High hydrostatic pressure (HHP) stabilized glucose oxidase (GOx) against thermal inactivation. The apparent first-order kinetics of inactivation of GOx were investigated at 0.1-300MPa and 58.8-80.0 degrees C. At 240MPa and 74.5 degrees C, GOx inactivated at a rate 50 times slower than at atmospheric pressure at the same temperature. The apparent activation energy of inactivation at 300MPa was 281.0 +/- 17.4kJmol(-1) or 1.3-fold smaller than for the inactivation at atmospheric pressure (378.1 +/- 25.6kJmol(-1)). The stabilizing effect of HHP was greatest at 74.5 degrees C, where the activation volume of 57.0 +/- 12.0cm(3)mol(-1) was highest compared to all other studied temperatures. Positive apparent activation volumes for all the treatment temperatures confirmed that HHP favors GOx stabilization. A second approach to increase GOx stability involved crosslinking with N-(3-dimethylaminopropyl)-N-ethylcarbodiimide hydrochloride (EDC) and either aniline or benzoate. The modified enzyme remained fully active with only slight increases in K-M (1.3-1.9-fold increases for aniline and benzoate modification, respectively). The thermal stability of GOx increased by 8 degrees C with aniline modification, while it decreased by 0.9 degrees C upon modification with benzoate. Biotechnol. Bioeng. 2017;114: 516-525. (c) 2016 Wiley Periodicals, Inc.