Extremophile bacteria originally isolated from the hot springs of Gilgit Baltistan and Khewara salt mine, i.e., Geobacillus stearothermophilus strain MB600 and Halomonas elongata strain MB591, were tested for their production of extremozymes. The targeted enzyme class was hydrolytic enzymes (Lipase, Gelatinase, Protease, & Cellulase), and isomerase enzymes (aconitase, hydratase, and phosphoglycerate mutase). The thermophilic bacteria MB600 produced all hydrolytic enzymes after 24 hours of incubation. Matrixassisted laser desorption/ionization-time of flight (MALDI TOF) analysis confirmed the presence of Aconitase hydratase isolated from Geobacillus stearothermophilus strain MB600 with a molecular weight of 98 KD. Similarly, all the hydrolytic enzymes were produced by Halomonas elongata strain MB591. The production of Phosphoglycerate mutase by Halomonas elongata strain MB591 was also confirmed, and its molecular characterization was performed using Sodium Dodecyl Sulphate-Polyacrylamide Gel Electrophoresis (SDS-PAGE) and MALDI-TOF with a molecular weight of 56 KD. Later, insilico analysis was also performed for predicting the 3D structure of enzymes, and their bonding interactions with guaiacol were explained by docking using CB Dock.