Carnosic Acid Production from Sugarcane Syrup by Engineered Yeast in Fed-Batch Fermentation

Phenolic diterpene carnosic acid (CA) is widely used in the food, nutritional health, and cosmetic industries due to its antioxidative and antimicrobial properties. This work aimed to overproduce CA in <i>Saccharomyces cerevisiae</i> from sugarcane syrup in fed-batch 2 L bioreactor ferme...

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Main Authors: Erdem Carsanba, Sara Fernandes, Felipe Beato, Luís Carlos Carvalho, Ana Pintado, Ana Lopes, Mónica Ribeiro, Tânia Leal, Manuela Pintado, Carla Oliveira
Format: Article
Language:English
Published: MDPI AG 2025-03-01
Series:Fermentation
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Online Access:https://www.mdpi.com/2311-5637/11/3/147
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author Erdem Carsanba
Sara Fernandes
Felipe Beato
Luís Carlos Carvalho
Ana Pintado
Ana Lopes
Mónica Ribeiro
Tânia Leal
Manuela Pintado
Carla Oliveira
author_facet Erdem Carsanba
Sara Fernandes
Felipe Beato
Luís Carlos Carvalho
Ana Pintado
Ana Lopes
Mónica Ribeiro
Tânia Leal
Manuela Pintado
Carla Oliveira
author_sort Erdem Carsanba
collection DOAJ
description Phenolic diterpene carnosic acid (CA) is widely used in the food, nutritional health, and cosmetic industries due to its antioxidative and antimicrobial properties. This work aimed to overproduce CA in <i>Saccharomyces cerevisiae</i> from sugarcane syrup in fed-batch 2 L bioreactor fermentation. A geranylgeranyl diphosphate (GGPP)-producing strain modified with genes encoding the enzymes copalyl diphosphate synthase (Pv.CPS), miltiradiene synthase (Ro.KSL2), hydroxy ferruginol synthase (Ro.HFS), CA synthase (Ro.CYP76AK8), CYP reductase (At.ATR1), and transketolase (TKL1) was used. Lowering the feed rate from 12–26 g/L/h to 7–8 g/L/h, and the use of a dynamic dissolved oxygen (DO) trigger (min. 10%, max. 40%, threshold 70%) instead of a DO trigger of 30%, enhanced CA production by 27%. As a result, the highest CA titer ever reported to date, 191.4 mg/L, was obtained in 4-day fermentation. This study shows the feasibility of engineered yeast to produce CA from the sustainable feedstock sugarcane syrup.
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series Fermentation
spelling doaj-art-d3c7297c1e5349628401fd2aa1dba3c42025-08-20T02:11:19ZengMDPI AGFermentation2311-56372025-03-0111314710.3390/fermentation11030147Carnosic Acid Production from Sugarcane Syrup by Engineered Yeast in Fed-Batch FermentationErdem Carsanba0Sara Fernandes1Felipe Beato2Luís Carlos Carvalho3Ana Pintado4Ana Lopes5Mónica Ribeiro6Tânia Leal7Manuela Pintado8Carla Oliveira9Amyris BioProducts Portugal, Unipessoal, Lda. Rua Diogo Botelho 1327, 4169-005 Porto, PortugalCBQF—Centro de Biotecnologia e Química Fina—Laboratório Associado, Escola Superior de Biotecnologia, Universidade Católica Portuguesa, Rua Diogo Botelho 1327, 4169-005 Porto, PortugalAmyris BioProducts Portugal, Unipessoal, Lda. Rua Diogo Botelho 1327, 4169-005 Porto, PortugalAmyris BioProducts Portugal, Unipessoal, Lda. Rua Diogo Botelho 1327, 4169-005 Porto, PortugalCBQF—Centro de Biotecnologia e Química Fina—Laboratório Associado, Escola Superior de Biotecnologia, Universidade Católica Portuguesa, Rua Diogo Botelho 1327, 4169-005 Porto, PortugalAmyris BioProducts Portugal, Unipessoal, Lda. Rua Diogo Botelho 1327, 4169-005 Porto, PortugalAmyris BioProducts Portugal, Unipessoal, Lda. Rua Diogo Botelho 1327, 4169-005 Porto, PortugalCBQF—Centro de Biotecnologia e Química Fina—Laboratório Associado, Escola Superior de Biotecnologia, Universidade Católica Portuguesa, Rua Diogo Botelho 1327, 4169-005 Porto, PortugalCBQF—Centro de Biotecnologia e Química Fina—Laboratório Associado, Escola Superior de Biotecnologia, Universidade Católica Portuguesa, Rua Diogo Botelho 1327, 4169-005 Porto, PortugalCBQF—Centro de Biotecnologia e Química Fina—Laboratório Associado, Escola Superior de Biotecnologia, Universidade Católica Portuguesa, Rua Diogo Botelho 1327, 4169-005 Porto, PortugalPhenolic diterpene carnosic acid (CA) is widely used in the food, nutritional health, and cosmetic industries due to its antioxidative and antimicrobial properties. This work aimed to overproduce CA in <i>Saccharomyces cerevisiae</i> from sugarcane syrup in fed-batch 2 L bioreactor fermentation. A geranylgeranyl diphosphate (GGPP)-producing strain modified with genes encoding the enzymes copalyl diphosphate synthase (Pv.CPS), miltiradiene synthase (Ro.KSL2), hydroxy ferruginol synthase (Ro.HFS), CA synthase (Ro.CYP76AK8), CYP reductase (At.ATR1), and transketolase (TKL1) was used. Lowering the feed rate from 12–26 g/L/h to 7–8 g/L/h, and the use of a dynamic dissolved oxygen (DO) trigger (min. 10%, max. 40%, threshold 70%) instead of a DO trigger of 30%, enhanced CA production by 27%. As a result, the highest CA titer ever reported to date, 191.4 mg/L, was obtained in 4-day fermentation. This study shows the feasibility of engineered yeast to produce CA from the sustainable feedstock sugarcane syrup.https://www.mdpi.com/2311-5637/11/3/147carnosic acidengineered <i>Saccharomyces cerevisiae</i>sugarcane syrupfed-batch fermentationdynamic dissolved oxygen trigger
spellingShingle Erdem Carsanba
Sara Fernandes
Felipe Beato
Luís Carlos Carvalho
Ana Pintado
Ana Lopes
Mónica Ribeiro
Tânia Leal
Manuela Pintado
Carla Oliveira
Carnosic Acid Production from Sugarcane Syrup by Engineered Yeast in Fed-Batch Fermentation
Fermentation
carnosic acid
engineered <i>Saccharomyces cerevisiae</i>
sugarcane syrup
fed-batch fermentation
dynamic dissolved oxygen trigger
title Carnosic Acid Production from Sugarcane Syrup by Engineered Yeast in Fed-Batch Fermentation
title_full Carnosic Acid Production from Sugarcane Syrup by Engineered Yeast in Fed-Batch Fermentation
title_fullStr Carnosic Acid Production from Sugarcane Syrup by Engineered Yeast in Fed-Batch Fermentation
title_full_unstemmed Carnosic Acid Production from Sugarcane Syrup by Engineered Yeast in Fed-Batch Fermentation
title_short Carnosic Acid Production from Sugarcane Syrup by Engineered Yeast in Fed-Batch Fermentation
title_sort carnosic acid production from sugarcane syrup by engineered yeast in fed batch fermentation
topic carnosic acid
engineered <i>Saccharomyces cerevisiae</i>
sugarcane syrup
fed-batch fermentation
dynamic dissolved oxygen trigger
url https://www.mdpi.com/2311-5637/11/3/147
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