Accumulation of non-degradable plastic waste in the environment might be prevented by the use of biodegradable polyhydroxyalkanoate (PHA). In this study, the thermophile Schlegelella thermodepolymerans produced up to 80 wt% PHA based on dry cell mass. The largest PHA granules were found in the cells within 48 h using 20 g/L xylose, a C/N ratio of 100, an initial pH of 7, at 50 °C. The substrate consumption, pH changes, and cell growth were monitored, revealing the time dependency of PHA production in S. thermodepolymerans. The metabolic pathways from xylose to PHA were identified based on proteomic analysis, revealing involvement of classic phaCAB, de novo fatty acid biosynthesis, and fatty acid β-oxidation. In addition, it was shown that S. thermodepolymerans degraded extracellular PHA with a high efficiency at 50 °C.