AI Article Synopsis

  • - The study focuses on a specific β-1,3-glucan synthase (CMGLS) in fungi, highlighting its crucial role in glucan synthesis and cell wall formation despite challenges posed by its complex structure and size.
  • - Researchers successfully cloned the gene for CMGLS, partially purified the enzyme, and determined its specific activity, binding specificity to UDP-glucose, and maximum degree of polymerization.
  • - Utilizing AlphaFold and molecular docking, the 3D structure of CMGLS was analyzed, revealing important interactions between UDP-glucose and the enzyme, which enhances understanding of glucan synthesis mechanisms in fungi.

Article Abstract

β-1,3-Glucan synthases play key roles in glucan synthesis, cell wall assembly, and growth of fungi. However, their multi-transmembrane domains (over 14 TMHs) and large molecular masses (over 100 kDa) significantly hamper understanding of their catalytic characteristics and mechanisms. In the present study, the 5841-bp gene encoding the 221.7 kDa membrane-bound β-1,3-glucan synthase CMGLS in was cloned, identified, and structurally analyzed. CMGLS was partially purified with a specific activity of 87.72 pmol/min/μg, a purification fold of 121, and a yield of 10.16% using a product-entrapment purification method. CMGLS showed a strict specificity to UDP-glucose with a value of 84.28 μM at pH 7.0 and synthesized β-1,3-glucan with a maximum degree of polymerization (DP) of 70. With the assistance of AlphaFold and molecular docking, the 3D structure of CMGLS and its binding features with substrate UDP-glucose were proposed for the first time to our knowledge. UDP-glucose potentially bound to at least 11 residues hydrogen bonds, π-stacking ,and salt bridges, and Arg 1436 was predicted as a key residue directly interacting with the moieties of glucose, phosphate, and the ribose ring on UDP-glucose. These findings would open an avenue to recognize and understand the glucan synthesis process and catalytic mechanism of β-1,3-glucan synthases in mushrooms.

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http://dx.doi.org/10.1021/acs.jafc.2c03410DOI Listing

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