Hyaluronate lyases are widely used in medicine and biochemical engineering and are also applied as a tool enzyme to prepare oligosaccharides with various biological activities. To date, only a few hyaluronate lyases are on sale with poor thermostability. In this study, a PL8 hyaluronate lyase, TcHly8D, was found from and expressed in with a maximum yield of 1.77 × 10 U/L (3.14 g/L) in a 5-L bioreactor. The recombinant TcHly8D exhibited a high hyaluronate lyase activity of 5.64 × 10 U/mg and an excellent thermostability with half-lives of 184.9 h at 60 °C. Fifty micrograms of TcHly8D could catalyze 5 g of hyaluronic acid with an oligosaccharide yield of 84.8% in 4 h. The salt bridges, hydrogen bonds, and proline residues, but not disulfide bonds, played important roles in the thermostability of TcHly8D. These findings provide insights into the multifunctional application potential of TcHly8D in agriculture, medicine, and the food industry.