Characterization of a Novel Esterase Est33 From an Antarctic Bacterium: A Representative of a New Esterase Family

• Published in: Frontiers in Microbiology
• Main Authors: Liu, Xiaoyu, Zhou, Mingyang, Sun, Rui, Xing, Shu, Wu, Tao, He, Hailun, Chen, Jianbin, Bielicki, John Kevin
• Other Authors: National Natural Science Foundation of China, Natural Science Foundation of Shandong Province, Shandong Academy of Sciences
• Format: Article in Journal/Newspaper
• Language: unknown
• Published: Frontiers Media SA 2022
• Subjects: Microbiology (medical); Microbiology; Antarctic; Antarc*
• Online Access: http://dx.doi.org/10.3389/fmicb.2022.855658; https://www.frontiersin.org/articles/10.3389/fmicb.2022.855658/full

Studies of microorganisms from extreme environments can sometimes reveal novel proteins with unique properties. Here, we identified a novel esterase gene ( Est33 ) from an Antarctic bacterium. The protein was expressed and purified for biochemical characterizations. Site-mutation variants including S94A, D205A, and H233A were constructed to explore the structure–function relationship of the catalytic triad of Est33, and we found mutating Ser 94 , Asp 205 , and His 233 residues lead to a complete loss of enzyme activity. In addition, the catalytic Ser 94 located in a conserved pentapeptide motif GVSWG. Phylogenetic analysis showed that Est33 and its closely related homologs belonged to an independent group apart from other known family members, indicating that Est33 represented a new family of esterase. The Est33 enzyme was found to be a cold-active esterase retaining 25%–100% activity from 10°C to 30°C and to have optimal catalytic activity toward p -nitrophenol acetate (30°C and pH7.5). The serine modifying reagent phenylmethylsulfonyl fluoride inhibited the activity of Est33 by 77.34%, while thiol reagents such as dithiol threitol (DTT) activated the enzyme by 3-fold. Metal chelating reagents EDTA had no effects, indicating that Est33 is not a metalloenzyme. Collectively, these results indicate that Est33 constitutes the first member of a novel esterase family XXI that has been identified.