Screening Of Folate Producing Bacteria From Injera Implication On Folate Production During Fermentation And Bioavailability.

Folate is an essential nutrient in the diets of humans involved in vital functions of cell metabolism. It isrncurrently one of the most actively studied vitamins. This is mainly due to its well established role inrnpreventing neural tube defects in the developing foetus. The present study investigated the potential ofrnusing a cereal-based fermented food (injera), made from tef to increase the folate intake of vulnerablernpopulation groups.rnTef flour (n=60), dough (n=60) and injera (n=60) were collected from the households of Addis (twornhousehold/sub-city). Total folate contents of tef flour, dough and injera was determined byrnmicrobiological assay using Lactobacillus rhamnosus (ATCC 7469) as the growth indicator organism torninvestigate the effect of processing on the total folate content. The contribution of tef injera consumptionrnto the recommended nutrient intake (RNI) of folate was estimated for children and women ofrnreproductive age. Numbering and isolation of potential folate producing LAB from fermented tef doughrnwere conducted to study their folate production capability in culture media. Identification of folaternproducing LAB was done morphologically by gram’s staining and genotypically using sequencing of 16SrnrRNA coding genes. The bioavailability of folate produced by the selected best folate producing LABrnstrain (Lactobacillus plantarum P2R3FA) was evaluated using a rat depletion-repletion assay. L.rnplantarum P2R3FA isolated from tef dough and commercial Saccharomyces cerevisiae were tested alonernor in combination for their ability to make injera with higher folate content. The folate productionrncapability of L. plantarum P2R3FA has been also evaluated for three successive batch fermentationsrnusing back-slopping. The acceptability of folate-enhanced injera was estimated by 30 adult healthyrnvolunteers, using a 9 point hedonic scale.rnThe average folate content (dry weight basis) of tef flour was 58.7 μg/100 g and injera made byrntraditional process using ersho per fresh weight was 14.3 μg/100 g, which can contribute up to arnmaximum of 10 % to RNI of folate for children (1-3 years) and women of reproductive age. The effect ofrnfermentation was variable as shown by folate retention, ranging from 60-148 %. Cooking led to folaternlosses (maximum of 52.8 %). The overall injera processing resulted in folate retention of 38-122 %.rnA total of 162 potential folate producing LAB were isolated, 19 of which were able to grow and producernfolate on folate-free culture medium. Total folate production of the isolates ranged from 1 to 43 μg/L.rnFour high folate producing isolates were selected, and based on 16S rRNA sequencing all of them werernmembers of the family Lactobacillaceae showing 98-100 % sequence identity to L. plantarum. The L.rnplantarum P2R3FA was able to restore normal blood folate status of folate-depleted rats.rnrnThe least folate content was observed in injera fermented by traditional process using ersho. Though thernfolate content of injera made with S. cerevisiae was the highest and contributed 23 and 27 % to folaternRNI of children and women of reproductive age respectively, it was the least accepted product by thernpotential consumers. Injera made with L. plantarum P2R3FA has also high amount of folate andrncontributed more than 15 % to the RNI. In addition, it was the most preferred product. The use of backslopingrnfrom the dough inoculated with L. plantarum P2R3FA was efficient since the folate content ofrninjera was still very high and the acceptance by the potential consumers was very good.

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