Purification and characterization of a thermo-halophilic, alkali-stable and extremely benzene tolerant esterase from a thermo-halo tolerant Bacillus cereus strain AGP-03, isolated from ‘Bakreshwar’ hot spring, India

Ghati, Amit; Paul, Goutam

doi:10.1016/j.procbio.2015.01.026

Cited by 25 publications

(22 citation statements)

References 41 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Results also exhibited that the enzymes have proper activities between 20 and 80 ºC. Similar results were also reported for esterases from Picrophilus torridus, 30 Bacillus cereus AGP-03, 26 Pyrococcus furiosus, 31 G. thermodenitrificans T2 25 and Thermomyces lanuginosus. 32 Both pH and temperature optima studies easily showed that mutations performed on TK4PHP templates enhanced these properties as compared with the recombinant enzyme.…”

Section: Effect Of Ph and Temperature On The Esterase Activitiessupporting

confidence: 83%

“…It was reported that the esterases from Geobacillus thermodenitrificans T2 and Bacillus cereus had maximum activity at pH 8.0 and 8.5, with pNPB, respectively. 25,26 Moreover, the esterases from chicken with the optimum activity at pH 8.5 27 and Geobacillus thermoleovorans YN with optimal pH 8.0 have been reported. 28 Temperature influences not only the stability of proteins but also the maximum reaction rate of enzymes by influencing the enzyme and the substrate.…”

Section: Effect Of Ph and Temperature On The Esterase Activitiesmentioning

confidence: 99%

See 1 more Smart Citation

Site-directed mutagenesis and characterization of recombinant phosphotriesterase homology protein from Geobacillus caldoxylosilyticus TK4

Tuncay¹,

Çolak²,

Akatin³

et al. 2019

Rev.Roum.Chim.

View full text Add to dashboard Cite

Many toxic insecticides used worldwide are organophosphates (OPs) derivatives. Phosphotriesterase (PTE) has been rewarding to protect against OP poisoning in vivo or in vitro, associated with advanced catalytic efficiency and stereoselectivity toward the hydrolysis of OPs. Phosphotriesterase homology protein (PHP) exhibits high sequence identity and similarity to PTE. In this study, site-directed mutagenesis on recombinant Geobacillus caldoxylosilyticus TK4 PHP (TK4PHP) was performed for improving the existing esterase activity, even gaining a new PTE activity. After eliminating the deficiencies in the recombinant TK4PHP gene, mutant proteins were purified and characterized biochemically. Considering all the data obtained, it was determined that the major sequence differences between PTE and TK4PHP were removed by three site-directed mutations. However the mutant TK4PHPs did not have PTE activity, it was informed that mutant esterases were more resistance to some metal ions and organic solvents and more thermal stable when it was compared with recombinant type.

show abstract

Section: Effect Of Ph and Temperature On The Esterase Activitiessupporting

confidence: 83%

Section: Effect Of Ph and Temperature On The Esterase Activitiesmentioning

confidence: 99%

Site-directed mutagenesis and characterization of recombinant phosphotriesterase homology protein from Geobacillus caldoxylosilyticus TK4

Tuncay¹,

Çolak²,

Akatin³

et al. 2019

Rev.Roum.Chim.

View full text Add to dashboard Cite

show abstract

“…26 An activity unit was defined as the amount of lipase sufficient to release 1 μmol of p-nitrophenol in 1 min under specific conditions, according to equation 4.…”

Section: Enzymatic Activity Measurementmentioning

confidence: 99%

“…However, it was not possible to maintain this pH, since the change resulted in enzyme precipitation, which can be explained by the fact that this enzyme has a higher activity in alkaline conditions, between pH 6.0 and 10.0 being optimal. 26,44 Therefore, the model solutions for the enzyme application in trichothecene degradation assays were established as phosphate buffer 0.05 mol L -1 at pH 5.0 for laccase and at pH 7.0 for lipase.…”

Section: Enzymatic Activity Standardizationmentioning

confidence: 99%

Evaluation of the Suitability of Analytical Methods in Trichothecene A and B Degradation

Feltrin

Sibaja

Tusnski

et al. 2018

J. Braz. Chem. Soc.

View full text Add to dashboard Cite

An investigation was conducted to standardize conditions for laccase and lipase activity required to maximize trichothecene degradation. The analytical methods were suitable and validated in model solutions for detection of five analytes (T-2 toxin, deoxynivalenol, nivalenol, 3-acetyldeoxynivalenol and 15-acetyldeoxynivalenol), the methods were then applied in the degradation tests of these trichothecenes. The method of salting-out assisted liquid-liquid extraction (SALLE), using three different NaCl concentration levels, obtained recoveries in the range of 47.4-103.4% at pH 5.0 and 36.6-106.8% at pH 7.0, with an intra-day relative standard deviation under 15% for the majority of the compounds. Quantification limits remained in the region of 0.07 μg mL -1 for 15-acetyldeoxynivalenol and 0.3 μg mL -1 for nivalenol. Finally, the suitable analytical methods were applied in a study of trichothecene degradation by enzymatic action, resulting in a reduction of 12.3% for T-2 toxin, 68.4% for deoxynivalenol, 50.2% for 3-acetyldeoxynivalenol and 45.4% for 15-acetyldeoxynivalenol.

show abstract

“…To better characterize such variety, it is useful to use different gene markers able to be detected, also specific functions of present microflora as it was the case of the mcrA gene. In addition, it is possible to explore and describe particular microbial groups or species with promising biotechnological abilities as it was already attempted by others authors (Ghati and Paul 2015;Grivalský et al 2016;Kubo et al 2011). …”

Section: Comparison Of Culture-independent Strategies: High-throughpumentioning

confidence: 99%

Investigation of bacterial and archaeal communities: novel protocols using modern sequencing by Illumina MiSeq and traditional DGGE-cloning

et al. 2016

View full text Add to dashboard Cite

Different protocols based on Illumina high-throughput DNA sequencing and denaturing gradient gel electrophoresis (DGGE)-cloning were developed and applied for investigating hot spring related samples. The study was focused on three target genes: archaeal and bacterial 16S rRNA and mcrA of methanogenic microflora. Shorter read lengths of the currently most popular technology of sequencing by Illumina do not allow analysis of the complete 16S rRNA region, or of longer gene fragments, as was the case of Sanger sequencing. Here, we demonstrate that there is no need for special indexed or tailed primer sets dedicated to short variable regions of 16S rRNA since the presented approach allows the analysis of complete bacterial 16S rRNA amplicons (V1-V9) and longer archaeal 16S rRNA and mcrA sequences. Sample augmented with transposon is represented by a set of approximately 300 bp long fragments that can be easily sequenced by Illumina MiSeq. Furthermore, a low proportion of chimeric sequences was observed. DGGE-cloning based strategies were performed combining semi-nested PCR, DGGE and clone library construction. Comparing both investigation methods, a certain degree of complementarity was observed confirming that the DGGE-cloning approach is not obsolete. Novel protocols were created for several types of laboratories, utilizing the traditional DGGE technique or using the most modern Illumina sequencing.

show abstract

Purification and characterization of a thermo-halophilic, alkali-stable and extremely benzene tolerant esterase from a thermo-halo tolerant Bacillus cereus strain AGP-03, isolated from ‘Bakreshwar’ hot spring, India

Cited by 25 publications

References 41 publications

Site-directed mutagenesis and characterization of recombinant phosphotriesterase homology protein from Geobacillus caldoxylosilyticus TK4

Site-directed mutagenesis and characterization of recombinant phosphotriesterase homology protein from Geobacillus caldoxylosilyticus TK4

Evaluation of the Suitability of Analytical Methods in Trichothecene A and B Degradation

Investigation of bacterial and archaeal communities: novel protocols using modern sequencing by Illumina MiSeq and traditional DGGE-cloning

Contact Info

Product

Resources

About