Schizosaccharomyces pombe YM-19 and Acetobacter pasteurianus EM2-03 were isolated and identified from the traditional herbal vinegar fermentation process using cultural and molecular techniques. The two microorganisms were prepared to inoculum for an experimental herbal vinegar fermentation, which is called “Loog Plaag Mea.” The first starter culture was 10% v/v S. pombe YM1-19. Fermentation was carried out under anaerobic conditions at 30°C for 4 days. The alcohol content was 6.18±0.13% v/v and the pH value was 3.52±0.02 on day 4 of the fermentation period. Subsequently, 10% v/v A. pasteurianus EM2-03 was added to the fermentation process under aerobic conditions at 30°C for 22 days. The final herbal vinegar product contained 4.91±0.15% v/v acetic acid, and a pH value of 3.04±0.04. Its total phenolic content and IC50 value of DPPH radical scavenging were 1,908.38±38.75 µgGAE/mL and 0.017±0.001 µL/mL, respectively. The experimental fermented herbal vinegar had physicochemical properties that very similar to traditional vinegar. As a result, the isolated microorganisms can be used to improve product consistency and quality control in the mass production of vinegar.