Optimization and in silico analysis of cold-adapted lipase from an antarctic pseudomonas sp strain AMS8 reaction in Triton X-100 reverse micelles
A moderate yield of a purified enzyme can be achieved by using the simple technique of reverse micellar extraction (RME). RME is a liquid–liquid extraction method that uses a surfactant and an organic solvent to extract biomolecules. Instead of traditional chromatographic purification methods, which...
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ftunivpmalaysia:oai:psasir.upm.edu.my:75154 2023-05-15T13:38:42+02:00 Optimization and in silico analysis of cold-adapted lipase from an antarctic pseudomonas sp strain AMS8 reaction in Triton X-100 reverse micelles Abd. Jalil, Fatin Nur Fauzi Ana Raja Abd. Rahman, Raja Noor Zaliha Salleh, Abu Bakar Mohamad Ali, Mohd Shukuri 2018 text http://psasir.upm.edu.my/id/eprint/75154/ http://psasir.upm.edu.my/id/eprint/75154/1/Optimization%20and%20in%20silico%20analysis.pdf en eng MDPI http://psasir.upm.edu.my/id/eprint/75154/1/Optimization%20and%20in%20silico%20analysis.pdf Abd. Jalil, Fatin Nur Fauzi Ana and Raja Abd. Rahman, Raja Noor Zaliha and Salleh, Abu Bakar and Mohamad Ali, Mohd Shukuri (2018) Optimization and in silico analysis of cold-adapted lipase from an antarctic pseudomonas sp strain AMS8 reaction in Triton X-100 reverse micelles. Catalysts, 8 (7). pp. 1-15. ISSN 2073-4344; ESSN: 2073-4344 Article PeerReviewed 2018 ftunivpmalaysia 2020-04-21T15:20:27Z A moderate yield of a purified enzyme can be achieved by using the simple technique of reverse micellar extraction (RME). RME is a liquid–liquid extraction method that uses a surfactant and an organic solvent to extract biomolecules. Instead of traditional chromatographic purification methods, which are tedious and expensive, RME using the nonionic surfactant Triton X-100 and toluene is used as an alternative purification technique to purify a recombinant cold-adapted lipase, AMS8. Various process parameters were optimized to maximize the activity recovery of the AMS8 lipase. The optimal conditions were found to be 50 mM sodium phosphate buffer, pH 7, 0.125 M NaCl, and 0.07 M Triton X-100 in toluene at 10 °C. Approximately 56% of the lipase activity was successfully recovered. Structural analysis of the lipase in a reverse micelle (RM) was performed using an in silico approach. The predicted model of AMS8 lipase was simulated in the Triton X-100/toluene reverse micelles from 5 to 40 °C. The lid 2 was slightly opened at 10 °C. However, the secondary structure of AMS8 was most affected in the non-catalytic domain compared to the catalytic domain, with an increased coil conformation. These results suggest that an AMS8 lipase can be extracted using Triton X-100/water/toluene micelles at low temperature. This RME approach will be an important tool for the downstream processing of recombinant cold-adapted lipases Article in Journal/Newspaper Antarc* Antarctic Universiti Putra Malaysia: PSAS (Perpuskataan Sultan Abuld Samad) Institutional Repository Antarctic Triton ENVELOPE(-55.615,-55.615,49.517,49.517) |
institution |
Open Polar |
collection |
Universiti Putra Malaysia: PSAS (Perpuskataan Sultan Abuld Samad) Institutional Repository |
op_collection_id |
ftunivpmalaysia |
language |
English |
description |
A moderate yield of a purified enzyme can be achieved by using the simple technique of reverse micellar extraction (RME). RME is a liquid–liquid extraction method that uses a surfactant and an organic solvent to extract biomolecules. Instead of traditional chromatographic purification methods, which are tedious and expensive, RME using the nonionic surfactant Triton X-100 and toluene is used as an alternative purification technique to purify a recombinant cold-adapted lipase, AMS8. Various process parameters were optimized to maximize the activity recovery of the AMS8 lipase. The optimal conditions were found to be 50 mM sodium phosphate buffer, pH 7, 0.125 M NaCl, and 0.07 M Triton X-100 in toluene at 10 °C. Approximately 56% of the lipase activity was successfully recovered. Structural analysis of the lipase in a reverse micelle (RM) was performed using an in silico approach. The predicted model of AMS8 lipase was simulated in the Triton X-100/toluene reverse micelles from 5 to 40 °C. The lid 2 was slightly opened at 10 °C. However, the secondary structure of AMS8 was most affected in the non-catalytic domain compared to the catalytic domain, with an increased coil conformation. These results suggest that an AMS8 lipase can be extracted using Triton X-100/water/toluene micelles at low temperature. This RME approach will be an important tool for the downstream processing of recombinant cold-adapted lipases |
format |
Article in Journal/Newspaper |
author |
Abd. Jalil, Fatin Nur Fauzi Ana Raja Abd. Rahman, Raja Noor Zaliha Salleh, Abu Bakar Mohamad Ali, Mohd Shukuri |
spellingShingle |
Abd. Jalil, Fatin Nur Fauzi Ana Raja Abd. Rahman, Raja Noor Zaliha Salleh, Abu Bakar Mohamad Ali, Mohd Shukuri Optimization and in silico analysis of cold-adapted lipase from an antarctic pseudomonas sp strain AMS8 reaction in Triton X-100 reverse micelles |
author_facet |
Abd. Jalil, Fatin Nur Fauzi Ana Raja Abd. Rahman, Raja Noor Zaliha Salleh, Abu Bakar Mohamad Ali, Mohd Shukuri |
author_sort |
Abd. Jalil, Fatin Nur Fauzi Ana |
title |
Optimization and in silico analysis of cold-adapted lipase from an antarctic pseudomonas sp strain AMS8 reaction in Triton X-100 reverse micelles |
title_short |
Optimization and in silico analysis of cold-adapted lipase from an antarctic pseudomonas sp strain AMS8 reaction in Triton X-100 reverse micelles |
title_full |
Optimization and in silico analysis of cold-adapted lipase from an antarctic pseudomonas sp strain AMS8 reaction in Triton X-100 reverse micelles |
title_fullStr |
Optimization and in silico analysis of cold-adapted lipase from an antarctic pseudomonas sp strain AMS8 reaction in Triton X-100 reverse micelles |
title_full_unstemmed |
Optimization and in silico analysis of cold-adapted lipase from an antarctic pseudomonas sp strain AMS8 reaction in Triton X-100 reverse micelles |
title_sort |
optimization and in silico analysis of cold-adapted lipase from an antarctic pseudomonas sp strain ams8 reaction in triton x-100 reverse micelles |
publisher |
MDPI |
publishDate |
2018 |
url |
http://psasir.upm.edu.my/id/eprint/75154/ http://psasir.upm.edu.my/id/eprint/75154/1/Optimization%20and%20in%20silico%20analysis.pdf |
long_lat |
ENVELOPE(-55.615,-55.615,49.517,49.517) |
geographic |
Antarctic Triton |
geographic_facet |
Antarctic Triton |
genre |
Antarc* Antarctic |
genre_facet |
Antarc* Antarctic |
op_relation |
http://psasir.upm.edu.my/id/eprint/75154/1/Optimization%20and%20in%20silico%20analysis.pdf Abd. Jalil, Fatin Nur Fauzi Ana and Raja Abd. Rahman, Raja Noor Zaliha and Salleh, Abu Bakar and Mohamad Ali, Mohd Shukuri (2018) Optimization and in silico analysis of cold-adapted lipase from an antarctic pseudomonas sp strain AMS8 reaction in Triton X-100 reverse micelles. Catalysts, 8 (7). pp. 1-15. ISSN 2073-4344; ESSN: 2073-4344 |
_version_ |
1766110029433274368 |