antagónica (79,73%PICR) y las características más destacadas de esta cepa fueron su velocidad de crecimiento. El género <em>Bacillus </em>es uno de los más reportados para usar en el control biológico de hongos como <em>Fusarium </em>sp. el cual ataca un gran número de cultivos de interés económico para el sector agrícola en Colombia.</p>

Isolation, identification and in silico study of native cellulase producing bacteria

2019 ◽  
Vol 17 ◽  
Author(s):  
Farzane Kargar ◽  
Mojtaba Mortazavi ◽  
Mahmood Maleki ◽  
Masoud Torkzadeh Mahani ◽  
Younes Ghasemi ◽  
...  
Keyword(s):  
Growth Rate ◽  
Bacillus Cereus ◽  
16S Rdna ◽  
Enzyme Production ◽  
Rational Design ◽  
Cellulase Production ◽  
Bacillus Species ◽  
Bacillus Sp ◽  
Chain Polymer ◽  
Bacillus Toyonensis

Aims: The purpose of this study was to screen the bacteria producing cellulase enzymes and their bioinformatics studies. Background: Cellulose is a long-chain polymer of glucose that hydrolyzes by cellulases to glucose molecules. In order to design the new biotechnological applications, some strategies have been used as increasing the efficiency of enzyme production, generating cost-effective enzymes, producing stable enzymes and identification of new strains. Objective: On the other hand, some bacteria special features have made them suitable candidates for the identification of the new source of enzymes. In this regard, some native strains of bacteria were screened. Method: These bacteria were grown on a culture containing the liquid M9 media containing CMC to ensure the synthesis of cellulase. The formation of a clear area in the culture medium indicated decomposition of cellulose. In the following, the DNA of these bacteria were extracted and their 16S rDNA genes were amplified. Result: The results show that nine samples were able to synthesize cellulase. In following, these strains were identified using 16S rDNA. The results show that these screened bacteria belonged to the Bacillus sp., Alcaligenes sp., Alcaligenes sp., and Enterobacter sp.conclusionThe enzyme activity analysis shows that the Bacillus toyonensis, Bacillus sp. strain XA15-411 Bacillus cereus have produced the maximum yield of cellulases. However, these amounts of enzyme production in these samples are not proportional to their growth rate. As the bacterial growth chart within 4 consecutive days shows that the Alcaligenes sp. Bacillus cereus, Bacillus toyonensis, Bacillus sp. strain XA15-411 have a maximum growth rate. The study of the phylogenetic tree also shows that Bacillus species are more abundant in the production of cellulase enzyme. These bioinformatics analyses show that the Bacillus species have different evolutionary relationships and evolved in different evolutionary time. Other: However, for maximum cellulase production by this bacteria, some information as optimum temperature, optimum pH, carbon and nitrogen sources are needed for the ideal formulation of media composition. The cellulase production is closely controlled in microorganisms and the cellulase yields appear to depend on a variety of factors. However, the further studies are needed for cloning, purification and application of these new microbial cellulases in the different commercial fields as in food, detergent, and pharmaceutical, paper, textile industries and also various chemical industries. However, these novel enzymes can be further engineered through rational design or using random mutagenesis techniques.

scholarly journals Cellulase Activity as a Mechanism for Suppression of Phytophthora Root Rot in Mulches

2011 ◽  
Vol 101 (2) ◽  
pp. 223-230 ◽  
Author(s):  
Brantlee Spakes Richter ◽  
Kelly Ivors ◽  
Wei Shi ◽  
D. M. Benson
Keyword(s):  
Root Rot ◽  
Phytophthora Cinnamomi ◽  
Cellulase Activity ◽  
Enzyme Concentration ◽  
Disease Suppression ◽  
In Vitro Assays ◽  
Infested Soil ◽  
Phytophthora Root Rot ◽  
Standard Curve

Wood-based mulches are used in avocado production and are being tested on Fraser fir for reduction of Phytophthora root rot, caused by Phytophthora cinnamomi. Research with avocado has suggested a role of microbial cellulase enzymes in pathogen suppression through effects on the cellulosic cell walls of Phytophthora. This work was conducted to determine whether cellulase activity could account for disease suppression in mulch systems. A standard curve was developed to correlate cellulase activity in mulches with concentrations of a cellulase product. Based on this curve, cellulase activity in mulch samples was equivalent to a cellulase enzyme concentration of 25 U ml–1 or greater of product. Sustained exposure of P. cinnamomi to cellulase at 10 to 50 U ml–1 significantly reduced sporangia production, but biomass was only reduced with concentrations over 100 U ml–1. In a lupine bioassay, cellulase was applied to infested soil at 100 or 1,000 U ml–1 with three timings. Cellulase activity diminished by 47% between 1 and 15 days after application. Cellulase applied at 100 U ml–1 2 weeks before planting yielded activity of 20.08 μmol glucose equivalents per gram of soil water (GE g–1 aq) at planting, a level equivalent to mulch samples. Cellulase activity at planting ranged from 3.35 to 48.67 μmol GE g–1 aq, but no treatment significantly affected disease progress. Based on in vitro assays, cellulase activity in mulch was sufficient to impair sporangia production of P. cinnamomi, but not always sufficient to impact vegetative biomass.

scholarly journals The growth response of rice (Oryza sativa L. var. FARO 44) in vitro after inoculation with bacterial isolates from a typical ferruginous ultisol

2021 ◽  
Vol 45 (1) ◽  
Author(s):  
Musa Saheed Ibrahim ◽  
Beckley Ikhajiagbe
Keyword(s):  
16S Rrna ◽  
Bacillus Cereus ◽  
Proteus Mirabilis ◽  
Growth Response ◽  
Gene Sequencing ◽  
16S Rrna Gene Sequencing ◽  
Rrna Gene ◽  
Rice Seedlings ◽  
Rrna Gene Sequencing

Abstract Background Rice forms a significant portion of food consumed in most household worldwide. Rice production has been hampered by soil factors such as ferruginousity which has limited phosphorus availability; an important mineral component for the growth and yield of rice. The presence of phosphate-solubilizing bacteria (PSB) in soils has been reported to enhance phosphate availability. In view of this, the present study employed three bacteria species (BCAC2, EMBF2 and BCAF1) that were previously isolated and proved P solubilization capacities as inocula to investigate the growth response of rice germinants in an in vitro setup. The bacteria isolates were first identified using 16S rRNA gene sequencing and then applied as inoculum. The inolula were prepared in three concentrations (10, 7.5 and 5.0 ml) following McFarland standard. Viable rice (var. FARO 44) seeds were sown in petri dishes and then inoculated with the three inocula at the different concentrations. The setup was studied for 28 days. Results 16S rRNA gene sequencing identified the isolates as: isolate BCAC2= Bacillus cereus strain GGBSU-1, isolate BCAF1= Proteus mirabilis strain TL14-1 and isolate EMBF2= Klebsiella variicola strain AUH-KAM-9. Significant improvement in rice germination, morphology, physiology and biomass parameters in the bacteria-inoculated setups was observed compared to the control. Germination percentage after 4 days was 100 % in the inoculated rice germinants compared to 65% in the control (NiS). Similarly, inoculation with the test isolates enhanced water-use efficiency by over 40%. The rice seedlings inoculated with Bacillus cereus strain GGBSU-1 (BiS) showed no signs of chlorosis and necrosis throughout the study period as against those inoculated with Proteus mirabilis strain TL14-1 (PiS) and Klebsiella variicola strain AUH-KAM-9 (KiS). Significant increase in chlorophyll-a, chlorophyll-b and alpha amylase was observed in the rice seedlings inoculated with BiS as against the NiS. Conclusion Inoculating rice seeds with Bacillus cereus strain GGBSU-1, Proteus mirabilis strain TL14-1 and Klebsiella variicola strain AUH-KAM-9 in an in vitro media significantly improved growth parameters of the test plant. Bacillus cereus strain GGBSU-1 showed higher efficiency due to a more improved growth properties observed.

scholarly journals Attenuation of Aeromonas hydrophila Infection in Carassius auratus by YtnP, a N-acyl Homoserine Lactonase from Bacillus licheniformis T-1

Antibiotics ◽  
2021 ◽  
Vol 10 (6) ◽  
pp. 631
Author(s):  
Mengfan Peng ◽  
Wentao Tong ◽  
Zhen Zhao ◽  
Ling Xiao ◽  
Zhaoyue Wang ◽  
...  
Keyword(s):  
Virulence Factors ◽  
Bacillus Licheniformis ◽  
Aeromonas Hydrophila ◽  
Biofilm Formation ◽  
Carassius Auratus ◽  
Quorum Quenching ◽  
Inhibition Rate ◽  
Sequence Identity

In this experiment, the quorum quenching gene ytnP of Bacillus licheniformis T-1 was cloned and expressed, and the effect against infection of Aeromonas hydrophila ATCC 7966 was evaluated in vitro and vivo. The BLAST results revealed a 99% sequence identity between the ytnP gene of T-1 and its homolog in B.subtilis sub sp. BSP1, and the dendroGram showed that the similarity in the YtnP protein in T-1 was 100% in comparison with B.subtilis 3610, which was categorized as the Aidc cluster of the MBL family. The AHL lactonase activity of the purified YtnP was detected as 1.097 ± 0.7 U/mL with C6-HSL as the substrate. Otherwise, purified YtnP protein could significantly inhibit the biofilm formation of A.hydrophila ATCC 7966 with an inhibition rate of 68%. The MIC of thiamphenicol and doxycycline hydrochloride against A. hydrophila reduced from 4 μg/mL and 0.5 μg/mL to 1 μg/mL and 0.125 μg/mL, respectively, in the presence of YtnP. In addition, YtnP significantly inhibited the expression of five virulence factors hem, ahyB, ast, ep, aerA of A. hydrophila ATCC 7966 as well (p < 0.05). The results of inhibition on virulence showed a time-dependence tendency, while the strongest anti-virulence effects were within 4–24 h. In vivo, when the YtnP protein was co-injected intraperitoneally with A. hydrophila ATCC 7966, it attenuated the pathogenicity of A. hydrophila and the accumulated mortality was 27 ± 4.14% at 96 h, which was significantly lower than the average mortality of 78 ± 2.57% of the Carassius auratus injected with 108 CFU/mL of A. hydrophila ATCC 7966 only (p < 0.001). In conclusion, the AHL lactonase in B. licheniformis T-1 was proven to be YtnP protein and could be developed into an agent against infection of A. hydrophila in aquaculture.

In vitro maturation pathway of a glutamyl endopeptidase precursor from Bacillus licheniformis

Biochimie ◽  
2005 ◽  
Vol 87 (6) ◽  
pp. 529-537 ◽  
Author(s):  
L.A. Trachuk ◽  
A.S. Shcheglov ◽  
E.I. Milgotina ◽  
G.G. Chestukhina
Keyword(s):  
Bacillus Licheniformis ◽  
In Vitro Maturation ◽  
Glutamyl Endopeptidase

scholarly journals Bacillus cereus Induces Permeability of an In Vitro Blood-Retina Barrier

2008 ◽  
Vol 76 (4) ◽  
pp. 1358-1367 ◽  
Author(s):  
A. L. Moyer ◽  
R. T. Ramadan ◽  
J. Thurman ◽  
A. Burroughs ◽  
M. C. Callegan
Keyword(s):  
Quorum Sensing ◽  
Bacillus Cereus ◽  
Barrier Function ◽  
Pigment Epithelium ◽  
Retinal Pigment ◽  
Toxin Production ◽  
Global Regulator ◽  
Barrier Permeability ◽  
Cell Monolayers

ABSTRACT Most Bacillus cereus toxin production is controlled by the quorum-sensing-dependent, pleiotropic global regulator plcR, which contributes to the organism's virulence in the eye. The purpose of this study was to analyze the effects of B. cereus infection and plcR-regulated toxins on the barrier function of retinal pigment epithelium (RPE) cells, the primary cells of the blood-retina barrier. Human ARPE-19 cells were apically inoculated with wild-type or quorum-sensing-deficient B. cereus, and cytotoxicity was analyzed. plcR-regulated toxins were not required for B. cereus-induced RPE cytotoxicity, but these toxins did increase the rate of cell death, primarily by necrosis. B. cereus infection of polarized RPE cell monolayers resulted in increased barrier permeability, independent of plcR-regulated toxins. Loss of both occludin and ZO-1 expression occurred by 8 h postinfection, but alterations in tight junctions appeared to precede cytotoxicity. Of the several proinflammatory cytokines analyzed, only interleukin-6 was produced in response to B. cereus infection. These results demonstrate the deleterious effects of B. cereus infection on RPE barrier function and suggest that plcR-regulated toxins may not contribute significantly to RPE barrier permeability during infection.

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