A rapid two step bacterial DNA extraction method using LasA protease of <em>Pseudomonas aeruginosa</em> MCCB 123
IR@NISCAIR: CSIR-NISCAIR, New Delhi - ONLINE PERIODICALS REPOSITORY (NOPR)
View Archive Info| Field | Value | |
| Title |
A rapid two step bacterial DNA extraction method using LasA protease of <em>Pseudomonas aeruginosa</em> MCCB 123
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| Creator |
Jose, Divya
Jayesh, P Gopinath, Prem Mohandas, A Singh, I S Bright |
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| Subject |
<i>Pseudomonas aeruginosa</i>
LasA protease lytic enzyme <i>Staphylococcus aureus</i> DNA extraction |
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| Description |
495-504
A potent bacteriolytic extracellular protease producing bacterial isolate from coir retting grounds of Kerala, India was identified as <em>Pseudomonas aeruginosa</em> based on phenotypic characteristics and 16S rRNA gene sequence analysis and coded as MCCB 123 (GenBank Accession no. FJ 665510). The enzyme is biocompatible with an IC<sub>50 </sub> of 89.43 ± 3.11µg ml<sup>-1</sup> on mammalian cell line (HeLa). LasA protease was purified to apparent homogeneity with a molecular mass of 20.5 kDa and was found to have a broad range of lytic action on the Gram-positive and Gram-negative bacterial cell walls and also on bacterial consortium. pH, temperature and incubation time for bacterial cell lysis was optimized and found to be 7.0, 35°C and 30 min, respectively with reference to <em>Staphylococcus aureus</em> subsp. <em>aureus</em>. Its broad-spectrum lytic action on wide variety of bacterial cells can be exploited in bacterial DNA extraction without the addition of detergents and chelating agents. This position the enzyme unique over the existing lytic enzymes reported in DNA extraction. This is the first report of <em>P. aeruginosa</em> LasA protease having lytic action on bacterial cell walls other than that of <em>Staphylococcus aureus</em> and its application in rapid extraction of DNA from a wide range of bacteria. |
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| Date |
2018-01-04T04:07:00Z
2018-01-04T04:07:00Z 2017-07 |
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| Type |
Article
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| Identifier |
0975-0967 (Online); 0972-5849 (Print)
http://nopr.niscair.res.in/handle/123456789/43318 |
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| Language |
en_US
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| Rights |
<img src='http://nopr.niscair.res.in/image/cc-license-sml.png'> <a href='http://creativecommons.org/licenses/by-nc-nd/2.5/in' target='_blank'>CC Attribution-Noncommercial-No Derivative Works 2.5 India</a>
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| Publisher |
NISCAIR-CSIR, India
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| Source |
IJBT Vol.16(3) [July 2017]
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