β-Galactosidase expression on Lactococcus sp. KTH0-1S as a whole-cell biocatalyst for efficient lactosucrose production
- 1Department of Biotechnology, Faculty of Agro-Industry, Kasetsart University, Bangkok, Thailand
- 2Biocatalyst Technology Research Team, National Center of Genetic Engineering and Biotechnology (BIOTEC), Pathum Thani, Thailand
- 3Food Biotechnology Laboratory, Department of Biotechnology and Food Sciences, BOKU University, Vienna, Austria
Abstract
Lactosucrose is a functional trisaccharide with prebiotic properties and growing potential in food and agricultural applications; however, its industrial production is limited by low conversion efficiency and high downstream processing costs. In this study, a whole-cell biocatalyst was developed by displaying β-galactosidase on Lactococcus sp. KTH0-1S using a nisin-inducible expression system, the USP45 signal peptide, and a LysM anchoring domain. The engineered strain exhibited a maximum whole-cell β-galactosidase activity of 146.0 Miller units after 4 h of induction. The whole-cell enzyme efficiently catalyzed lactosucrose synthesis from lactose and sucrose, with optimal activity observed at pH 6.0 and 50 °C. Substrate molar ratio significantly influenced product formation, and the highest lactosucrose concentration (15,940 mg/L) was achieved at a lactose:sucrose ratio of 1:3. Reusability studies revealed high initial catalytic activity but limited stability upon repeated reaction cycles, likely due to the non-covalent nature of LysM-mediated surface anchoring. Molecular docking analysis supported the experimental observations by identifying a high-affinity substrate-binding pocket capable of accommodating both donor and acceptor substrates, providing a structural rationale for enhanced transgalactosylation under sucrose-enriched conditions. Overall, this study demonstrates the feasibility of using a whole-cell biocatalyst based on Lactococcus sp. for efficient lactosucrose production and highlights its potential as a cost-effective and food-compatible platform for the synthesis of functional oligosaccharides.
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BibTeX
@article{ariyakajorn2026βgalactosidase,
title = {{β-Galactosidase expression on Lactococcus sp. KTH0-1S as a whole-cell biocatalyst for efficient lactosucrose production}},
author = {Nichapat Ariyakajorn and Napat Sunsern and Phitsanu Pinmanee and Boontiwa Ninchan and Pimsiriya Srila and Konlarat Phirom-on and Sivatch Imrattanarak and Wirat Vanichsriratana and Nisit Watthanasakphuban},
journal = {PLOS One},
year = {2026},
month = sep,
doi = {10.1371/journal.pone.0359091},
url = {https://doi.org/10.1371/journal.pone.0359091},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - β-Galactosidase expression on Lactococcus sp. KTH0-1S as a whole-cell biocatalyst for efficient lactosucrose production
AU - Nichapat Ariyakajorn
AU - Napat Sunsern
AU - Phitsanu Pinmanee
AU - Boontiwa Ninchan
AU - Pimsiriya Srila
AU - Konlarat Phirom-on
AU - Sivatch Imrattanarak
AU - Wirat Vanichsriratana
AU - Nisit Watthanasakphuban
PY - 2026
DA - 2026/09/25
JO - PLOS One
DO - 10.1371/journal.pone.0359091
UR - https://doi.org/10.1371/journal.pone.0359091
AB - Lactosucrose is a functional trisaccharide with prebiotic properties and growing potential in food and agricultural applications; however, its industrial production is limited by low conversion efficiency and high downstream processing costs. In this study, a whole-cell biocatalyst was developed by displaying β-galactosidase on Lactococcus sp. KTH0-1S using a nisin-inducible expression system, the USP45 signal peptide, and a LysM anchoring domain. The engineered strain exhibited a maximum whole-cell β-galactosidase activity of 146.0 Miller units after 4 h of induction. The whole-cell enzyme efficiently catalyzed lactosucrose synthesis from lactose and sucrose, with optimal activity observed at pH 6.0 and 50 °C. Substrate molar ratio significantly influenced product formation, and the highest lactosucrose concentration (15,940 mg/L) was achieved at a lactose:sucrose ratio of 1:3. Reusability studies revealed high initial catalytic activity but limited stability upon repeated reaction cycles, likely due to the non-covalent nature of LysM-mediated surface anchoring. Molecular docking analysis supported the experimental observations by identifying a high-affinity substrate-binding pocket capable of accommodating both donor and acceptor substrates, providing a structural rationale for enhanced transgalactosylation under sucrose-enriched conditions. Overall, this study demonstrates the feasibility of using a whole-cell biocatalyst based on Lactococcus sp. for efficient lactosucrose production and highlights its potential as a cost-effective and food-compatible platform for the synthesis of functional oligosaccharides.
SN - 1932-6203
ER -