Background and ObjectivesThe production of high‐quality gluten‐free bread still poses a major technological challenge. The objective of this study was to investigate the effects of sourdough fermentation and structural/nutritional enrichment on gluten‐free sorghum bread quality. Lactic acid bacteria (Lactobacillus casei and Lactobacillus acidophilus) and yeast (Saccharomyces cerevisiae) were used as starter cultures.FindingsThe bread crust color and titratable acidity were comparable although the crumb color of sourdough bread made with L. acidophilus was measured darker. In both starter cultures, sourdough breads had a softer texture than the control, with significant (p<0.05) differences in hardness, adhesiveness, cohesiveness, and gumminess values. The sourdough breads showed similar FTIR spectra with the control bread, with four major peaks in the functional group region (4000‐1200 cm‐1) and a unique peak in the fingerprint region (1200‐600 cm‐1).ConclusionBreads made with L. acidophilus (at 20 and 30% addition levels) showed better quality attributes such as softer texture, higher total phenolic content (97.75 mg GAE/100 g), and antioxidant capacity (341.49 µmol TEAC/100 g).Significance and NoveltyThe results suggested that selected LAB‐based sorghum sourdough might be a promising ingredient for the development of gluten‐free bread with improved quality and nutritional value.This article is protected by copyright. All rights reserved.