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Fumaric acid production with Rhizopus oryzae : quantifying time-dependent variations in catabolic flux

Dissertation (MEng)--University of Pretoria, 2015.

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Other Authors: Nicol, Willie
Format: Thesis
Language:English
Published: University of Pretoria 2015
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access_status_str Open Access
author2 Nicol, Willie
author_browse Nicol, Willie
author_facet Nicol, Willie
collection Thesis
dc_rights_str_mv © 2015 University of Pretoria. All rights reserved. The copyright in this work vests in the University of Pretoria. No part of this work may be reproduced or transmitted in any form or by any means, without the prior written permission of the University of Pretoria.
description Dissertation (MEng)--University of Pretoria, 2015.
format Thesis
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institution University of Pretoria (South Africa)
language English
last_indexed 2026-06-10T12:38:39.160Z
license_str Other — see source repository
provenance_str_mv Harvested via OAI-PMH from UPSpace — University of Pretoria Institutional Repository
publishDate 2015
publishDateRange 2015
publishDateSort 2015
publisher University of Pretoria
publisherStr University of Pretoria
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source_str UPSpace — University of Pretoria Institutional Repository
spelling oai:repository.up.ac.za:2263/51118 Fumaric acid production with Rhizopus oryzae : quantifying time-dependent variations in catabolic flux Nicol, Willie andre.naude.mail@gmail.com Naude, Andre UCTD Rhizopus oryzae Fumaric acid Metabolic flux analysis DO PH Immobilisation Engineering, built environment and information technology theses SDG-09 Engineering, built environment and information technology theses SDG-12 Engineering, built environment and information technology theses SDG-17 Dissertation (MEng)--University of Pretoria, 2015. A technique for analysing batch fumaric acid fermentations is presented. Off-gas measurements were included in the analysis, which assisted in improving the accuracy by means of data reconciliation due to these extra measurements resulting in an overdetermined mass-balance. Instantaneous rate estimations were obtained through smooth polynomial fits on the consolidated dataset. The primary objective was to use this technique to analyse the metabolic flux distribution as the fermentation progressed. This led to the identification of various metabolic phases that could be exploited for favourable results in future process designs. The secondary objective was to investigate the influence of pH and dissolved oxygen (DO) concentration on the metabolic flux distribution using the described technique, with the focus shifted to differences in instantaneous rate and yield characteristics, rather than accumulative differences. Three distinct metabolic phases were present during this study. Phase A relates to the adjustment period when changing mediums and is characterised by zero fumaric acid production. Phase B started with the commencement of fumaric acid production; the three major metabolic pathways (fumaric acid, ethanol and respiration) were all active during this phase. Most of the fumaric acid was produced during this stage while ethanol production steadily declined. Phase C started when ethanol production reached zero and therefore only the fumaric acid and respiration pathways were active during this phase. Unlike with the ethanol pathway, the activity of the respiration pathway did not decline as the fermentation progressed. The diminishing ethanol production was interpreted as inactivation of the anaerobic part of the biomass while the aerobic part remained unaffected. The respiration capacity (0.8 mmol O2/g biomass.h) was identical for all the DO 60% and 80% fermentations, which suggests that respiration was limited by factors other than the DO. The full respiration capacity was not achieved with limited oxygenation (DO 20%). Instantaneous fumaric acid yields (0.8 g/g for DO 80%) were high during Phase C, but the high ethanol production during Phase B wasted a major fraction of the glucose which led to low overall fumaric acid yields (0.52 g/g for DO 80%). Zero ethanol production and constant respiration imply that the fumaric acid yield depends solely on the fumaric acid production rate during Phase C. It was found that this rate was inhibited by the fumarate concentration itself, resulting in lower rates (and yields) at high fumarate concentrations. High pH and DO levels countered this inhibition effect. Based on the results, it is recommended that fermentations should be operated at a pH of 5 and a DO of 80%, and that most of the fumaric acid should be produced during Phase C. This can be achieved by batch fermentations with higher initial glucose concentrations (> 100 g/L), by fed-batch fermentations with late glucose addition or by continuous fermentations with concentrations and pretreatment conditions similar to those of the start of Phase C. tm2015 mi2026 Chemical Engineering MEng Unrestricted SDG-09: Industry, innovation and infrastructure SDG-12: Responsible consumption and production SDG-17: Partnerships for the goals 2015-12-08T07:34:58Z 2015-12-08T07:34:58Z 2015-09-03 2015 Dissertation Naude, A 2015, Fumaric acid production with Rhizopus oryzae : quantifying time-dependent variations in catabolic flux, MEng Dissertation, University of Pretoria, Pretoria, viewed yymmdd <http://hdl.handle.net/2263/51118> S2015 http://hdl.handle.net/2263/51118 en © 2015 University of Pretoria. All rights reserved. The copyright in this work vests in the University of Pretoria. No part of this work may be reproduced or transmitted in any form or by any means, without the prior written permission of the University of Pretoria. application/pdf University of Pretoria
spellingShingle UCTD
Rhizopus oryzae
Fumaric acid
Metabolic flux analysis
DO
PH
Immobilisation
Engineering, built environment and information technology theses SDG-09
Engineering, built environment and information technology theses SDG-12
Engineering, built environment and information technology theses SDG-17
Fumaric acid production with Rhizopus oryzae : quantifying time-dependent variations in catabolic flux
title Fumaric acid production with Rhizopus oryzae : quantifying time-dependent variations in catabolic flux
title_full Fumaric acid production with Rhizopus oryzae : quantifying time-dependent variations in catabolic flux
title_fullStr Fumaric acid production with Rhizopus oryzae : quantifying time-dependent variations in catabolic flux
title_full_unstemmed Fumaric acid production with Rhizopus oryzae : quantifying time-dependent variations in catabolic flux
title_short Fumaric acid production with Rhizopus oryzae : quantifying time-dependent variations in catabolic flux
title_sort fumaric acid production with rhizopus oryzae quantifying time dependent variations in catabolic flux
topic UCTD
Rhizopus oryzae
Fumaric acid
Metabolic flux analysis
DO
PH
Immobilisation
Engineering, built environment and information technology theses SDG-09
Engineering, built environment and information technology theses SDG-12
Engineering, built environment and information technology theses SDG-17
url http://hdl.handle.net/2263/51118