RHAMNOLIPIDS BIOSYNTHESIS IN PSEUDOMONAS AERUGINOSA ONU 301 IN THE PRESENCE OF EXOGENOUS SIGNALING QUINOLONE
Aim: Discovery of the P. aeruginosa ONU 301rhamnolipids biosynthesis and rhamnosyltransferase 2 activity in presence of Pseudomonas aeruginosa exogenous quorum sensing signal molecule 2-heptyl-3-hydroxy-4-quinolon (PQS). Methods. Pseudomonas aeruginosa ONU 302 were cultured in the Giss medium with 2...
Main Authors: | , , |
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Format: | Article |
Language: | English |
Published: |
Odessa I. I. Mechnikov National University
2014-06-01
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Series: | Mìkrobìologìâ ì Bìotehnologìâ |
Subjects: | |
Online Access: | http://mbt.onu.edu.ua/article/view/48255 |
Summary: | Aim: Discovery of the P. aeruginosa ONU 301rhamnolipids biosynthesis and rhamnosyltransferase 2 activity in presence of Pseudomonas aeruginosa exogenous quorum sensing signal molecule 2-heptyl-3-hydroxy-4-quinolon (PQS). Methods. Pseudomonas aeruginosa ONU 302 were cultured in the Giss medium with 2% glucose at 37 оС 24 h. All discoveries were performed in «plancton-biofilm» system with using of the «Nunclon» 48-well plates. Di- and monorhamnolipids separation conducted by TLC methods and its content was determined by orcinol test. Rhamnosyltransferase 2 (RhlC) activity was analysed in P. aeruginosa cell extracts using a rhamnosyltransferase assay specific for the addition of L-rhamnose to monorhamnolipid. Results. After 16 h of growth, there is approximately 4.2 times more biosurfactant in cultures supplemented with PQS compared with the control. After 24 hours its level in culture medium was 16.0 mg/ml in the presence of 80 μМ PQS and 3.7 mg/ml in the absence of PQS. The dirhamnolipids content in control culture after 8 hours was less than the monorhamnolipids. But after 24 hours its level was in 2.86 time higher. The dirhamnolipid/ monorhamnolipid ratio increased 24 hours later in 1.25; 1.55 and 2.25 times in presence of 40, 60 and 80 μМ signaling quinolon concentration, respectively. The additions of PQS at the time of inoculation are sufficient to induce RhlC activity during the transition to stationary phase. So, after eight hours in the presence of 40, 60 or 80 μМ PQS rhamnosyltransferase 2 activity was 1.5, 1.9 and 2.2 times higher as compared with the control. After 24 hours this enzymatic activity was higher at 45%, 85% and 110%, respectively. Conclusion. It is concluded that, Pseudomonas aeruginosa quorum sensing signal molecule 2-heptyl-3-hydroxy-4-quinolon (PQS) can be used in biothechnology to increase the yield of biosurfactants and enrich them with dirhamnolipids |
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ISSN: | 2076-0558 2307-4663 |