scholarly journals Identification of catechol dioxygenases of acinetobacter species, with improved ability to degrade crude oil

2019 ◽  
Author(s):  
Israa M.S. AL-Kadmy ◽  
Ahmed Suhail ◽  
Suhad Abbas Abid ◽  
Sarah Naji Aziz ◽  
Al-Maamoon H. Abed ◽  
...  
Keyword(s):  
Soil Sample ◽  
Crude Extract ◽  
Maximum Growth ◽  
Maximum Activity ◽  
Engine Oil ◽  
Gravimetric Analysis ◽  
Acinetobacter Species ◽  
Spectrometry Analysis ◽  
Biological Domain ◽  
Catechol Dioxygenases

Abstract Background: Various microbes involved in alkanes degradation or reducing organics oil products have been identified among. The goal of this work was to screen the soil bacteria with potential ability to degrade alkanes and characterized using a combination of genetic and physiological methods.Methods: The microbial species of soil sample was isolated and the loss of hydrocarbon was calculated periodically by gravimetric analysis. Isolated bacterial strain with potential ability to degrade the engine oil in Baghdad city was further tested for hydrocarbon-degrading abilities using 2, 6-DCPIP. Colonies of bacteria were counted using serial dilution methods of the soil sample. One of the highly bioactive degrading strains ISN53 was selected and a different fraction of crude extract were analyzed for their hydrocarbon degrading activity. The purified active fraction characterised in LC/MS and SDS PAGE. Presence of chlorocatechol dioxygenases was checked by PCR and expression was analyzed in RT PCR. The envelope of ISn21 isolate was checked for any changes under different conditions using microbial adhesion to the hydrocarbon test (MATH). Results: Serratia marcesens (Sm53) and Acinetobacter venetianus (ISn21) were isolated and ISn21 showed maximum growth on the third day and up to 72% degradation on the 7th day of incubation. LC-MS/MS spectrometry analysis identified the bioactive enzyme catechol dioxygenases in crude extract. The purified catechol dioxygenases enzyme had 35 kDa weight which showed the maximum activity between 48 to 72 hour of incubation. The expression of its gene was 2 fold more than control the maximum expression was observed at 60 to 72 hours of incubation. ISn21 was extremely hydrophobic both in MMV and in LB medium.Conclusion: Using a naturally available resources would be better with efficiency at degrading diesel. Use of like microorganisms during bioremediation operation might supply worthy advances in this important biotechnological/ biological domain.

scholarly journals Identification of catechol dioxygenases of acinetobacter species, with improved ability to degrade crude oil

2019 ◽  
Author(s):  
Israa M.S. AL-Kadmy ◽  
Ahmed Suhail ◽  
Suhad Abbas Abid ◽  
Sarah Naji Aziz ◽  
Al-Maamoon H. Abed ◽  
...  
Keyword(s):  
Soil Sample ◽  
Crude Extract ◽  
Maximum Growth ◽  
Maximum Activity ◽  
Engine Oil ◽  
Gravimetric Analysis ◽  
Acinetobacter Species ◽  
Spectrometry Analysis ◽  
Biological Domain ◽  
Catechol Dioxygenases

Abstract Background: Various microbes involved in alkanes degradation or reducing organics oil products have been identified among. The goal of this work was to screen the soil bacteria with potential ability to degrade alkanes and characterized using a combination of genetic and physiological methods.Methods: The microbial species of soil sample was isolated and the loss of hydrocarbon was calculated periodically by gravimetric analysis. Isolated bacterial strain with potential ability to degrade the engine oil in Baghdad city was further tested for hydrocarbon-degrading abilities using 2, 6-DCPIP. Colonies of bacteria were counted using serial dilution methods of the soil sample. One of the highly bioactive degrading strains ISN53 was selected and a different fraction of crude extract were analyzed for their hydrocarbon degrading activity. The purified active fraction characterised in LC/MS and SDS PAGE. Presence of chlorocatechol dioxygenases was checked by PCR and expression was analyzed in RT PCR. The envelope of ISn21 isolate was checked for any changes under different conditions using microbial adhesion to the hydrocarbon test (MATH). Results: Serratia marcesens (Sm53) and Acinetobacter venetianus (ISn21) were isolated and ISn21 showed maximum growth on the third day and up to 72% degradation on the 7th day of incubation. LC-MS/MS spectrometry analysis identified the bioactive enzyme catechol dioxygenases in crude extract. The purified catechol dioxygenases enzyme had 35 kDa weight which showed the maximum activity between 48 to 72 hour of incubation. The expression of its gene was 2 fold more than control the maximum expression was observed at 60 to 72 hours of incubation. ISn21 was extremely hydrophobic both in MMV and in LB medium.Conclusion: Using a naturally available resources would be better with efficiency at degrading diesel. Use of like microorganisms during bioremediation operation might supply worthy advances in this important biotechnological/ biological domain.

Purification, Properties and Mass Spectrometry Analysis of Two Novel Thermotolerant Endoglucanases from Bacillus akibai I-2

2011 ◽  
Vol 393-395 ◽  
pp. 911-915
Author(s):  
Sen Lin Liu ◽  
Miao Xing
Keyword(s):  
Mass Spectrometry ◽  
Soda Lakes ◽  
Maximum Activity ◽  
Mass Spectrometry Analysis ◽  
16S Rrna Sequence ◽  
Spectrometry Analysis ◽  
16S Rrna Sequence Analysis ◽  
Bacterium Strain ◽  
Terminal Amino ◽  
Alkaliphilic Bacterium

The alkaliphilic bacterium strain Ⅰ-2, which was isolated from soda lakes, was identified as Bacillus akibai by 16S rRNA sequence analysis and suggested to be a new subspecies of genus Bacillus. Two novel thermotolerant alkaline endoglucanases Ⅰ-2-A and Ⅰ-2-B were produced by this alkaliphilic strain. The purified Ⅰ-2-A and Ⅰ-2-B had molecular mass of approximately 60 and 90 kDa, respectively. The optimum pH of Ⅰ-2-A was about 9.0, while that of Ⅰ-2-B was about 8.0. Both enzymes exhibited maximum activity at around 50 °C and were stable up to 50 °C.The two enzymes were resistant to most metal ions and reagents examined. Mass spectrometry analysis indicated that Ⅰ-2-A was probably different from the endoglucanases reported. Ⅰ-2-B showed homology with those of family A5 endoglucanases but low similarity was found in C-terminal amino acid sequence.

scholarly journals Treponema pallidum 3-Phosphoglycerate Mutase Is a Heat-Labile Enzyme That May Limit the Maximum Growth Temperature for the Spirochete

2001 ◽  
Vol 183 (16) ◽  
pp. 4702-4708 ◽  
Author(s):  
Stéphane Benoit ◽  
James E. Posey ◽  
Matthew R. Chenoweth ◽  
Frank C. Gherardini
Keyword(s):  
Treponema Pallidum ◽  
Maximum Growth ◽  
Biochemical Properties ◽  
Maximum Activity ◽  
Phosphoglycerate Mutase ◽  
Heat Sensitivity ◽  
Dependent Manner ◽  
Gene Encoding ◽  
E Coli ◽  
Key Enzyme

ABSTRACT In the causative agent of syphilis, Treponema pallidum, the gene encoding 3-phosphoglycerate mutase, gpm, is part of a six-gene operon (tro operon) that is regulated by the Mn-dependent repressor TroR. Since substrate-level phosphorylation via the Embden-Meyerhof pathway is the principal way to generate ATP inT. pallidum and Gpm is a key enzyme in this pathway, Mn could exert a regulatory effect on central metabolism in this bacterium. To study this, T. pallidum gpm was cloned, Gpm was purified from Escherichia coli, and antiserum against the recombinant protein was raised. Immunoblots indicated that Gpm was expressed in freshly extracted infective T. pallidum. Enzyme assays indicated that Gpm did not require Mn2+ while 2,3-diphosphoglycerate (DPG) was required for maximum activity. Consistent with these observations, Mn did not copurify with Gpm. The purified Gpm was stable for more than 4 h at 25°C, retained only 50% activity after incubation for 20 min at 34°C or 10 min at 37°C, and was completely inactive after 10 min at 42°C. The temperature effect was attenuated when 1 mM DPG was added to the assay mixture. The recombinant Gpm from pSLB2 complemented E. coli strain PL225 (gpm) and restored growth on minimal glucose medium in a temperature-dependent manner. Increasing the temperature of cultures of E. coli PL225 harboring pSLB2 from 34 to 42°C resulted in a 7- to 11-h period in which no growth occurred (compared to wild-type E. coli). These data suggest that biochemical properties of Gpm could be one contributing factor to the heat sensitivity of T. pallidum.

Gene cloning and characterization of thiourocanate hydratase from Burkholderia sp. HME13

2019 ◽  
Vol 167 (3) ◽  
pp. 333-341
Author(s):  
Hisashi Muramatsu ◽  
Haruna Miyaoku ◽  
Syuya Kurita ◽  
Hidenori Matsuo ◽  
Takehiro Kashiwagi ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Amino Acid ◽  
Amino Acid Sequence ◽  
Crude Extract ◽  
Maximum Activity ◽  
Luria Bertani ◽  
Sole Nitrogen Source ◽  
Gene Encoding ◽  
Gene Cloning And Characterization

Abstract A novel enzyme, thiourocanate hydratase, which catalyses the conversion of thiourocanic acid to 3-(5-oxo-2-thioxoimidazolidin-4-yl) propionic acid, was isolated from the ergothioneine-utilizing strain, Burkholderia sp. HME13. When the HME13 cells were cultured in medium containing ergothioneine as the sole nitrogen source, thiourocanate-metabolizing activity was detected in the crude extract from the cells. However, activity was not detected in the crude extract from HME13 cells that were cultured in Luria-Bertani medium. The gene encoding thiourocanate hydratase was cloned and expressed in Escherichia coli, and the recombinant enzyme was purified to homogeneity. The enzyme showed maximum activity at pH 7.5 and 55°C and was stable between pH 5.0 and 10.5, and at temperatures up to 45°C. The Km and Vmax values of thiourocanate hydratase towards thiourocanic acid were 30 μM and 7.1 μmol/min/mg, respectively. The enzyme was strongly inhibited by CuCl2 and HgCl2. The amino acid sequence of the enzyme showed 46% identity to urocanase from Pseudomonas putida, but thiourocanate hydratase had no urocanase activity.

scholarly journals In Vitro α-Amylase and α-Glucosidase Inhibitory and Antioxidant Activities of the Crude Extract and Solvent Fractions of Hagenia abyssinica Leaves

2021 ◽  
Vol 2021 ◽  
pp. 1-9
Author(s):  
Zemene Demelash Kifle ◽  
Simachew Gidey Debeb ◽  
Yaschilal Muche Belayneh
Keyword(s):  
Ethyl Acetate ◽  
Crude Extract ◽  
Antioxidant Activities ◽  
Ethyl Acetate Fraction ◽  
Maximum Activity ◽  
Chloroform Fraction ◽  
Aqueous Fraction ◽  
Water Fraction ◽  
Hagenia Abyssinica

Background. The leaves of Hagenia abyssinica have been used in the management of diabetes mellitus in Ethiopian folk medicine. Thus, this study is aimed at investigating the in vitro α-amylase and α-glucosidase inhibitory and antioxidant activities of the crude extract and solvent fractions of H. abyssinica leaves. Methods. The in vitro α-amylase and α-glucosidase inhibitory and antioxidant activities of the plant extract were assessed using 3,5-dinitrosalicylic acid (DNSA), p-nitro-phenyl-a-D glucopyranoside (p-NPG), and 1,1-diphenyl-2-picrylhydrazyl (DPPH) assays, respectively. Each value of percent inhibition of α-amylase, α-glucosidase, and DPPH scavenging effect was presented as means ± SEM ( n = 3 ). Results. The α-amylase inhibitory activity of the crude extract and solvent fractions was found to be concentration-dependent. The strongest activity was exhibited by the crude extract at the highest concentration with a percentage inhibition of 74.52% (IC50, 14.52 μg/ml) followed by water fraction 68.24% (IC50, 16.31 μg/ml), ethyl acetate fraction 61.57% (IC50, 18.73 μg/ml), and chloroform fraction 56.87% (IC50, 21.57 μg/ml) of H. abyssinica leaves. In the α-glucosidase inhibition assay, the maximum activity was exhibited by the aqueous fraction 62.54% (IC50, 11.67 μg/ml) followed by ethyl acetate fraction 54.97% (IC50, 15.89 μg/ml), crude extract 46.79% (IC50, >16.5 μg/ml), and chloroform fraction 36.44% (IC50, >16.5 μg/ml). In the antioxidant assay, the crude extract exhibited the highest antioxidant activity 86.36% (IC50, 10.25 μg/ml) followed by water fraction 78.59% (IC50, 13.86 μg/ml), ethyl acetate fraction 71.58% (IC50, 16.34 μg/ml), and chloroform fraction 63.65% (IC50, 18.83 μg/ml). Conclusion. This study has revealed that H. abyssinica leaves possess noticeable in vitro α-amylase and α-glucosidase inhibitory and antioxidant activities.

scholarly journals Antimicrobial activities of crude solvent extract and synthesized silver nanoparticles of Opuntia ficus-indica L. Cladodes

2020 ◽  
Vol 49 (4) ◽  
pp. 1085-1093
Author(s):  
IF Bahis ◽  
Mahmoud Moustafa ◽  
Hussein Al-Wadi
Keyword(s):  
Silver Nanoparticles ◽  
Crude Extract ◽  
Klebsiella Oxytoca ◽  
Antimicrobial Activities ◽  
Maximum Activity ◽  
Moderate Activity ◽  
Ag Nps ◽  
Opuntia Ficus Indica ◽  
Maximum Inhibition ◽  
Significant Difference

Opuntia ficus-indica cladodes was examined for their antimicrobial activities by using crude extract and synthesized silver nanoparticles against human pathogenic microbial strains, namely Staphylococcus aureus, Klebsiella pneumoniae, Klebsiella oxytoca, Proteus mirabilis and Candida albicans. All extracts demonstrated moderate activity against the tested microbes in a range between 0.53 and 2.33 cm. The maximum inhibition activity was found to be against K. pneumoniae while the lowest against S. aureus. Synthesized silver nanoparticles (Ag-NPs) showed inhibition activities between 1.82 and 2.03 cm. The maximum activity was recorded against K. oxytoca, while the lowest activities against P. mirabilis. In context of antimicrobial activity, there was no significant difference either solvent crude extract or synthesized silver nanoparticles against tested microbes. Therefore, crude extracts of O. ficus-indica cladodes or Ag-NPs could be used as an alternative natural drug.

scholarly journals Effectiveness of a Lytic Phage SRG1 against Vancomycin-ResistantEnterococcus faecalisin Compost and Soil

2017 ◽  
Vol 2017 ◽  
pp. 1-8 ◽  
Author(s):  
Sidra Rahmat Ullah ◽  
Saadia Andleeb ◽  
Taskeen Raza ◽  
Muhsin Jamal ◽  
Khalid Mehmood
Keyword(s):  
Soil Sample ◽  
Burst Size ◽  
Alternative Therapy ◽  
Maximum Growth ◽  
Sewage Water ◽  
Infection Process ◽  
Lytic Phage ◽  
Lytic Bacteriophage ◽  
Bacteriophage Therapy ◽  
Tract Infections

Nosocomial infections caused by vancomycin-resistantEnterococcushave become a major problem. Bacteriophage therapy is proposed as a potential alternative therapy. Bacteriophages are viruses that infect bacteria and are ubiquitous in nature. Lytic bacteriophage was isolated from sewage water that infects VREF, the causative agent of endocarditis, bacteraemia, and urinary tract infections (UTIs). The phage produced clear plaques with unique clear morphology and well-defined boundaries. TEM results of phage revealed it to be108±0.2 nm long and90±0.5 nm wide. The characterization of bacteriophage revealed that infection process of phage was calcium and magnesium dependent and phage titers were highest under optimum conditions for VREF, with an optimal temperature range of 37–50°C. The maximum growth was observed at 37°C, hence having 100% viability. The latent period for phage was small with a burst size of 512 viral particles per bacterial cell. The phage was tested against various clinical strains and results proved it to be host specific. It can be used as a potential therapeutic agent for VREF infections. The phage efficiently eradicated VREF inoculated in cattle compost, poultry compost, and a soil sample which makes it a potential agent for clearing compost and soil sample.

scholarly journals Influence of pH, Temperature and Various Heavy Metals on β-galactosidase Activity in the Crude Extract of Pleurotus ostreatus

2017 ◽  
Vol 5 (1) ◽  
pp. 29
Author(s):  
Tariq H. Shloul ◽  
Omar M. Atrooz ◽  
Mohammad H. Abukhalil
Keyword(s):  
Heavy Metals ◽  
Crude Extract ◽  
Pleurotus Ostreatus ◽  
Competitive Inhibition ◽  
Maximum Activity ◽  
Galactosidase Activity ◽  
Crude Extracts ◽  
Influence Of Ph ◽  
Hydrolysis Of

The aim of this study was to determine the activity of β-galactosidase in the crude extracts of Pleurotus ostreatus in the presence and absence of various heavy metals. β-galactosidase (EC 3.2.1.23), is a hydrolase enzyme which helps in the hydrolysis of lactose into monosaccharides. Characterization of β-galactosidase from Pleurotus ostreatus was achieved using the substrate 2-nitrophenyl β-D-galactopyranoside (ONPG). The pH and temperature profiles of β-galactosidase showed maximum activity at pH 3.0 and at 50°C, respectively. The Vmax  and Km values of β-galactosidase using ONPG as a substrate was found to be 0.571 μmol/min and 0.307 mM, respectively. These results revealed that the β-galactosidase activity in the crude extracts of Pleurotus ostreatus was changed in the presence of different heavy metals. The results indicated that Hg2+ and Mo2+ have an uncompetitive inhibition on the β- galactosidase activity in the extract of Pleurotus ostreatus by decreasing both Km and Vmax values.  while Al3+, Cu2+, Cr3+, Zn2+ and Ni2+  showed mixed inhibition  activity  by decreasing Vmax values and  by increasing Km values. However, Pb2+ was found to act as a non-competitive inhibition by decreasing Vmax value. The findings suggested that crude extract of Pleurotus ostreatus can be used as a source of β-galactosidase for medical and industrial purposes.

Using Lubricating Oil Filter Debris Analysis to Monitor Abnormal Wear of Aero-Engine

2011 ◽  
Vol 86 ◽  
pp. 821-824 ◽  
Author(s):  
Zi Xin Zhu ◽  
Jun Zhong Zheng ◽  
Dong Chen
Keyword(s):  
Health Management ◽  
Engine Oil ◽  
Lubricating Oil ◽  
Emission Spectrometry ◽  
Composition Analysis ◽  
Distribution Analysis ◽  
Rotating Disc ◽  
Spectrometry Analysis ◽  
Aero Engine ◽  
Wear Modes

With the installation of finer oil filters in the lubrication system of aero-engine, the traditional wear analysis techniques such as Rotating Disc Electrode Atomic Emission Spectrometry Analysis (RDE-AES) become less efficient at detecting abnormal wear failure caused by large debris. These filters capture large debris containing large amounts of tribological information about the operation of an aero-engine. Removal and analysis of the filter debris has proved to be an effective tool for aero-engine health management by determining wear modes and observing failure progression providing long lead times for maintenance. An automated aero-engine oil filter debris analysis system was recently developed. This instrument could efficiently complete filter core washing, debris amount and size distribution analysis, and prepare a patch of the debris for further composition analysis.

scholarly journals ISOLATION AND CHARACTERIZATION OF LIPOLYTIC BACTERIA FROM OIL CONTAMINATED SOIL FROM PETROL BUNK AT SOUTHEAST BANGALORE.

2019 ◽  
Vol 7 (2) ◽  
pp. 30-34
Author(s):  
Ajay Kumar Sahu ◽  
Rahul Nemani ◽  
Prangya Prangya Acharya ◽  
Rupali Sinha ◽  
Subhranil Sengupta ◽  
...  
Keyword(s):  
Crude Oil ◽  
Soil Sample ◽  
Contaminated Soil ◽  
Bacterial Species ◽  
Carbon Sources ◽  
Lubricating Oil ◽  
Contaminated Sites ◽  
Gravimetric Analysis ◽  
Morphological Studies ◽  
Isolation And Characterization

The isolation of oil contaminated sites and gravimetric analysis of degradation in which, two bacterial formed maximum clearing zones on minerals salt medium. An increase in cell number indicating that the bacterial isolation was responsible for the oil degradation .the collected oil contaminated sites at kormongala, Indra nagar, MTTC culture , micrococcus spp., Bacillus spp., pseudomonas spp., which are able to utilize the oil in soil as carbon sources, were added to oil contaminated soil sample , the growth profiles were determined by monitoring the optical density, dry weight and Ph of the culture utilizing lubricating oil as sole sources of carbon, Bushnell  Haas media supplemented with petrol, kerosene and diesel as sole carbon sources was used for isolation of bacteria capable of degrading these petroleum fractionates. From three soil sample and two water sample, a total of nine bacterial strains were isolated capable of degrading petrol, kerosene and diesel with varying tolerance capacities, the isolates were identify by using standard biochemical test and morphological studies and it was determined that these strains belong to six bacterial genera .the present study suggest that the isolated bacterial species could be employed for bioremediation in environment polluted with petroleum and its products, indigenously from the soil and water contaminated with crude oil in the vicinity of oil drilling well were found to be most efficient crude oil utilize as turbidity observed by spectrophotometrically. In the various study of lipolytic bacteria concluded that the taken of oil contaminated soil from from petroleum bunk and to identify their biochemical characterization by using various sources. How it’s helpful for characterize by using of lipolytic bacteria

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