Folate, primarily derived from plants, plays a crucial role in plant growth and metabolism, and is closely associated with human health, including the prevention of birth defects, maintenance of neurological and fetal health, and treatment of anemia. Owing to its diverse biological activities, folate has been widely applied in the fields of medicine and food. Moringa (Moringa oleifera Lam.) is currently recognized as the plant with the highest known folate content and is extensively utilized in folate-related research. 5-Methyltetrahydrofolate (5-MTHF) is the only folate derivative that can directly participate in the methylation cycle, addressing the unmet needs of populations for whom synthetic folate is ineffective. However, 5-MTHF is present in plants at low levels and is challenging to extract. Consequently, with advancements in synthetic biology, novel approaches have emerged to provide greener, more efficient, and stable production pathways for 5-MTHF. This article systematically reviews the classification, biological activities, and biosynthetic pathways of folate, with a focus on biological strategies to enhance 5-MTHF production, aiming to deepen the understanding of its functional value. The review seeks to offer innovative perspectives for food science, nutrition, and related industries, while also exploring solutions for populations with special needs.

Isothiocyanates, secondary metabolites primarily derived from cruciferous plants, are known for their distinct flavors in food and diverse biological activities such as anti-tumor, anti-diabetic, neuroprotective, antibacterial, anti-inflammatory, and antioxidant properties. They are extensively utilized in food, medicine, and agriculture. Moringa oleifera Lam. has recently been recognized as a new food resource and has attracted increasing research attention. This plant belongs to the Brassicales order and the Moringa family. It was found to contain biologically active isothiocyanates such as sulforaphane and 4-(α-L-rhamnopyranosyloxy)-benzyl isothiocyanate (MIC-1). Considering the low content of natural products and the related environmental issues caused by chemical synthesis, synthetic biology presents a promising approach for the sustainable and efficient production of these compounds. Recently, the biosynthetic pathways of isothiocyanates in many plants have been elucidated, which can promote the heterologous production of isothiocyanates through microbial fermentation. This review comprehensively examines the bioactivity, bioavailability, and biosynthetic pathways of M. oleifera Lam. isothiocyanates, alongside recent advancements in molecular biology techniques aimed at enhancing their yield. By elucidating the functional significance of isothiocyanates, this review aims to provide a comprehensive understanding of their role and to support further developments in their utilization in functional foods. It also proposes strategies for achieving high-yield production of these valuable compounds, offering insights that bridge theoretical knowledge with practical applications in the field.