Crystal Structure of GH49 Dextranase from Arthrobacter oxidans KQ11: Identification of Catalytic Base and Improvement of Thermostability Using Semirational Design Based on B‑Factors
journal contributionposted on 28.03.2019, 00:00 by Wei Ren, Le Liu, Lide Gu, Wanli Yan, Yan li Feng, Dongxue Dong, Shujun Wang, Mingsheng Lyu, Changhai Wang
The crystal structure of Dextranase from the marine bacterium Arthrobacter oxidans KQ11 (Aodex) was determined at a resolution of 1.4 Å. The crystal structure of the conserved Aodex fragment (Ala52–Thr638) consisted of an N-terminal domain N and a C-terminal domain C. The N-terminal domain N was identified as a β-sandwich, connected to a right-handed parallel β-helix at the C-terminus. Sequence comparisons, cavity regions, and key residues of the catalytic domain analysis all suggested that the Aodex was an inverting enzyme, and the catalytic acid and base were Asp439 and Asp420, respectively. Asp440 was not a general base in the Aodex catalytic domain, and Asp396 in Dex49A may not be a general base in the catalytic domain. The thermostability of the S357F mutant using semirational design based on B-factors was clearly better than that of wild-type Aodex. This process may promote the aromatic–aromatic interactions that increase the thermostability of mutant Phe357.