Improving Both the Thermostability and Catalytic Efficiency of Phospholipase D from Moritella sp. JT01 through Disulfide Bond Engineering Strategy.

ABSTRACT: Mining of Phospholipase D (PLD) with high activity and stability has attracted strong interest for investigation. A novel PLD from marine Moritella sp. JT01 (MsPLD) was biochemically and structurally characterized in our previous study; however, the short half-life time (t_1/2) under its optimum reaction temperature seriously hampered its further applications. Herein, the disulfide bond engineering strategy was applied to improve its thermostability. Compared with wild-type MsPLD, mutant S148C-T206C/D225C-A328C with the addition of two disulfide bonds exhibited a 3.1-fold t_1/2 at 35 °C and a 5.7 °C increase in melting temperature (T_m). Unexpectedly, its specific activity and catalytic efficiency (k_cat/K_m<

SUBMITTER: Li L 

PROVIDER: S-EPMC9570233 | biostudies-literature | 2022 Sep

REPOSITORIES: biostudies-literature