Cover Image

Purification and characterization of recombinant thermostable T1 lipase expressed from Pichia pastoris

Thermostable lipases have high demand in various industries owing to its interesting features. Thermostable T1 lipase gene of Geobacillus zalihae was successfully cloned in Pichia pastoris. However, the characterization of purified lipase is not performed yet. Therefore, this study was conducted to...

Full description

Saved in:
Bibliographic Details
Main Author: Periyasamy, Nalini
Format: Project Paper Report
Language:English
Published: 2015
Online Access:http://psasir.upm.edu.my/id/eprint/78232/1/FBSB%202015%2061%20-%20IR.pdf
http://psasir.upm.edu.my/id/eprint/78232/
Tags: Add Tag
No Tags, Be the first to tag this record!
id my.upm.eprints.78232
record_format eprints
spelling my.upm.eprints.782322020-06-26T07:13:21Z http://psasir.upm.edu.my/id/eprint/78232/ Purification and characterization of recombinant thermostable T1 lipase expressed from Pichia pastoris Periyasamy, Nalini Thermostable lipases have high demand in various industries owing to its interesting features. Thermostable T1 lipase gene of Geobacillus zalihae was successfully cloned in Pichia pastoris. However, the characterization of purified lipase is not performed yet. Therefore, this study was conducted to determine the characteristic of the purified T1 lipase expressed in P. pastoris. The pH of the supernatant was adjusted to pH 7.4 using 1.5 M Tris-HCl and subjected to purification using Nickel-Sepharose resin. The recombinant T1 lipase was purified with 68.6 % recovery and 8.47 fold. The molecular weight of recombinant T1 lipase obtained was approximately 44 kDa. This enzyme exhibited maximum activity at temperature 70 °C and was found to be stable for two hours at 60 °C and 65 °C with relative activity of 80 % and 50 %, respectively. The optimum pH of recombinant T1 lipase was at pH 9 in 0.05 M Tris-HCl. The enzyme was found to be stable at pH 8 and pH 9 with 0.05 M Tris-HCl as its buffer. Finally, the enzyme was found to have broad substrate specificity for natural oils especially with coconut oil with 172 % relative activity. In conclusion, the recombinant T1 lipase expressed in P. pastoris was purified with the highest purification fold and its important characteristics were fully characterized. 2015-06 Project Paper Report NonPeerReviewed text en http://psasir.upm.edu.my/id/eprint/78232/1/FBSB%202015%2061%20-%20IR.pdf Periyasamy, Nalini (2015) Purification and characterization of recombinant thermostable T1 lipase expressed from Pichia pastoris. [Project Paper Report]
institution Universiti Putra Malaysia
building UPM Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Putra Malaysia
content_source UPM Institutional Repository
url_provider http://psasir.upm.edu.my/
language English
description Thermostable lipases have high demand in various industries owing to its interesting features. Thermostable T1 lipase gene of Geobacillus zalihae was successfully cloned in Pichia pastoris. However, the characterization of purified lipase is not performed yet. Therefore, this study was conducted to determine the characteristic of the purified T1 lipase expressed in P. pastoris. The pH of the supernatant was adjusted to pH 7.4 using 1.5 M Tris-HCl and subjected to purification using Nickel-Sepharose resin. The recombinant T1 lipase was purified with 68.6 % recovery and 8.47 fold. The molecular weight of recombinant T1 lipase obtained was approximately 44 kDa. This enzyme exhibited maximum activity at temperature 70 °C and was found to be stable for two hours at 60 °C and 65 °C with relative activity of 80 % and 50 %, respectively. The optimum pH of recombinant T1 lipase was at pH 9 in 0.05 M Tris-HCl. The enzyme was found to be stable at pH 8 and pH 9 with 0.05 M Tris-HCl as its buffer. Finally, the enzyme was found to have broad substrate specificity for natural oils especially with coconut oil with 172 % relative activity. In conclusion, the recombinant T1 lipase expressed in P. pastoris was purified with the highest purification fold and its important characteristics were fully characterized.
format Project Paper Report
author Periyasamy, Nalini
spellingShingle Periyasamy, Nalini
Purification and characterization of recombinant thermostable T1 lipase expressed from Pichia pastoris
author_facet Periyasamy, Nalini
author_sort Periyasamy, Nalini
title Purification and characterization of recombinant thermostable T1 lipase expressed from Pichia pastoris
title_short Purification and characterization of recombinant thermostable T1 lipase expressed from Pichia pastoris
title_full Purification and characterization of recombinant thermostable T1 lipase expressed from Pichia pastoris
title_fullStr Purification and characterization of recombinant thermostable T1 lipase expressed from Pichia pastoris
title_full_unstemmed Purification and characterization of recombinant thermostable T1 lipase expressed from Pichia pastoris
title_sort purification and characterization of recombinant thermostable t1 lipase expressed from pichia pastoris
publishDate 2015
url http://psasir.upm.edu.my/id/eprint/78232/1/FBSB%202015%2061%20-%20IR.pdf
http://psasir.upm.edu.my/id/eprint/78232/
_version_ 1671341110035742720
score 13.211869

Similar Items