A potential type-II L-asparaginase from marine isolate Bacillus australimaris NJB19: Statistical optimization, in silico analysis and structural modeling

Chakravarty, Namrata; Choudhary, Priyanka; Singh, Jaswant; Singh, Raghuvir

doi:10.1016/j.ijbiomac.2021.01.130

Cited by 18 publications

(10 citation statements)

References 64 publications

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“…De forma similar, Serratia marcescens NCIM 2919 mejoró su producción a pH 7.0, y bacterias aisladas de suelos de Solapur (India) produjeron hasta 103.70 U/mL optimizando su fermentación a pH 7.0, luego del cual se evidenció decrecimiento y mínima productividad a partir de pH 10 (Ghosh et al 2013;Shaikh Khaleed & Khobragade 2016). No obstante, B. licheniformis KKU-KH14 y B. velezensis KB-9, aislados del mar Rojo en Arabia Saudita, revelaron máxima producción a pH 6.5, luego del cual disminuyó considerablemente, en tanto B. australimaris NJB19 produjo 37.93 U/mL a pH 6.77 (Alrumman et al 2019;Mostafa et al 2019;Chakravarty et al 2021). Por el contrario, Shakambari et al (2020) y Venil et al (2009, demostraron que el pH no es un factor significativo para la producción en Pseudomonas plecoglossicida RS1 y Serratia marcescens SB08.…”

Section: Resultados Y Discusiónunclassified

Caracterización bioinformática y producción de L-asparaginasa de Bacillus sp. M62 aislado de las salinas de Maras, Cusco, Perú

Calderón‐Toledo

Tapia-Bañez²,

Jiménez-Aliaga³

et al. 2023

Rev peru biol

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El objetivo del estudio fue realizar la caracterización bioinformática, así como optimizar la producción de L-asparaginasa extracelular de Bacillus sp. M62 aislada de las salinas de Maras (Cusco). Para ello, se verificó la producción de L-asparaginasa mediante el viraje del medio M9 modificado con azul de bromofenol 0.0075%, pH 7.4 a 37 °C por 72 h. A la vez, se extrajo el ADN genómico para amplificar los genes ribosómicos 16S y el gen ansA3. La secuencia aminoacídica codificada por el gen ansA3 se predijo mediante análisis bioinformático. La producción de L-asparaginasa intracelular y extracelular se evaluó a diferentes niveles de glucosa, L-asparagina, NaCl y pH en el medio M9 modificado. Adicionalmente, las actividades enzimáticas de L-asparaginasa y L-glutaminasa se determinaron mediante cuantificación del amonio liberado por el método de Nessler. Así, Bacillus sp. M62 produjo el viraje del medio M9 modificado, obtuvo alta similitud y cercanía evolutiva con Bacillus licheniformis, se encontró que el gen ansA3 amplificado codificaba para 319 aa, dentro de la cual se predijo una secuencia patrón del sitio activo (GFVITHGTDTM ) y 15 sitios inmunogénicos. La producción de L-asparaginasa extracelular fue superior a la intracelular, la que se optimizó de 0.37 U/mL (0.24 U/mg) a 2.15 ± 0.39 U/mL (0.63 U/mg). Finalmente, se encontró que Bacillus sp. M62 presenta L-asparaginasa extracelular con mínima actividad de L-glutaminasa.

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Section: Resultados Y Discusiónunclassified

Caracterización bioinformática y producción de L-asparaginasa de Bacillus sp. M62 aislado de las salinas de Maras, Cusco, Perú

Calderón‐Toledo

Tapia-Bañez²,

Jiménez-Aliaga³

et al. 2023

Rev peru biol

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“…No reuse allowed without permission. The nucleotide sequence encoding for the PrASNase gene obtained in the present study (GenBank accession number MK799854) was translated into the amino acid sequence using the translation tool at the ExPASy online server (https://web.expasy.org/translate/) [26]. The amino acid sequence of PrASNase was compared with the sequence of E. coli asparaginase (EcASNase, Uniprot number P00805), Erwinia chrysanthemi asparaginase (EwASNase, Uniprot number P06608), and Saccharomyces cerevisiae asparaginase (ScASnase, Uniprot number P38986) available in the database using protein data bank (PDB) Blast (https://www.rcsb.org/).…”

Section: Sequence Alignment and Molecular Docking Of Prasnase Genementioning

confidence: 99%

“…The amino acid sequence of PrASNase was compared with the sequence of E. coli asparaginase (EcASNase, Uniprot number P00805), Erwinia chrysanthemi asparaginase (EwASNase, Uniprot number P06608), and Saccharomyces cerevisiae asparaginase (ScASnase, Uniprot number P38986) available in the database using protein data bank (PDB) Blast (https://www.rcsb.org/). The amino acid sequence was analyzed and aligned using Clustal Omega (https://www.ebi.ac.uk/Tools/msa/clustalo/) and Jalview [26][27][28].…”

Section: Sequence Alignment and Molecular Docking Of Prasnase Genementioning

confidence: 99%

Assessment of structural behaviour of new L-asparaginase and SAXS data-based evidence for catalytic activity in its monomeric form

Mihooliya

Nandal

Kalidas

et al. 2023

Preprint

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The present study reports the structural and functional characterization of a new glutaminase-free recombinant L-asparaginase (PrASNase) from Pseudomonas resinovorans IGS-131. PrASNase showed substrate specificity to L-asparagine, and its kinetic parameters, Km, Vmax, and kcat were 9.49x10-3 M, 25.13 IUmL-1min-1, and 3x103 s-1, respectively. The CD spectra showed that PrASNase consists of 30.9% alpha-helix and 69.1% other structures in its native form.FTIR was used for the functional characterization, and molecular docking predicted that the substrate interacts with serine, alanine, and glutamine in the binding pocket of PrASNase. Different from known asparaginases, structural characterization by small-angle X-ray scattering (SAXS) and analytical ultracentrifugation (AUC) unambiguously revealed PrASNase to exist as a monomer in solution at low temperatures and oligomerized to a higher state with temperature rise. Through SAXS studies and enzyme assay, PrASNase was found to be mostly monomer and catalytically active at 37°C. Furthermore, this glutaminase-free PrASNase showed killing effects against WIL2-S with IC50 of 7.4 ug.mL-1 and and TF-1.28 cells with IC50 of 5.6 ug.mL-1. This is probably the first report with significant findings of fully active L-asparaginase in monomeric form using SAXS and AUC and demonstrates the potential of PrASNase in inhibiting cancerous cells, making it a potential therapeutic candidate.

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“…Bacillus australimaris B28A was previously isolated from seawater of costal of Thiruvananthapuram, in with deposit 16SrRNA accession number is MT010836. The strain had an industrial enzymatic activity, with a potential l-asparaginase activity [3] . The B. australimaris B28 genome raw data was comprised a total number of bases sequenced of 1,452,683,420 bps, with total reads in Illumina paired-end sequencing of 9,620,420 and GC content of 42.79%.…”

Section: Data Descriptionmentioning

confidence: 99%

“…The annotated gene using RAST [6] , [7] , [8] categorized identified proteins in twenty-seven main clusters and 318 subsystems of 1396 genes, including asparaginases enzyme. L-asparaginase is recognized as potential anticancer medicine [3] . Subsystems annotation included in (Supplementary 1).…”

Section: Data Descriptionmentioning

confidence: 99%

Whole genome sequence data of Bacillus australimaris strain B28A, isolated from Marine Water in India

Hadi

Edwin²,

Nair

2021

Data in Brief

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Bacillus genus members are dominant in the Eastern Arabian Sea and are known for producing many industrial enzymes. Bacillus australimaris B28A, isolated from seawater, had an enzymatic activity. Here, the whole genome sequence of Bacillus australimaris B28A is reported. The 3,766,107-bp genome, with a GC content of 41.6%, comprised 3936 protein-coding genes, seven ribosomal RNA, and 75 transfer RNA. Several bioactive secondary metabolite genes in the genome, including surfactin, lichenysin, bacillibactin, bacilysin, paenilamicin, fengycin, and carotenoid, were identified using antiSMASH. The 1396 proteins were predicted using RAST, including asparaginase enzyme: an anticancer enzyme. Sequence data have been deposited in the DDBJ/ENA/GenBank database under the accession number JAGQFH000000000. The version described in this paper is JAGQFH000000000.1. The BioProject ID in the GenBank database is PRJNA670955. The raw data is publicly available at " https://www.ncbi.nlm.nih.gov/sra/SRR14203888 ".

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A potential type-II L-asparaginase from marine isolate Bacillus australimaris NJB19: Statistical optimization, in silico analysis and structural modeling

Cited by 18 publications

References 64 publications

Caracterización bioinformática y producción de L-asparaginasa de Bacillus sp. M62 aislado de las salinas de Maras, Cusco, Perú

Caracterización bioinformática y producción de L-asparaginasa de Bacillus sp. M62 aislado de las salinas de Maras, Cusco, Perú

Assessment of structural behaviour of new L-asparaginase and SAXS data-based evidence for catalytic activity in its monomeric form

Whole genome sequence data of Bacillus australimaris strain B28A, isolated from Marine Water in India

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