Secretory Production of Recombinant Proteins in Escherichia coli into Vesicles

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Kemiantekniikan korkeakoulu | Bachelor's thesis
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Date

2024-05-19

Department

Major/Subject

Chemical Engineering

Mcode

CHEM3054

Degree programme

Aalto Bachelor's Programme in Science and Technology

Language

en

Pages

38 + 16

Series

Abstract

Recombinant proteins are defined as foreign polypeptides synthesized utilizing biotechnological host organisms for subsequent application in various industrial areas (Nosaki & Miura, 2021). Since production of the first functional polypeptide (i.e., somatostatin) in E. coli in 1977 and the first approval of recombinant insulin therapy in 1982, artificially derived proteins have been applied in pharmaceutics, food production, agriculture, engineering, and research (Clark & Pazdernik, 2016; Pham, 2018). However, despite significant advancements of the field, recombinant protein manufacturing still possesses several limitations related to synthetic capabilities of the host platforms (i.e., yield size, post-translational modifications, and downstream processing), quality of the obtained peptides, and sustainability of the production facilities (Eastwood et al., 2023). Therefore, the current thesis identifies the key advantages and limitations of employing several biotechnological platforms and presents various strategies of E. coli peptide secretion. The paper then develops a novel technique of vesicle-packaged manufacturing of complex proteins devising this enterobacterium, which increases the product yield, and improves sustainability aspects of the process. The experimental part of the work consists of DNA molecular cloning with subsequent gene product purification and expression analysis. The conducted study confirms the promising potential of high yield extracellular secretion of proteins that are either toxic or insoluble if expressed inside of the host bacteria. This directly influences the further affordability of protein-based products, such as pharmaceuticals, addressing several sustainable development goals proposed by the United Nations in 2015 (United Nations, 2023).

Description

Supervisor

Hummel, Michael

Thesis advisor

Aranko, Sesilja

Keywords

escherichia coli, extracellular vesicles, recombinant proteins

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https://urn.fi/URN:NBN:fi:aalto-202406184580

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[kand] Kemian tekniikan korkeakoulu / CHEM