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Optimised bacterial production and characterisation of natural antimicrobial peptides with potential application in agriculture

Thesis (PhD)--Stellenbosch University, 2016

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Main Author: Vosloo, Johan Arnold
Other Authors: Rautenbach, Marina
Format: Thesis
Language:en_ZA
Published: Stellenbosch : Stellenbosch University 2016
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access_status_str Open Access
author Vosloo, Johan Arnold
author2 Rautenbach, Marina
author_browse Rautenbach, Marina
Vosloo, Johan Arnold
author_facet Rautenbach, Marina
Vosloo, Johan Arnold
author_sort Vosloo, Johan Arnold
collection Thesis
dc_rights_str_mv Stellenbosch University
description Thesis (PhD)--Stellenbosch University, 2016
format Thesis
id oai:scholar.sun.ac.za:10019.1/98411
institution Stellenbosch University (South Africa)
language en_ZA
last_indexed 2026-06-10T12:46:44.579Z
license_str Other — see source repository
provenance_str_mv Harvested via OAI-PMH from SUNScholar — Stellenbosch University Repository
publishDate 2016
publishDateRange 2016
publishDateSort 2016
publisher Stellenbosch : Stellenbosch University
publisherStr Stellenbosch : Stellenbosch University
record_format dspace
source_str SUNScholar — Stellenbosch University Repository
spelling oai:scholar.sun.ac.za:10019.1/98411 Optimised bacterial production and characterisation of natural antimicrobial peptides with potential application in agriculture Vosloo, Johan Arnold Rautenbach, Marina Snoep, Jacky L. Stellenbosch University. Faculty of Science. Department Biochemistry Tyrocidine Bacterial production UCTD Bacteria -- Pathogens Antimicrobial peptides (AMPs) Green biocides Thesis (PhD)--Stellenbosch University, 2016 ENGLISH ABSTRACT : The loss incurred due to microbial spoilage is a great concern to the agricultural and food industries. Microbial pathogens have typically been controlled through the use of chemical control agents and antibiotics. However, combination of emerging microbial resistance together with consumer opposition and regulation toward these chemical agents has instigated the need for so called green-biocides which have a reduced ecological impact together with lower potential for resistance development. The tyrocidines and their analogues, cyclic decapeptides produced by the soil bacterium Bacillus aneurinolyticus, show potential to serve as green-biocides. Potent antimicrobial activity toward bacterial and fungal pathogens which affect both the food and agricultural industries is achieved by different cyclodecapeptide analogues. The objective of this study was the development and evaluation of the future potential of the cyclodecapeptides produced by B. aneurinolyticus to serve as green-biocides. In this thesis an overview is given in Chapter 1 of the challenges facing agricultural production due to microbial spoilage and resistance, as well as the potential of antimicrobial peptides, specifically the tyrocidines and analogues to address this problem. The optimal conditions allowing for future large scale production of target-specific subsets of these cyclodecapeptides and economical downstream purification of the peptide mixtures and single peptides is covered in Chapters 2-4. The produced cyclodecapeptide mixtures/formulations were investigated to elucidate some parameters which may allow for optimisation to target different agricultural and/or food related pathogens/problem organisms (Chapters 5 and 6). Initial formulation parameters were evaluated together with the relative safety of their application within an agricultural setting by determining their in vivo toxicity toward honey bees (Chapter 7). A Summary of the findings in each of the different chapters is presented together with proposed future work (Chapter 8). The experimental chapters of this thesis were written as independent units so as to enable future publication. In some chapters repetitions were unavoidable, every attempt was made to keep this to a minimum. The first goal of this study was the increased natural production of these cyclodecapeptides to enabling their tailored application to target different pathogens in an agricultural environment. In order to achieve the goal of this study the following objectives were set: Manipulation of the natural cyclodecapeptide profile of B. aneurinolyticus to produce defined peptide subsets and single peptides. (Chapter 2). Construction of a computational model of cyclodecapeptide production by B. aneurinolyticus (Chapter 3). Optimise production and purification methodology of the tyrocidines and their analogues for future large scale production (Chapter 4). The second goal was to characterise the peptides in order to facilitate formulation of the peptides for in vivo applications. In order to achieve the goal of this study the following objectives were set: Investigate the structure and oligomerisation relationships of the major cyclodecapeptide analogues produced in the different subsets (Chapter 5). Correlation of the oligomerisation characteristics of co-produced cyclodecapeptides with their antimicrobial activity toward two representative target organisms (Chapter 6). Formulation of the natural cyclodecapeptide extracts and investigation of their in vivo toxicity toward honey bees as representative non-target species (Chapter 7). Doctoral 2016-03-09T14:16:00Z 2016-03-09T14:16:00Z 2016-03 Thesis http://hdl.handle.net/10019.1/98411 en_ZA Stellenbosch University xxi, 238 pages : illustrations (some colour) application/pdf Stellenbosch : Stellenbosch University
spellingShingle Tyrocidine
Bacterial production
UCTD
Bacteria -- Pathogens
Antimicrobial peptides (AMPs)
Green biocides
Vosloo, Johan Arnold
Optimised bacterial production and characterisation of natural antimicrobial peptides with potential application in agriculture
title Optimised bacterial production and characterisation of natural antimicrobial peptides with potential application in agriculture
title_full Optimised bacterial production and characterisation of natural antimicrobial peptides with potential application in agriculture
title_fullStr Optimised bacterial production and characterisation of natural antimicrobial peptides with potential application in agriculture
title_full_unstemmed Optimised bacterial production and characterisation of natural antimicrobial peptides with potential application in agriculture
title_short Optimised bacterial production and characterisation of natural antimicrobial peptides with potential application in agriculture
title_sort optimised bacterial production and characterisation of natural antimicrobial peptides with potential application in agriculture
topic Tyrocidine
Bacterial production
UCTD
Bacteria -- Pathogens
Antimicrobial peptides (AMPs)
Green biocides
url http://hdl.handle.net/10019.1/98411
work_keys_str_mv AT vosloojohanarnold optimisedbacterialproductionandcharacterisationofnaturalantimicrobialpeptideswithpotentialapplicationinagriculture