Grain lysine enrichment and improved stress tolerance in rice through protein engineering

Amino acids are a major source of nourishment for people living in regions where rice is a staple food. However, rice grain is deficient in essential amino acids including lysine. The activity of the enzyme dihydrodipicolinate synthase (DHDPS) is crucial for lysine production in higher plants, but it is tightly regulated through feedback inhibition by its end product, lysine, leading to limited activity in the grain and resulting in low lysine accumulation. We identified lysine binding sites in the DHDPS enzyme and introduced key mutations to make DHDPS lysine feedback insensitive. Using in vivo analysis and functional complementation assays, we confirmed that protein engineering of the DHDPS renders it insensitive to lysine. Expression of mutated DHDPS resulted in 29% higher lysine and 15% higher protein accumulation in rice grains than in the wild type. Importantly, the lysine content in transgenic grains was maintained in cooked rice. The transgenic plants also exhibited enhanced stress tolerance along with higher antioxidant levels, improved photosynthesis, and higher grain yield compared to wild-type plants. We have shown that protein engineering of DHDPS in rice can lead to accumulation of lysine in grains and impart abiotic stress tolerance. This approach could improve health in regions with nutrient deficiencies and environmental stressors that challenge food production and human health.