5GM5

Crystal structure of FI-CMCase from Aspergillus aculeatus F-50 in complex with cellobiose

Summary for 5GM5

• Entry DOI: 10.2210/pdb5gm5/pdb
• Related: 5GM3 5GM4
• Related PRD ID: PRD_900005
• Descriptor: Endoglucanase-1, beta-D-glucopyranose-(1-4)-beta-D-glucopyranose, SULFATE ION, ... (5 entities in total)
• Functional Keywords: substrate binding, hydrolase-inhibitor complex, hydrolase/inhibitor
• Biological source: Aspergillus aculeatus
• Cellular location: Secreted: P22669
• Total number of polymer chains: 7
• Total formula weight: 169414.90

Authors

Huang, J.W.,Liu, W.D.,Zheng, Y.Y.,Chen, C.C.,Guo, R.T. (deposition date: 2016-07-12, release date: 2017-05-17, Last modification date: 2024-10-16)

Primary citation

Huang, J.W.,Liu, W.,Lai, H.L.,Cheng, Y.S.,Zheng, Y.,Li, Q.,Sun, H.,Kuo, C.J.,Guo, R.T.,Chen, C.C.
Crystal structure and genetic modifications of FI-CMCase from Aspergillus aculeatus F-50
Biochem. Biophys. Res. Commun., 478:565-572, 2016

PubMed Abstract: Cellulose is the major component of the plant cell wall and the most abundant renewable biomass on earth, and its decomposition has proven to be very useful in many commercial applications. Endo-1,4-β-d-glucanase (EC 3.2.1.4; endoglucanase), which catalyzes the random hydrolysis of 1,4-β-glycosidic bonds of the cellulose main chain to cleave cellulose into smaller fragments, is the key cellulolytic enzyme. An endoglucanase isolated from Aspergillus aculeatus F-50 (FI-CMCase), which is classified into the glycoside hydrolase (GH) family 12, was demonstrated to be effectively expressed in the industrial strain Pichia pastoris. Here, the crystal structure and complex structures of P. pastoris-expressed FI-CMCase were solved to high resolution. The overall structure is analyzed and compared to other GH12 members. In addition, the substrate-surrounding residues were engineered to search for variants with improved enzymatic activity. Among 14 mutants constructed, one with two-fold increase in protein expression was identified, which possesses a potential to be further developed as a commercial enzyme product.

PubMed: 27470581
DOI: 10.1016/j.bbrc.2016.07.101

Experimental method

X-RAY DIFFRACTION (1.73 Å)