Development of therapeutic or prophylactic recombinant Lactococcus lactis NZ9000-fermented milk with KRAS mimotope

ABSTRACT

Milk is a common carrier for live cultures due to its abundant nutrients. Accumulating evidence has shown the efficacy of live culture as a vector to deliver drugs. This study aimed to develop recombinant Lactococcus lactis NZ9000-fermented milk expressing KRAS-139A-TTD mimotope. The fermentation conditions were optimized with inoculum concentration (6.4, 7, 8 log₁₀ CFU/mL), temperature (22–25, 30, 37°C), and time (3, 16, 24, 48 h). The optimized L. lactis-fermented milk (8 log₁₀ CFU/mL inoculum, 48 h, and 30°C) had a viability of 9.79 log₁₀ CFU/mL, pH 5.16, and 0.54% titratable acidity. There was an absence of post-acidification and syneresis, with a 17.3% viability reduction after 26 days of storage at 4°C. Furthermore, the KRAS peptides expression in fermented milk after 10 days of storage was detected through SDS-PAGE. This showed that L. lactis NZ9000-fermented milk has the potential as KRAS positive therapeutic or prophylactic functional drink.

KEYWORDS:

• Functional fermented milk
• KRAS peptide
• mimotope
• NICE system
• oral vaccine
• recombinant Lactococcus lactis

Acknowledgments

The authors would like to express their gratitude towards Professor Dr. Raha Abdul Rahim from University Putra Malaysia for providing the Lactococcus lactis NZ9000 used in this study.

Disclosure statement

No potential conflict of interest was reported by the author(s).

Data availability

The data that support the findings of this study are available from the corresponding authors upon reasonable request.

Author’s contribution

Shi-Yie Tan: Data curation, Investigation, Writing-original draft; Yu-Hsuan How: Writing-review & editing, Visualisation; Pui-Yan Siak: Writing-review & editing; Liew-Phing Pui: Supervision, Funding acquisition, Project administration; Lionel Lian-Aun In: Supervision, Funding acquisition, Conceptualization

Additional information

Funding

This work was supported by the UCSI University through UCSI Research Excellence & Innovative Grant (REIG) under REIG-FAS-2020/003,REIG-FPS-2020/060.