A method based on microbial re-inoculation, or the so-called backslopping and subsequent proofing of rye bread dough simulating commercial one-stage sourdough process, was used for the screening of the leavening capacity of sourdough yeast strains. Two yeast strains were initially tested with seven Lactobacillus strains. Thereafter, 17 yeast strains, mostly of sourdough origin, were tested with a backslopping procedure with heterofermentative Lactobacillus brevis as an acidifying lactic acid bacteria (LAB). The highest leavening capacity was found in sourdoughs containing Candida milleri, in particular when it was accompanied by obligately homofermentative Lactobacillus acidophilus or facultatively heterofermentative Lactobacillus plantarum when it acted homofermentatively. The leavening capacity of the reference strain Saccharomyces cerevisiae was about half that of C. milleri in all sourdoughs tested. The re-inoculation procedure increased the differences found in the leavening capacity of the tested yeast strains during final proofing of rye bread dough. The backslopped sourdoughs containing a heterofermentative Lactobacillus strain were more suppressive than those containing a homofermentative strain. The highest leavening capacity was found in C. milleri strains. The use of one backslopping cycle before assaying the leavening capacity of a laboratory sourdough is recommended since it helps to differentiate between yeast strains to be tested for their leavening power in the final bread dough.