The effects of red beetroot juice fermentation by Lactobacillus acidophilus LA-5 on the activity of angiotensin-converting enzyme (ACE), acetylcholinesterase (AChE), and antioxidants, and its relationship with active substances were studied. The number of viable bacteria at the end of fermentation was 6.29×108 CFU/mL. The contents of betacyanins and total acid increased by 13.63% and 302.21%, respectively. The contents of reducing sugar, betaxanthins, and flavones decreased by 63.87%, 43.09%, and 0.05%, respectively. After fermentation, the clearance rates of ·O2-, ·OH, and ABTS+· increased by 29.13%, 96.36%, and 77.49%, respectively, and the inhibition rates of AChE and ACE increased by 59.15% and 35.37%, respectively. Twelve betalains were identified by UHPLC-Q-TOF-MS/MS. Correlation analysis showed that the aforementioned functional indexes had a very significant positive correlation with the betacyanin content (p<0.01). Three principal components, with a total contribution rate of 90.63%, were extracted using principal component analysis. The main active components were betanin, betanidin, 17-decarboxy-betanidin, 2-decarboxy-neobotanin, 6'-O-ferroyl-betanin, vulgaxanthin I, and betalamic acid. Q-type cluster analysis classified the fermentation process into three categories: slow growth, significant change, and flat periods. R-type cluster analysis grouped the active components and functional indexes into three categories. In summary, red beetroot juice fermented by L. acidophilus LA-5 is a source of high-quality antioxidants, as well asAChE andACE inhibitors dominated by betacyanins, which prevent the development of Alzheimer's disease. [ABSTRACT FROM AUTHOR]