The heavy use of mineral fertilizers causes imbalances in the biological processes that take place in soil. Therefore, it is necessary to develop more effective fertilizers or fertilizer complexes that ensure agricultural productivity and soil conservation. There is currently a lack of knowledge regarding the effectiveness of biologically enriched, complex mineral fertilizers for spring barley fertilization. The hypothesis of this study was that bacteria-enriched (Paenibacillus azotofixans, Bacillus megaterium, Bacillus mucilaginosus, and Bacillus mycoides), complex mineral fertilizers (N5P20.5K36) have significant impacts on the yield and potential for economic use of spring barley. Experimental studies were carried out for three years (2020–2022) with sandy loam soil in southern Lithuania. Four different spring barley fertilization scenarios (SCs) were investigated. In SC-1 (control), complex mineral fertilizer (N5P20.5K36) was not applied. In the other SCs, spring barley was sown with a drill and fertilizers were incorporated locally during the sowing operation: fertilization scenario SC-2 used 300 kg ha−1, SC-3 used 150 kg ha−1 preceded by a bacteria-inoculated complex mineral fertilizer (N5P20.5K36), and SC-4 used 300 kg ha−1 with the same bacterial complex. The results showed that the bacterial inoculant increased the efficiency of the mineral fertilizer and had an effect on plant growth in barley. For three consecutive years in the same plots, the bacterial inoculant showed significant positive effects on grain yield (changes of 8.1% in 2020, 6.8% in 2021, and 17.3% in 2022 between SC-2 and SC-4). Comparing the several different fertilizer scenarios from an economic point of view, it was observed that the highest profit per hectare was obtained with SC-4 in all three years of the study. Comparing SC-4 and SC-2, an increase of 13.7% was observed in 2020, followed by 9.1% and 41.9% in 2021 and in 2022, respectively. This study will be useful for farmers, biological inoculant manufacturers, and scientists researching the effectiveness of biological inoculants for growing agricultural crops. We found that it is possible to increase the yield of barley (7–17%) using the same rate of mineral fertilization by enriching it with bacterial inoculants. Further studies should be conducted to determine the effects of the bacterial inoculant on crop yield and soil over a period longer than 3 years.