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Effects of Temperature and pH on Recombinant Thaumatin II Production by Pichia pastoris

The sweet protein thaumatin is emerging as a promising sugar replacer in the market today, especially in the food and beverage sector. Rising demand for its production necessitates the large-scale extraction of this protein from its natural plant source, which can be limited in terms of raw material...

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Autores principales: Joseph, Jewel Ann, Akkermans, Simen, Van Impe, Jan F. M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9141780/
https://www.ncbi.nlm.nih.gov/pubmed/35627007
http://dx.doi.org/10.3390/foods11101438
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author Joseph, Jewel Ann
Akkermans, Simen
Van Impe, Jan F. M.
author_facet Joseph, Jewel Ann
Akkermans, Simen
Van Impe, Jan F. M.
author_sort Joseph, Jewel Ann
collection PubMed
description The sweet protein thaumatin is emerging as a promising sugar replacer in the market today, especially in the food and beverage sector. Rising demand for its production necessitates the large-scale extraction of this protein from its natural plant source, which can be limited in terms of raw material availability and production costs. Using a recombinant production technique via a yeast platform, specifically, Pichia pastoris, is more promising to achieve the product economically while maintaining batch-to-batch consistency. However, the bioproduction of recombinant proteins requires the identification of optimal process variables, constituting the maximal yield of the product of interest. These variables have a direct effect on the growth of the host organism and the secretion levels of the recombinant protein. In this study, two important environmental factors, pH, and temperature were assessed by cultivating P. pastoris in shake flasks to understand their influence on growth and the production levels of thaumatin II protein. The results from the pH study indicate that P. pastoris attained a higher viable cell density and secretion of protein at pH 6.0 compared to 5.0 when grown at 30 °C. Furthermore, within the three levels of temperatures investigated when grown at pH 6.0, the protein levels were the highest at 30 °C compared to 20 and 25 °C, whereas 25 °C exhibited the highest viable cell density. Interestingly, the trend observed from the qualitative effects of temperature and pH occurred in all the media that was investigated. These results broaden our understanding of how pH and temperature adjustment during P. pastoris cultivation aid in enhancing the production yields of thaumatin II prior to optimising the fed batch bioreactor operation.
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spelling pubmed-91417802022-05-28 Effects of Temperature and pH on Recombinant Thaumatin II Production by Pichia pastoris Joseph, Jewel Ann Akkermans, Simen Van Impe, Jan F. M. Foods Article The sweet protein thaumatin is emerging as a promising sugar replacer in the market today, especially in the food and beverage sector. Rising demand for its production necessitates the large-scale extraction of this protein from its natural plant source, which can be limited in terms of raw material availability and production costs. Using a recombinant production technique via a yeast platform, specifically, Pichia pastoris, is more promising to achieve the product economically while maintaining batch-to-batch consistency. However, the bioproduction of recombinant proteins requires the identification of optimal process variables, constituting the maximal yield of the product of interest. These variables have a direct effect on the growth of the host organism and the secretion levels of the recombinant protein. In this study, two important environmental factors, pH, and temperature were assessed by cultivating P. pastoris in shake flasks to understand their influence on growth and the production levels of thaumatin II protein. The results from the pH study indicate that P. pastoris attained a higher viable cell density and secretion of protein at pH 6.0 compared to 5.0 when grown at 30 °C. Furthermore, within the three levels of temperatures investigated when grown at pH 6.0, the protein levels were the highest at 30 °C compared to 20 and 25 °C, whereas 25 °C exhibited the highest viable cell density. Interestingly, the trend observed from the qualitative effects of temperature and pH occurred in all the media that was investigated. These results broaden our understanding of how pH and temperature adjustment during P. pastoris cultivation aid in enhancing the production yields of thaumatin II prior to optimising the fed batch bioreactor operation. MDPI 2022-05-16 /pmc/articles/PMC9141780/ /pubmed/35627007 http://dx.doi.org/10.3390/foods11101438 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Joseph, Jewel Ann
Akkermans, Simen
Van Impe, Jan F. M.
Effects of Temperature and pH on Recombinant Thaumatin II Production by Pichia pastoris
title Effects of Temperature and pH on Recombinant Thaumatin II Production by Pichia pastoris
title_full Effects of Temperature and pH on Recombinant Thaumatin II Production by Pichia pastoris
title_fullStr Effects of Temperature and pH on Recombinant Thaumatin II Production by Pichia pastoris
title_full_unstemmed Effects of Temperature and pH on Recombinant Thaumatin II Production by Pichia pastoris
title_short Effects of Temperature and pH on Recombinant Thaumatin II Production by Pichia pastoris
title_sort effects of temperature and ph on recombinant thaumatin ii production by pichia pastoris
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9141780/
https://www.ncbi.nlm.nih.gov/pubmed/35627007
http://dx.doi.org/10.3390/foods11101438
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