Despite numerous efforts to discover novel bioactive products from microorganisms, previously reported compounds are repetitively reisolated. A new polyketide glycoside, gordonic acid (1), isolated from the mixed culture of two Gram-positive bacteria, Gordonia sp. KMC005 and Streptomyces tendae KMC006, is reported. The structure of 1 was characterized as an acyclic polyene polyketide substituted with a β-d-digitoxopyranose through NMR, HR-ESI-QTOF-MS, IR, and UV spectral data. The stereochemistry for 1 was determined by Mosher's method followed by 2D NOESY analysis and by NMR chemical shift calculations supported by DP4 analysis. Gordonic acid (1) showed weak activity against Micrococcus luteus and Enterococcus hirae.