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Engineered Probiotic Lactococcus lactis for Lycopene Production against ROS Stress in Intestinal Epithelial Cells.

  • 2022-03-15
  • ACS Synthetic Biology 11(4)
    • Jiapeng Wu
    • Xingfang Tian
    • Xiaoning Xu
    • X. Gu
    • J. Kong
    • T. Guo

Abstract

Lactococcus lactis is a food-grade chassis for delivery of bioactive molecules to the intestinal mucosa in situ, while its ability to produce lycopene for detoxification of reactive oxidative species (ROS) is not realized yet. Here, L. lactis NZ9000 was engineered to synthesize lycopene by heterologous expression of a gene cluster crtEBI in plasmids or chromosomes, yielding the recombinant strains NZ4 and NZ5 with 0.59 and 0.54 mg/L lycopene production, respectively. To reroute the pyruvate flux to lycopene, the main lactate dehydrogenase and α-acetolactate synthase pathways were sequentially disrupted. The resultant strains NZΔldh-1 and NZΔldhΔals-1 increased lycopene accumulation to 0.70 and 0.73 mg/L, respectively, while their biomasses were reduced by 12.42% and the intracellular NADH/NAD+ ratios increased by 3.05- and 2.10-fold. To increase the biomasses of these engineered strains, aerobic respiration was activated and tuned by the addition of exogenous heme and oxygen. As a result, the engineered L. lactis strains partly recovered the growth and redox balance, yielding the lycopene levels of 0.91-1.09 mg/L. The engineered L. lactis strain protected the intestinal epithelial cells NCM460 against H2O2 challenge, with a 30.09% increase of cell survival and a 29.2% decrease of the intracellular ROS level compared with strain NZ9000 treatment. In summary, this work established the use of the engineered probiotic L. lactis for lycopene production and prospected its potential in the prevention of intestinal oxidative damage.

Keywords: Lactococcus lactis; aerobic respiration; engineered probiotic; intestinal epithelial cells; lycopene; oxidative stress.

Research Insights

SupplementHealth OutcomeEffect TypeEffect Size
Lactococcus lactisImproved Cell Survival RatesBeneficial
Moderate
Lactococcus lactisReduced Intracellular Reactive Oxygen Species LevelsBeneficial
Moderate
Lactococcus lactis L1A-23Improved Cell Survival RatesBeneficial
Moderate
Lactococcus lactis L1A-23Reduced Intracellular Reactive Oxygen Species LevelsBeneficial
Moderate
Lactococcus lactis LL-23Reduced Oxidative Stress in Intestinal Epithelial CellsBeneficial
Moderate

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