Short-term nitrogen limitation and continuous lighting (red/blue = 3:1) were applied individually and in combination to butterhead and red oak leaf lettuce for 1, 2, or 3 days before harvest to assess their effects on improving the nutritional value and sweet taste and reducing nitrate content and bitterness of lettuce. The results suggested that a 3-day nitrogen limitation combined with continuous lighting reduced the lettuce content of nitrate and sesquiterpene lactones and improved the quantities of soluble sugar, soluble protein, anthocyanins, and phenolic compounds without reducing the fresh weight of lettuce. In addition, in vitro simulated digestion results suggested that the 3-day nitrogen limitation combined with continuous lighting significantly improved the sweetness and reduced the bitterness of lettuce compared to the control. In conclusion, nitrogen limitation combined with continuous lighting for 3 days before harvest effectively enhanced the quality and taste of lettuce, showing great potential for its use in hydroponic lettuce production.