Folate production in lactic acid bacteria and determination of folate derivatives by HPLC

Abstract

Folate is a B-group vitamin that humans cannot synthesize and must be taken externally. Although some types of Lactic Acid Bacteria can produce folate, there is insufficient information about the production of folate by yoghurt starter cultures. In this study, four bacteria belonging to two Lactobacillus, and two Streptococcus genera were isolated from yoghurt samples, and the isolates were biochemically identified by 16S rDNA with an API 50 CHL kit. Intracellular and extracellular folate production of the bacteria was designated in various media and buffer applications. The highest intracellular folate production was observed in Lactobacillus delbrueckii subsp. bulgaricus ZN951 (126.3 μg/L) and Streptococcus. thermophilus Z151 (177.1 μg/L) strains in MRS/Elliker medium with potassium phosphate buffer. L. delbrueckii subsp. bulgaricus ZN541 strain showed the highest extracellular folate production (105.1μg/L) in the same medium and buffer, but S. thermophilus Z151 strain exhibited 86.4 μg/L folate productions. Whether the strains contained folate derivatives and pABA was investigated by HPLC analysis and pABA was not detected in any of the strains. Among cultures, the highest total folate content was detected as 110.5μg/L by L. delbrueckii subsp. bulgaricus ZN951 strain. This strain was found to have concentrations of 5-MeTHF and 5-FmTHF. Therefore, the use of these bacteria can be an alternative to naturally increase folate content in foods.

Keywords

• Intracellular folate
• Extracellular folate
• Folate derivates
• Lactic acid bacteria

Laktik asit bakterilerinde folat üretimi ve HPLC ile folat türevlerinin tayini

Öz

Folat, insanların sentezleyemedikleri ve dışarıdan alınması gereken B grubu bir vitamindir. Bazı Laktik Asit Bakterileri folat üretebilse de, yoğurt starter kültürlerinin folat üretimi hakkında yeterli bilgi yoktur. Bu çalışmada yoğurt örneklerinden iki Lactobacillus ve iki Streptococcus cinsine ait dört bakteri izole edilmiş ve izolatlar API 50 CHL kiti ile 16S rDNA ile biyokimyasal olarak tanımlanmıştır. Bakterilerin hücre içi ve hücre dışı folat üretimi çeşitli ortam ve tampon uygulamalarında belirlenmiştir. En yüksek hücre içi folat üretimi Lactobacillus delbrueckii subsp. bulgaricus ZN951 (126.3 μg/L) ve Streptococcus thermophilus Z151 (177.1 μg/L) suşlarında MRS/Elliker besiortamı ve potasyum fosfat tamponunda belirlenmiştir. L. delbrueckii subsp. bulgaricus ZN541 suşu, aynı ortam ve tamponda en yüksek hücre dışı folat üretimini (105.1 μg/L) gösterirken, S. thermophilus Z151 suşu, 86.4 μg/L folat üretimi sergilemiştir. Suşların folat türevleri ve pABA içerip içermediği HPLC analizi ile araştırılmış ve suşların hiçbirinde pABA tespit edilememiştir. Kültürler arasında en yüksek toplam folat içeriği 110.5 μg/L olarak L. delbrueckii subsp. bulgaricus ZN951 suşunda tespit edilmiştir. Bu suşun 5-MeTHF ve5-FmTHF konsantrasyonlarına sahip olduğu bulunmuştur. Bu nedenle, bu bakterilerin kullanımı gıdalardaki folat içeriğini doğal olarak arttırmaya bir alternatif olabilir.

Anahtar Kelimeler

• ücre içi folat
• Hücre dişi folat
• Folat türevleri
• Laktik asit bakterisi

Kaynakça

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