The β-glucosidases known to improve tea aroma are all mesothermal enzymes, limiting their use under brewing conditions. Based on the properties analysis and molecular docking, the thermostable β-glucosidase (TPG) from Thermotoga petrophlia showed potential to enhance tea aroma. Treatment by recombinant TPG at 90 °C, the floral, sweet and grassy notes of instant Oolong tea were increased, while the roasted, caramel and woody notes were decreased. The improved floral, sweet and grassy notes were related to increase releasing of benzyl alcohol (floral), geraniol (floral), (Z)-3-hexen-1-ol (grassy), benzaldehyde (sweet) and 1-hexanol (grassy) by TPG hydrolyzing of (Z)-3-hexenyl-β-D-glucopyranoside, hexanyl-β-D-glucopyranoside (HGP), benzyl-β-D-glucopyranoside, prunasin and geranyl-β-D-glucopyranoside (GGP), respectively. Although the catalytic efficiency of TGP to GGP was about twice that to HGP, HPG was more competitive than GGP when they mixed. Combined with microstructure analysis, the structure-function relationship of TPG-influencing tea aroma were understood. This study provided the method of how to mining new function of characterized β-glucosidases, as well as a theoretical basis for the development of new tea products.