Introduction Folate or vitamin B9 is a water-soluble vitamin composed of several conjugated molecules namely pteridine ring, paraaminobenzoic acid (PABA), and glutamic acid. 1,2 Folate is required for normal cell fission and growth. It functions as a cofactor involved in various metabolic reactions in the body, such as synthesis, repair, DNA methylation reactions; nucleotide synthesis; and amino acid metabolism. 3,4 Folate is needed in a certain amount, especially during pregnancy and lactating period. The daily recommended intake of folate are 65 µg/d of dietary folate equivalents (DFEs) for 0-6 month infants as adequate intake (AI), 80 µg DFE for 7-12 month infants (AI), 150 µg DFE for 1-3 year old children as recommended dietary allowance (RDA), 200 µg DFE for 4-8 year old children (RDA), 300 µg DFE for 9-13 year old children (RDA), 400 µg DFE for 14-18 year old teens (RDA), 400 µg DFE for adults 19 years old and older (RDA), 500 µg DFE for lactating women (RDA), and 600 µg DFE for pregnant women (RDA). 5 If folate requirement is not sufficiently met, the body will experience folate deficiency and trigger various diseases such as anemia, neural tube defects, homocysteinemia, cardiovascular disease, and cancer. 4,6-9 Folate cannot be produced in the body, therefore, it must be obtained from food intake. It is naturally present in various types of food, such as cereals, fruits, vegetables, spices, nuts, eggs, and cheese. 10 However, natural folate has unstable properties, and its content is easily reduced during washing and processing. 11,12 Alternatively, folic acid, a synthetic form of folate, is generally chosen as the main source in fulfilling the daily needs as a food fortificant and food supplement due to its stability. 13,14 However, the metabolic process of folic acid in the body is relatively slow hence the body is not able to completely convert folic acid in large quantities. This results in a substantial accumulation of unmetabolized folic acid in cells. High levels of unmetabolized folic acid in the blood will cause a variety of metabolic disorders, such as masking symptoms of vitamin B12 deficiency, cognitive impairment, reducing the immune system, and cancer. 15,16 The emergence of health problems related to the use of synthetic folate has prompted many researchers to look for other, more stable, safer and more efficient sources of natural folate. Lactic acid bacteria (LAB) are known to produce folate both intracellularly and extracellularly. 12,17-19 Intracellular folate is in the intact cells, while extracellular folate is secreted to the growth medium. Extracellular folate is in the form of monoglutamate, thus it has a higher bioavailability compared to intracellular folate which is in the form of polyglutamate. Polyglutamate requires an enzymatic conjugation process