Crystal structure of a thermophilic fungal cyanase and its implications on the catalytic mechanism for bioremediation

Ranjan, Bibhuti; Choi, Philip H.; Pillai, Santhosh; Permaul, Kugenthiren; Tong, Liang; Singh, Suren

doi:10.1038/s41598-020-79489-3

Crystal structure of a thermophilic fungal cyanase and its implications on the catalytic mechanism for bioremediation

Journal Article · Mon Jan 11 00:00:00 EST 2021 · Scientific Reports

DOI:https://doi.org/10.1038/s41598-020-79489-3· OSTI ID:1765222

Ranjan, Bibhuti ^[1]; Choi, Philip H. ^[2]; Pillai, Santhosh ^[3]; Permaul, Kugenthiren ^[3]; Tong, Liang ^[1]; Singh, Suren ^[3]

Durban Univ. of Technology (South Africa); Columbia Univ., New York, NY (United States)
Columbia Univ., New York, NY (United States)
Durban Univ. of Technology (South Africa)

Cyanase catalyzes the bicarbonate-dependent degradation of cyanate to produce ammonia and carbon dioxide, and ammonia is a considerable alternative nitrogen source. Strikingly, the cyanase from the thermophilic fungus Thermomyces lanuginosus (Tl-Cyn) has the highest catalytic efficiency reported among these enzymes. However, its molecular mechanism of action is not clearly understood, because currently there is no structural information available on fungal cyanases. Here we report the crystal structure of Tl-Cyn in complex with inhibitors malonate and formate at 2.2 Å resolution. The structure reveals extensive interactions at the subunit interfaces in a dimer, and a decamer is formed by a pentamer of these dimers. Our biochemical, kinetic and mutagenesis studies confirm the structural observations on the complex and provide further insights into its catalytic mechanism and inhibition. The structure has also aided the creation of a mutant enzyme with enhanced catalytic activity, and such enzymes may have the potential for biotechnological applications, including biotransformation and bioremediation. Moreover, other fungal cyanases with potentially high catalytic activity could also be predicted based on the Tl-Cyn structure, as the active site region among fungal cyanases are highly conserved.

View Accepted Manuscript (DOE)

Research Organization:: Argonne National Laboratory (ANL), Argonne, IL (United States). Advanced Photon Source (APS)

Sponsoring Organization:: USDOE Office of Science (SC)

Grant/Contract Number:: AC02-06CH11357

OSTI ID:: 1765222

Alternate ID(s):: OSTI ID: 1783936

Journal Information:: Scientific Reports, Journal Name: Scientific Reports Journal Issue: 1 Vol. 11; ISSN 2045-2322

Publisher:: Nature Publishing GroupCopyright Statement

Country of Publication:: United States

Language:: ENGLISH

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