Efficient metabolic pathway modification in various strains of lactic acid bacteria using CRISPR/Cas9 system for elevated synthesis of antimicrobial compounds

Lactic acid bacteria (LAB) are known to exhibit various beneficial roles in fermentation, serving as probiotics, and producing a plethora of valuable compounds including antimicrobial activity such as bacteriocin-like inhibitory substance (BLIS) that can be used as biopreservative to improve food sa...

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Main Authors: Haryani, Yuli, Abdul Halid, Nadrah, Goh, Sur Guat, Mahmud Ab Rashid, Nor-Khaizura, Md Hatta, Muhammad Asyraf, Sabri, Suriana, Radu, Son, Hasan, Hanan
Format: Article
Language:English
Published: Elsevier 2024
Online Access:http://psasir.upm.edu.my/id/eprint/114353/
http://psasir.upm.edu.my/id/eprint/114353/1/114353.pdf
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author Haryani, Yuli
Abdul Halid, Nadrah
Goh, Sur Guat
Mahmud Ab Rashid, Nor-Khaizura
Md Hatta, Muhammad Asyraf
Sabri, Suriana
Radu, Son
Hasan, Hanan
author_facet Haryani, Yuli
Abdul Halid, Nadrah
Goh, Sur Guat
Mahmud Ab Rashid, Nor-Khaizura
Md Hatta, Muhammad Asyraf
Sabri, Suriana
Radu, Son
Hasan, Hanan
author_sort Haryani, Yuli
building UPM Institutional Repository
collection Online Access
description Lactic acid bacteria (LAB) are known to exhibit various beneficial roles in fermentation, serving as probiotics, and producing a plethora of valuable compounds including antimicrobial activity such as bacteriocin-like inhibitory substance (BLIS) that can be used as biopreservative to improve food safety and quality. However, the yield of BLIS is often limited, which poses a challenge to be commercially competitive with the current preservation practice. Therefore, the present work aimed to establish an optimised two-plasmid CRISPR/Cas9 system to redirect the carbon flux away from lactate towards compounds with antimicrobial activity by disrupting lactate dehydrogenase gene (ldh) on various strains of LAB. The lactic acid-deficient (ldhΔ) strains caused a metabolic shift resulting in increased inhibitory activity against selected foodborne pathogens up to 78 % than the wild-type (WT) strain. The most significant effect was depicted by Enterococcus faecalis-ldh∆ which displayed prominent bactericidal effects against all foodborne pathogens as compared to the WT that showed no antimicrobial activity. The present work provided a framework model for economically important LAB and other beneficial bacteria to synthesise and increase the yield of valuable food and industrial compounds. The present work reported for the first time that the metabolism of selected LAB can be manipulated by modifying ldh to attain metabolites with higher antimicrobial activity.
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institution Universiti Putra Malaysia
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language English
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spelling upm-1143532025-03-10T01:27:27Z http://psasir.upm.edu.my/id/eprint/114353/ Efficient metabolic pathway modification in various strains of lactic acid bacteria using CRISPR/Cas9 system for elevated synthesis of antimicrobial compounds Haryani, Yuli Abdul Halid, Nadrah Goh, Sur Guat Mahmud Ab Rashid, Nor-Khaizura Md Hatta, Muhammad Asyraf Sabri, Suriana Radu, Son Hasan, Hanan Lactic acid bacteria (LAB) are known to exhibit various beneficial roles in fermentation, serving as probiotics, and producing a plethora of valuable compounds including antimicrobial activity such as bacteriocin-like inhibitory substance (BLIS) that can be used as biopreservative to improve food safety and quality. However, the yield of BLIS is often limited, which poses a challenge to be commercially competitive with the current preservation practice. Therefore, the present work aimed to establish an optimised two-plasmid CRISPR/Cas9 system to redirect the carbon flux away from lactate towards compounds with antimicrobial activity by disrupting lactate dehydrogenase gene (ldh) on various strains of LAB. The lactic acid-deficient (ldhΔ) strains caused a metabolic shift resulting in increased inhibitory activity against selected foodborne pathogens up to 78 % than the wild-type (WT) strain. The most significant effect was depicted by Enterococcus faecalis-ldh∆ which displayed prominent bactericidal effects against all foodborne pathogens as compared to the WT that showed no antimicrobial activity. The present work provided a framework model for economically important LAB and other beneficial bacteria to synthesise and increase the yield of valuable food and industrial compounds. The present work reported for the first time that the metabolism of selected LAB can be manipulated by modifying ldh to attain metabolites with higher antimicrobial activity. Elsevier 2024 Article PeerReviewed text en http://psasir.upm.edu.my/id/eprint/114353/1/114353.pdf Haryani, Yuli and Abdul Halid, Nadrah and Goh, Sur Guat and Mahmud Ab Rashid, Nor-Khaizura and Md Hatta, Muhammad Asyraf and Sabri, Suriana and Radu, Son and Hasan, Hanan (2024) Efficient metabolic pathway modification in various strains of lactic acid bacteria using CRISPR/Cas9 system for elevated synthesis of antimicrobial compounds. Journal of Biotechnology, 395. pp. 53-63. ISSN 0168-1656; eISSN: 1873-4863 https://linkinghub.elsevier.com/retrieve/pii/S0168165624002396 10.1016/j.jbiotec.2024.09.002
spellingShingle Haryani, Yuli
Abdul Halid, Nadrah
Goh, Sur Guat
Mahmud Ab Rashid, Nor-Khaizura
Md Hatta, Muhammad Asyraf
Sabri, Suriana
Radu, Son
Hasan, Hanan
Efficient metabolic pathway modification in various strains of lactic acid bacteria using CRISPR/Cas9 system for elevated synthesis of antimicrobial compounds
title Efficient metabolic pathway modification in various strains of lactic acid bacteria using CRISPR/Cas9 system for elevated synthesis of antimicrobial compounds
title_full Efficient metabolic pathway modification in various strains of lactic acid bacteria using CRISPR/Cas9 system for elevated synthesis of antimicrobial compounds
title_fullStr Efficient metabolic pathway modification in various strains of lactic acid bacteria using CRISPR/Cas9 system for elevated synthesis of antimicrobial compounds
title_full_unstemmed Efficient metabolic pathway modification in various strains of lactic acid bacteria using CRISPR/Cas9 system for elevated synthesis of antimicrobial compounds
title_short Efficient metabolic pathway modification in various strains of lactic acid bacteria using CRISPR/Cas9 system for elevated synthesis of antimicrobial compounds
title_sort efficient metabolic pathway modification in various strains of lactic acid bacteria using crispr/cas9 system for elevated synthesis of antimicrobial compounds
url http://psasir.upm.edu.my/id/eprint/114353/
http://psasir.upm.edu.my/id/eprint/114353/
http://psasir.upm.edu.my/id/eprint/114353/
http://psasir.upm.edu.my/id/eprint/114353/1/114353.pdf