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Isolation and characterisation of genes encoding biopolymer manufacturing enzymes
Thesis (MSc)--Stellenbosch University, 2012.
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| Format: | Thesis |
| Language: | en_ZA |
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Stellenbosch : Stellenbosch University
2012
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| _version_ | 1867613964471369728 |
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| access_status_str | Open Access |
| author | Rapp, Telana |
| author2 | Bekker, J. |
| author_browse | Bekker, J. Rapp, Telana |
| author_facet | Bekker, J. Rapp, Telana |
| author_sort | Rapp, Telana |
| collection | Thesis |
| dc_rights_str_mv | Stellenbosch University |
| description | Thesis (MSc)--Stellenbosch University, 2012. |
| format | Thesis |
| id | oai:scholar.sun.ac.za:10019.1/19968 |
| institution | Stellenbosch University (South Africa) |
| language | en_ZA |
| last_indexed | 2026-06-10T12:44:30.757Z |
| license_str | Other — see source repository |
| provenance_str_mv | Harvested via OAI-PMH from SUNScholar — Stellenbosch University Repository |
| publishDate | 2012 |
| publishDateRange | 2012 |
| publishDateSort | 2012 |
| publisher | Stellenbosch : Stellenbosch University |
| publisherStr | Stellenbosch : Stellenbosch University |
| record_format | dspace |
| source_str | SUNScholar — Stellenbosch University Repository |
| spelling | oai:scholar.sun.ac.za:10019.1/19968 Isolation and characterisation of genes encoding biopolymer manufacturing enzymes Rapp, Telana Bekker, J. Kossmann, J. M. Stellenbosch University. Faculty of AgriSciences. Dept. of Genetics. Institute for Plant Biotechnology. Levansucrase Fructan Biosynthesis Dissertations -- Plant biotechnology Theses -- Plant biotechnology Dissertations -- Genetics Theses -- Genetics Biopolymer manufacturing enzymes Biopolymers -- Genetics Thesis (MSc)--Stellenbosch University, 2012. ENGLISH ABSTRACT: Biopolymers exhibit the required material properties to replace conventional, non-biodegradable, petroleum-based polymer products. They have a closed carbon cycle, making them carbon neutral and environmentally friendly. Biopolymers are produced from non-toxic substrates during in vivo enzymatic reactions. Biosynthesis of the most commercially important biopolymers is too complex to be reproduced in in vitro reactions. Identification of the genes responsible for their biosynthesis has been under investigation, with some pathways already elucidated. The genes involved in the biosynthesis of these polymers have been targeted for genetic manipulation to increase productivity, as well as create tailor-made polymers. Novel biopolymers and the genes responsible for their synthesis are of interest for their potential commercial applications. Bacteria produce a wide range of biopolymers and are being implemented as the bio-factories for biopolymer production. They are capable of utilising easily accessible and renewable carbon sources such as sucrose for polymer biosynthesis. Bacteria thus allow for economical production of these environmentally beneficial polymers. In this study, the gene responsible for the production of an unknown biopolymer from an unknown bacterium was identified. The biopolymer producing bacteria were grown on media enriched with sucrose as carbon source, during an expression library screening in a previous study. Expression library technology was used to search for the gene and it was identified as a 424 amino acid levansucrase which had a 100% homology to Leuconostoc mesenteroides M1FT levansucrase (AAT81165.1). Biopolymer analysis revealed that the biopolymer was a levan, a polysaccharide consisting of only fructose molecules with a molecular weight of ± 5 kDa. Analysis of a 516 bp fragment of the 16S rRNA determined that the unknown bacteria were a Pseudomonas species. AFRIKAANSE OPSOMMING: Bio-polimere besit noodsaaklike materiële eienskappe wat toelaat dat dit konvensionele, nie bio-afbreekbare, petroleum-gebasseerde polimeer produkte kan vervang. Hulle het n geslote koolstof kringloop en is dus koolstof neutraal en omgewingsvriendelik. Bio-polimere word vervaardig van nie-toksiese substrate, gedurende ensiematiese reaksies in vivo. Die belangrikste kommersiële bio-polimere se ensiematiese produksie is te kompleks om in ʼn in vitro reaksie te herproduseer. Ondersoeke tot die identifikasie van die gene wat verantwoordelik is vir die produksie van die polimere is onderweg, en sommige produksie paaie is reeds bekend. Die bekende gene word geteiken vir genetiese manipulasie om hulle produktiwiteit te vermeerder en om unieke polimere te produseer. Unieke bio-polimere en die gene wat vir hul produksie verantwoordelik is, is van belang vir hulle potentiële implimentering in komersiële toepassings. Bakteria produseer ʼn verskeidenheid bio-polimere en word as die bio-fabrieke vir polimeerproduksie geimplimenteer. Hulle kan maklik bekombare koolstofbronne, soos sukrose, gebruik om bio-polimere te produseer. Bakteria laat dus die ekonomiese produksie van hierdie omgewingsvriendelike polimere toe. In hierdie studie word die geen wat verantwoordelik is vir die produksie van ʼn onbekende bio-polimeer van ʼn onbekende bakteria, geidentifiseer. Die bakteria was gevind op media, wat verryk was met sukrose as koolstofbron, tydens ʼn vorige studie, waartydens ʼn uitdrukkingsbiblioteek gesif was op hierdie media. Uitdrukkingsbiblioteek tegnologie was gebruik om die geen te vind. Die geen was geidentifiseer as ʼn 424 aminosuur, homo-fruktose-polimeer produseerende geen, ʼn “levansucrase”. Die geen het ʼn 100% homologie met die M1FT “levansucrase” geen (AAT81165.1) van Leuconostoc mesenteroides gehad. Analise van die bio-polimeer het bepaal dat die polimeer ʼn polisakkaried was, wat slegs uit fruktose molekules bestaan het. Die molekulêre gewig van die polimeer was ± 5 kDa. Analise van ʼn 516 bp fragment van die 16S rRNS het bepaal dat die bakteria van die Pseudomonas spesie afkomstig was. 2012-03-16T09:18:54Z 2012-03-30T10:29:24Z 2012-03-16T09:18:54Z 2012-03-30T10:29:24Z 2012-03 Thesis http://hdl.handle.net/10019.1/19968 en_ZA Stellenbosch University 61 p. : ill. application/pdf Stellenbosch : Stellenbosch University |
| spellingShingle | Levansucrase Fructan Biosynthesis Dissertations -- Plant biotechnology Theses -- Plant biotechnology Dissertations -- Genetics Theses -- Genetics Biopolymer manufacturing enzymes Biopolymers -- Genetics Rapp, Telana Isolation and characterisation of genes encoding biopolymer manufacturing enzymes |
| title | Isolation and characterisation of genes encoding biopolymer manufacturing enzymes |
| title_full | Isolation and characterisation of genes encoding biopolymer manufacturing enzymes |
| title_fullStr | Isolation and characterisation of genes encoding biopolymer manufacturing enzymes |
| title_full_unstemmed | Isolation and characterisation of genes encoding biopolymer manufacturing enzymes |
| title_short | Isolation and characterisation of genes encoding biopolymer manufacturing enzymes |
| title_sort | isolation and characterisation of genes encoding biopolymer manufacturing enzymes |
| topic | Levansucrase Fructan Biosynthesis Dissertations -- Plant biotechnology Theses -- Plant biotechnology Dissertations -- Genetics Theses -- Genetics Biopolymer manufacturing enzymes Biopolymers -- Genetics |
| url | http://hdl.handle.net/10019.1/19968 |
| work_keys_str_mv | AT rapptelana isolationandcharacterisationofgenesencodingbiopolymermanufacturingenzymes |