Abstract Introduction Fermentation is an important biotechnological process that improves the biochemical properties of natural raw materials by transforming high molecular compounds into more beneficial low molecular compounds through microbial biotransformation. Ceylon cinnamon (Cinnamomum zeylanicum Blume), globally recognized as 'true cinnamon', contains high molecular weight, complex compounds that are responsible for its renowned antioxidant, antibacterial, and therapeutic properties, making it a vital component in medicinal applications1. Aims The present study aimed to investigate the effect of fermentation by Lactobacillus reuteri on selected phytochemical contents, antioxidant, and antibacterial properties of bark extracts of Ceylon cinnamon (Cinnamomum zeylanicum Blume) and to determine the optimum conditions for fermentation of Cinnamon bark extract by L. reuteri. Method Authenticated Ceylon cinnamon bark (CCB) was extracted into water using the decoction method. The water extract was fermented by L. reuteri under controlled conditions (pH 6, incubation at 35 ºC) and optimized using two parameters; the inoculum density (1.5×108 cfu/ml and 7.5×108 cfu/ml) and incubation time (2 and 4 days). After fermentation, total polyphenolic content (TPC), total flavonoid content (TFC), DPPH free radical scavenging activity and antibacterial activity of non-fermented extracts (NFE) and fermented extracts (FE) of CCB were evaluated. Antibacterial activity was determined using the agar well diffusion method against Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa. Ethical approval was not required since no human or animal study was done during the study. Results The extract fermented with an inoculum density of 7.5×108 cfu/ml and incubated for 4 days showed the highest TFC (19.3 ± 0.3 mg RE/g) and antioxidant activity (IC50: 97.2 ± 0.8 µg/ml) in comparison to that of NFEs (TFC:13.3 ± 0.3 mg RE/g and DPPH activity: IC50: 233.7 ± 12.8 µg/ml). Moreover, the fermented samples showed a reduction in TPC, indicating possible biotransformation of compounds. and no significant change in the antibacterial activity in comparison to the non-fermented samples. Discussion and Conclusion This study was mainly focused on investigating the effect of fermentation of Ceylon cinnamon bark aqueous extract by Lactobacillus reuteri. The increased antioxidant activity and total flavonoid content are notable and could be attributed to chemical transformations of some complex flavanols including catechin gallate and epicatechin gallate present in aqueous cinnamon to catechin and epicatechin monomers by the action of enzymes secreted by L. reuteri during the fermentation process2. Hence, this study suggests that fermentation using L. reuteri has a considerable positive impact on certain biochemical properties of cinnamon water extract. Cinnamon bark is a complex matrix containing various phytochemicals that could interact with bacterial strains in ways that are not fully understood. These interactions could lead to unexpected effects or limitations during fermentation.