Cocoa Pulp Juice as a Fermentation Substrate for Potential Probiotic Lacticaseibacillus paracasei WU2502: Genomic Characterization, Metabolite Profiling, and Antimicrobial Activity Against Foodborne Pathogens
8 affiliations
- 1Akkhraratchakumari Veterinary College, Walailak University, Nakhon Si Thammarat 80160, Thailand
- 2Research Center in One Health, Walailak University, Nakhon Si Thammarat 80160, Thailand
- 3Research Center of Excellence in Innovation of Essential Oil Center of Excellence in Innovation of Essential Oil and Bio-Active Compound, Walailak University, Nakhon Si Thammarat 80160, Thailand
- 4Department of Family and Preventive Medicine, Faculty of Medicine, Prince of Songkla University, Hat Yai, Songkhla 90110, Thailand
- 5Department of Microbiology, Faculty of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran
- 6Department of Medical Technology, School of Allied Health Sciences, Walailak University, Nakhon Si Thammarat 80160, Thailand
- 7Research Center for Microbiome, Systems Biology, and Medical Innovation, Walailak University, Nakhon Si Thammarat 80160, Thailand
- 8Research Center in Tropical Pathobiology, Walailak University, Nakhon Si Thammarat 80160, Thailand
Abstract
Cocoa pulp juice is an underutilized by-product generated during cocoa bean fermentation that is rich in fermentable sugars and phytochemicals but remains largely unexplored as a substrate for microbial fermentation. This study aimed to evaluate cocoa pulp juice as a sustainable fermentation substrate for the potential probiotic Lacticaseibacillus paracasei WU2502 using whole-genome sequencing (WGS), carbohydrate-active enzyme (CAZyme) analysis, untargeted LC–QTOF-MS-based metabolite profiling, and antimicrobial evaluation. L. paracasei WU2502 showed substantial growth in cocoa pulp juice, reaching a maximum viable count of 10.70 log CFU/mL in 25% juice and >11 log CFU/mL in 50% and 100% juice, while the pH decreased from approximately 3.9–4.0 at Day 0 to 3.1–3.2 during fermentation. Fermented cocoa pulp juice showed antibacterial activity against Bacillus cereus, Staphylococcus aureus, Salmonella enterica serovar Typhimurium and Escherichia coli O157:H7 with MIC and MBC values of 12.5–25% (v/v). In a preliminary qualitative assessment, neutralization reduced the observed inhibitory activity, whereas Proteinase K treatment produced little apparent change, suggesting a possible pH-dependent contribution to the antimicrobial activity. WGS resulted in a 3.02 Mb draft genome with a GC content of 46.27%, 3114 coding sequences and genes relevant to glucose metabolism, acid tolerance, oxidative stress response and antimicrobial activity. Genome mining identified putative bacteriocin-associated regions containing Carnocin CP52- and Thermophilin A-related sequences and a RiPP-like biosynthetic region. No acquired antimicrobial resistance genes or virulence-associated factors were detected. CAZyme analysis revealed glycoside hydrolases and glycosyltransferases that may be involved in carbohydrate metabolic pathways and adaptation to plant-derived substrates. LC–MS/MS analysis of unfermented cocoa pulp juice provided baseline chemical characterization of the starting substrate, with representative putative annotations including sucrose, maltotriose, myo-inositol, citric acid, pyroglutamic acid, and choline. These findings support the feasibility of cocoa pulp juice as a fermentation substrate for L. paracasei WU2502 and provide a basis for further evaluation of this strain and fermented matrix for potential food applications.
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BibTeX
@article{mitsuwan2026cocoa,
title = {{Cocoa Pulp Juice as a Fermentation Substrate for Potential Probiotic Lacticaseibacillus paracasei WU2502: Genomic Characterization, Metabolite Profiling, and Antimicrobial Activity Against Foodborne Pathogens}},
author = {Watcharapong Mitsuwan and Phoomjai Sornsenee and Morteza Saki and Chonticha Romyasamit},
journal = {International Journal of Molecular Sciences},
year = {2026},
month = oct,
doi = {10.3390/ijms27219795},
url = {https://www.mdpi.com/1422-0067/27/21/9795},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - Cocoa Pulp Juice as a Fermentation Substrate for Potential Probiotic Lacticaseibacillus paracasei WU2502: Genomic Characterization, Metabolite Profiling, and Antimicrobial Activity Against Foodborne Pathogens
AU - Watcharapong Mitsuwan
AU - Phoomjai Sornsenee
AU - Morteza Saki
AU - Chonticha Romyasamit
PY - 2026
DA - 2026/10/31
JO - International Journal of Molecular Sciences
DO - 10.3390/ijms27219795
UR - https://www.mdpi.com/1422-0067/27/21/9795
AB - Cocoa pulp juice is an underutilized by-product generated during cocoa bean fermentation that is rich in fermentable sugars and phytochemicals but remains largely unexplored as a substrate for microbial fermentation. This study aimed to evaluate cocoa pulp juice as a sustainable fermentation substrate for the potential probiotic Lacticaseibacillus paracasei WU2502 using whole-genome sequencing (WGS), carbohydrate-active enzyme (CAZyme) analysis, untargeted LC–QTOF-MS-based metabolite profiling, and antimicrobial evaluation. L. paracasei WU2502 showed substantial growth in cocoa pulp juice, reaching a maximum viable count of 10.70 log CFU/mL in 25% juice and >11 log CFU/mL in 50% and 100% juice, while the pH decreased from approximately 3.9–4.0 at Day 0 to 3.1–3.2 during fermentation. Fermented cocoa pulp juice showed antibacterial activity against Bacillus cereus, Staphylococcus aureus, Salmonella enterica serovar Typhimurium and Escherichia coli O157:H7 with MIC and MBC values of 12.5–25% (v/v). In a preliminary qualitative assessment, neutralization reduced the observed inhibitory activity, whereas Proteinase K treatment produced little apparent change, suggesting a possible pH-dependent contribution to the antimicrobial activity. WGS resulted in a 3.02 Mb draft genome with a GC content of 46.27%, 3114 coding sequences and genes relevant to glucose metabolism, acid tolerance, oxidative stress response and antimicrobial activity. Genome mining identified putative bacteriocin-associated regions containing Carnocin CP52- and Thermophilin A-related sequences and a RiPP-like biosynthetic region. No acquired antimicrobial resistance genes or virulence-associated factors were detected. CAZyme analysis revealed glycoside hydrolases and glycosyltransferases that may be involved in carbohydrate metabolic pathways and adaptation to plant-derived substrates. LC–MS/MS analysis of unfermented cocoa pulp juice provided baseline chemical characterization of the starting substrate, with representative putative annotations including sucrose, maltotriose, myo-inositol, citric acid, pyroglutamic acid, and choline. These findings support the feasibility of cocoa pulp juice as a fermentation substrate for L. paracasei WU2502 and provide a basis for further evaluation of this strain and fermented matrix for potential food applications.
SN - 1422-0067
ER -