Background and objectivesExtensive research studies worldwide have discussed and analyzed the sourdough nutritional, chemical, and biotechnological aspects of western types of bread. However, the literature on sourdough usage in Arabic bread processing is very limited. The present study aimed to come up with a new fermentation technique at the bakery level. The new technique is hoped to reveal the traditional nostalgic characteristic flavor of Arabic bread produced using natural sourdough instead of the traditional yeasted dough and study its effect on the resulting bread. Three types of flour (straight grade, whole wheat, and rye) that are ordinarily used in Jordan were used to prepare three natural sour starters using the backslopping technique. FindingsDominant yeast species isolated from the traditional sour starter of the local Jordanian whole wheat were Saccharomyces cerevisiae, Candida glabrata, and Zygosaccharomyces spp., while rye and straight-grade starters showed different domination patterns. The dominant species of lactic acid bacteria (LAB) isolated from the traditional sour starter of the local Jordanian whole wheat (Horani cultivar) were Lactiplantibacillus plantarum2, Lactobacillus delbrueckii ssp bulgaricus, and the Lacticaseibacillus paracasei ssp paracasei3, while mixed strains of LAB isolates were obtained for the rye and straight-grade flour starters on day 7 of preparation.The pH results showed no significant differences (P > 0.05) between the three types of sour starters, while the total titratable acidity (TTA) results showed a significant increase (P ≤ 0.05) in whole wheat sour starters with a TTA value of 11.17 ± 0.11 ml 0.1 N NaOH/10g dough. The 20% and 30% straight-grade sour bread samples and the control exhibited the highest scores in terms of acceptability and total quality scores. ConclusionsThe incorporation of sourdough in Arabic flatbread processing showed remarkable results in extending the shelf life by using natural ingredients, improving the quality, and providing tastier and more convenient bread.