We have investigated the impact of different yeasts and fermentation time on folate content and composition in a fermented maize-based porridge, called togwa, consumed in rural areas in Tanzania. The yeasts studied, originally isolated from indigenous togwa, belong to Issatchenkia orientalis, Pichia anomala, Saccharomyces cerevisiae, Klyveromyces marxianus and Candida glabrata. The main folate forms found, detected and quantified by HPLC during the fermentations were 5-methyl-tetrahydrofolate (5-CH(3)-H(4)folate) and tetrahydrofolate (H(4)folate). The content of H(4)folate, per unit togwa, remained fairly stable at a low level throughout the experiment for all strains, whereas the 5-CH(3)-H(4)folate concentration was highly dependent on yeast strain as well as on fermentation time. The highest folate concentration was found after 46 h of fermentation with C. glabrata (TY26) (6.91+/-0.14 microg 100 mL(-1)), corresponding to a 23-fold increase compared with unfermented togwa. The cell concentration per se could not predict the togwa folate level, as shown by the much higher specific folate content (g folate CFU(-1)) in the S. cerevisiae strain (TY08) compared with the other species tested. This study provides useful data when trying to maximize folate content in togwa as well as in other yeast-fermented products.

We here demonstrate that folate content in yeast fermented food can be dramatically increased by using a proper (i) yeast strain and (ii) cultivation procedure for the selected strain prior to food fermentation. Folate levels were 3 to 5-fold higher in white wheat bread leavened with a Saccharomyces cerevisiae strain CBS7764, cultured in defined medium and harvested in the respiro-fermentative phase of growth prior to dough preparation (135-139 mu g/100 dry matter), compared to white wheat bread leavened with commercial Baker's yeast (27-43 mu g/100 g). The commercial Baker's yeast strain had been industrially produced, using a fedbatch process. thereafter compressed and stored in the refrigerator until bakings were initiated. This strategy is an attractive alternative to fortification of bread with synthetically produced folic acid. By using a high folate producing strain cultured a suitable way folate levels obtained were in accordance with folic acid content in fortified cereal products. (C) 2008 Elsevier B.V. All rights reserved.

Folate content in a Saccharomyces cerevisiae strain was monitored during aerobic batch fermentation in synthetic growth medium, yeast peptone dextrose medium, and a molasses based medium. During growth in the synthetic medium large differences in intracellular folate content was observed at different phases. Specific folate levels, expressed per unit biomass, were highest during respiro-fermentative growth (120 mu g/g) and decreased during the respiratory and stationary phases. Thus, the physiological state of the cells clearly affects the folate content. This was confirmed in chemostat Cultures where total intracellular folate content increased linearly with increasing growth rate (r(2) = 0.998), indicating high growth rate i.e. respiro-fermentative growth to be most favourable to obtain high specific folate content. In complex media however, much lower folate content (15-40 mu g/g) was found throughout the batch growth. Only minor growth-phase related differences were detected. This shows the impact of cultivation medium on folate content in yeast. To further investigate which components that influence folate content, batch experiments in synthetic medium with addition of specific components were performed. Adding a raw mixture of peptides and amino acids (peptone) decreased folate levels extensively (90%) whereas adding amino acids one-by-one only had minor effects on the intracellular folate content. Furthermore, supplementing synthetic medium with pABA, folate or nucleotides did not change the intracellular folate content. This work constitutes the first steps towards an optimised process for production of natural folates for fortification Purposes, as well as an effort to gain fundamental understanding of folate requirements in yeast in relation to environmental conditions. (C) 2007 Elsevier B.V. All rights reserved.

For the first time to our knowledge, distribution and content of individual folate forms in kefir yeast strains were investigated. This was done using a validated method based on reversed-phase high performance liquid chromatography (HPLC) with fluorescence and diode array (DAD) detection. Eight kefir yeast strains, belonging to different Candida and Saccharomyces species, were isolated from Russian kefir granules. They were grown in synthetic media at standardized conditions before analysis. The average folate content for these yeast strains was 10,780±550 μg/100 g dry matter. In all yeast strains tested, the most abundant folate forms as percentages were 5-methyltetrahydrofolate (43–59%), and 5-formyltetrahydrofolate (23–38%), whereas tetrahydrafolate occurred in a lesser proportion (19–23%).

We have studied folate content and composition in 44 yeast strains cultivated in a synthetic medium at standardised conditions. Folates were measured by a validated HPLC method. The total folate content ranged from 4000 to 14,500 mu g/100g dry matter showing a large biodiversity among studied yeasts. The forms found were 5-CH3-H(4)folate and H(4)folate, also varying extensively in relative amounts between different strains. Several strains showed a showed two-fold or higher folate content as compared to the control strain-a commercial strain of Baker's yeast. This indicates that folate content in yeast-fermented foods may be more than doubled merely by choosing a proper strain.

A simplified HPLC method for rapid determination of folates in yeast with ultraviolet and fluorescence detection without sample purification has been developed. By use of the column Aquasil C-18, specially designed for polar analytes, and gradient elution, it was possible to separate and determine five folate derivatives: tetrahydrofolate, 5-methyltetrahydrofolate, and 5-formyltetrahydrofolate with fluorescence detection, and 10-formylfolic acid and folic acid with ultraviolet detection. The sample preparation required only a small amount of dry yeast (25-50 mg) and included an extraction of folates by heat treatment and deconjugation of folate polyglutamates to monoglutamates with the use of rat serum conjugase. Validation involved investigation of matrix effects, determination of recovery by standard addition method, repeatability, and stability tests. The dominating folate forms in commercial dry baker's yeast were found to be tetrahydrafolate and 5-methyltetrahydrofolate with a total folate content of 2890 mu g/100 g (63.4 nmol/g). The simplicity of the method makes it suitable for folate screening studies of different yeast strains.