Exploring of bacterial lipoxygenase expression in yeast saccharomyces cerevisiae
Institutional Repository of Vilnius University, 2018
Online
unknown
Zugriff:
Chemical synthesis methods are still widely used in various industrial processes. The production of polyurethanes (PU), which is the third most produced polymer in the world, is not the exception. Currently, the production of PU is directed towards the replacement of chemically based synthesis and oil-based components with environmentally friendly and renewable materials, for example, vegetable oils with expanded functionalities. Therefore, there is a demand need for enzymes which could convert vegetable oils into polyols. One of such is lipoxygenase, which directly catalyzes hydroperoxidation of polyunsaturated fatty acids; however, it is not widely studied, particularly of bacterial origin, and commercially is not available. The aim of the this research was to investigate the bacterial lox gene expression in heterologous Saccharomyces cerevisiae system and to determine the impact of two different types of signaling peptides on secretion of lipoxygenase. Initially, constructed plasmids containing two different signaling peptides (α factor and EPG), were used for the investigation of LOX expression. The analysis revealed that more effective system was the one with the EPG signaling peptide. Despite the fact that the protein was found in all three fractions (supernatant, soluble and insoluble protein fractions), the majority of it was insoluble. Thus, the optimization of induction conditions for the better protein expression was carried out. The decrease of induction temperature closer to the temperature optimum of lipoxygenases, had only slightl influence for the increase of target protein secretion. Additionally, the detailed analysis data indicated that His-tag attached to lox gene encumber the expression of lipoxygenase.
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Exploring of bacterial lipoxygenase expression in yeast saccharomyces cerevisiae
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Autor/in / Beteiligte Person: | Venckus, Žymantas ; Matijošytė, Inga |
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Veröffentlichung: | Institutional Repository of Vilnius University, 2018 |
Medientyp: | unknown |
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