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Engineered Bacillus subtilis for the de novo production of 2′-fucosyllactose

BACKGROUND: The most abundant human milk oligosaccharide in breast milk, 2′-fucosyllactose (2′-FL), has been approved as an additive to infant formula due to its multifarious nutraceutical and pharmaceutical functions in promoting neonate health. However, the low efficiency of de novo synthesis limi...

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Autores principales: Zhang, Quanwei, Liu, Zhenmin, Xia, Hongzhi, Huang, Ziyang, Zhu, Yonglian, Xu, Linfeng, Liu, Yanfeng, Li, Jianghua, Du, Guocheng, Lv, Xueqin, Liu, Long
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9164505/
https://www.ncbi.nlm.nih.gov/pubmed/35655274
http://dx.doi.org/10.1186/s12934-022-01838-w
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author Zhang, Quanwei
Liu, Zhenmin
Xia, Hongzhi
Huang, Ziyang
Zhu, Yonglian
Xu, Linfeng
Liu, Yanfeng
Li, Jianghua
Du, Guocheng
Lv, Xueqin
Liu, Long
author_facet Zhang, Quanwei
Liu, Zhenmin
Xia, Hongzhi
Huang, Ziyang
Zhu, Yonglian
Xu, Linfeng
Liu, Yanfeng
Li, Jianghua
Du, Guocheng
Lv, Xueqin
Liu, Long
author_sort Zhang, Quanwei
collection PubMed
description BACKGROUND: The most abundant human milk oligosaccharide in breast milk, 2′-fucosyllactose (2′-FL), has been approved as an additive to infant formula due to its multifarious nutraceutical and pharmaceutical functions in promoting neonate health. However, the low efficiency of de novo synthesis limits the cost-efficient bioproduction of 2′-FL. RESULTS: This study achieved 2′-FL de novo synthesis in a generally recognized as safe (GRAS) strain Bacillus subtilis. First, a de novo biosynthetic pathway for 2′-FL was introduced by expressing the manB, manC, gmd, wcaG, and futC genes from Escherichia coli and Helicobacter pylori in B. subtilis, resulting in 2′-FL production of 1.12 g/L. Subsequently, a 2′-FL titer of 2.57 g/L was obtained by reducing the competitive lactose consumption, increasing the regeneration of the cofactor guanosine-5′-triphosphate (GTP), and enhancing the supply of the precursor mannose-6-phosphate (M6P). By replacing the native promoter of endogenous manA gene (encoding M6P isomerase) with a constitutive promoter P7, the 2′-FL titer in shake flask reached 18.27 g/L. The finally engineered strain BS21 could produce 88.3 g/L 2′-FL with a yield of 0.61 g/g lactose in a 3-L bioreactor, without the addition of antibiotics and chemical inducers. CONCLUSIONS: The efficient de novo synthesis of 2′-FL can be achieved by the engineered B. subtilis, paving the way for the large-scale bioproduction of 2′-FL titer in the future. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12934-022-01838-w.
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spelling pubmed-91645052022-06-05 Engineered Bacillus subtilis for the de novo production of 2′-fucosyllactose Zhang, Quanwei Liu, Zhenmin Xia, Hongzhi Huang, Ziyang Zhu, Yonglian Xu, Linfeng Liu, Yanfeng Li, Jianghua Du, Guocheng Lv, Xueqin Liu, Long Microb Cell Fact Research BACKGROUND: The most abundant human milk oligosaccharide in breast milk, 2′-fucosyllactose (2′-FL), has been approved as an additive to infant formula due to its multifarious nutraceutical and pharmaceutical functions in promoting neonate health. However, the low efficiency of de novo synthesis limits the cost-efficient bioproduction of 2′-FL. RESULTS: This study achieved 2′-FL de novo synthesis in a generally recognized as safe (GRAS) strain Bacillus subtilis. First, a de novo biosynthetic pathway for 2′-FL was introduced by expressing the manB, manC, gmd, wcaG, and futC genes from Escherichia coli and Helicobacter pylori in B. subtilis, resulting in 2′-FL production of 1.12 g/L. Subsequently, a 2′-FL titer of 2.57 g/L was obtained by reducing the competitive lactose consumption, increasing the regeneration of the cofactor guanosine-5′-triphosphate (GTP), and enhancing the supply of the precursor mannose-6-phosphate (M6P). By replacing the native promoter of endogenous manA gene (encoding M6P isomerase) with a constitutive promoter P7, the 2′-FL titer in shake flask reached 18.27 g/L. The finally engineered strain BS21 could produce 88.3 g/L 2′-FL with a yield of 0.61 g/g lactose in a 3-L bioreactor, without the addition of antibiotics and chemical inducers. CONCLUSIONS: The efficient de novo synthesis of 2′-FL can be achieved by the engineered B. subtilis, paving the way for the large-scale bioproduction of 2′-FL titer in the future. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12934-022-01838-w. BioMed Central 2022-06-02 /pmc/articles/PMC9164505/ /pubmed/35655274 http://dx.doi.org/10.1186/s12934-022-01838-w Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Research
Zhang, Quanwei
Liu, Zhenmin
Xia, Hongzhi
Huang, Ziyang
Zhu, Yonglian
Xu, Linfeng
Liu, Yanfeng
Li, Jianghua
Du, Guocheng
Lv, Xueqin
Liu, Long
Engineered Bacillus subtilis for the de novo production of 2′-fucosyllactose
title Engineered Bacillus subtilis for the de novo production of 2′-fucosyllactose
title_full Engineered Bacillus subtilis for the de novo production of 2′-fucosyllactose
title_fullStr Engineered Bacillus subtilis for the de novo production of 2′-fucosyllactose
title_full_unstemmed Engineered Bacillus subtilis for the de novo production of 2′-fucosyllactose
title_short Engineered Bacillus subtilis for the de novo production of 2′-fucosyllactose
title_sort engineered bacillus subtilis for the de novo production of 2′-fucosyllactose
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9164505/
https://www.ncbi.nlm.nih.gov/pubmed/35655274
http://dx.doi.org/10.1186/s12934-022-01838-w
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