Generation of novel random mutagenesis lipase libraries via directed evolution

Yenilmez, Batuhan Orbay (2013) Generation of novel random mutagenesis lipase libraries via directed evolution. [Thesis]

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Abstract

Lipases (triacylglycerol acylhydrolases, EC 3.1.1.3) function as significant biocatalysts in biotechnological applications. The fact that their mechanism, selectivity and structure is well known make lipases a suitable candidate for studies of protein engineering and directed evolution. Using the merits of DNA shuffling method, directed evolution and random mutagenesis,libraries of mutant lipases are constructed with improved features and functionality of pre-existing ones, which in turn encourages the use of industrial lipases in applications such as biosensors, pharmaceuticals, agrochemicals, bioremediation, etc. With increasing demand on lipase production for commercial use, it has thus become crucial to identify and isolate novel and target-specific lipases, as well as optimizing existing ones for acquisition of desired functionality. The aim of this study is to generate mutant lipase libraries using directed evolution and to screen for a candidate biocatalyst. There are two lipases of interest, the mesophilic Aspergillus niger lipase (ANL) and the thermophilic Bacillus thermocatenulatus lipase (BTL), which were shuffled in order to obtain a mutant library that would have the desired features such as increased thermostability, pH stability and a broader range of substrate specificity.
Item Type: Thesis
Uncontrolled Keywords: DNA shuffling. -- Directed evolution. -- Industrial lipases. -- Random mutagenesis. -- DNA karma metodu. -- Yönlendirilmiş evrim. -- Endüstriyel lipazlar. -- Rastgele mutasyon yöntemi.
Subjects: T Technology > TA Engineering (General). Civil engineering (General) > TA164 Bioengineering
Divisions: Faculty of Engineering and Natural Sciences > Academic programs > Biological Sciences & Bio Eng.
Faculty of Engineering and Natural Sciences
Depositing User: IC-Cataloging
Date Deposited: 13 Oct 2015 15:16
Last Modified: 26 Apr 2022 10:05
URI: https://research.sabanciuniv.edu/id/eprint/27312

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