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Characterization of the second and third steps in the enzymatic pathway for microcystin-RR biodegradation by Sphingopyxis sp. USTB-05

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Abstract

Microcystin-RR (MC-RR) is a dominant variant of microcystin (MC) worldwide. We previously isolated Sphingopyxis sp. USTB-05, which can efficiently biodegrade MC-RR. At least three enzymes encoded by MC-degrading genes are involved. Based on the successful expression of the first gene USTB-05-A, the second (USTB-05-B, KC513423) and third (USTB-05-C, KC573527) genes involved in the biodegradation of MC-RR were further cloned from Sphingopyxis sp. USTB-05 and expressed in Escherichia coli BL21 (DE3). After purification, the MC-degrading enzymes encoded by these genes were used for the catalytic degradation of MC-RR. The results demonstrated that the second enzyme encoded by USTB-05-B could convert linear MC-RR to a tetrapeptide by breaking the Ala–Arg bond. The third enzyme encoded by USTB-05-C could cleave Adda-Glu peptide bonds of both linear MC-RR and the tetrapeptide of Adda-Glu-Mdha-Ala, producing Adda as their common product. These findings will help better understand the biodegradation mechanism of MCs by Sphingopyxis sp. USTB-05.

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Acknowledgments

This work was supported jointly by the National Natural Science Foundation of China (No. 21177009 and No. 11071013), Specific Foundation of Doctoral program, Ministry of Education, P.R. China (No.20120006110001) and Program of International S & T Cooperation, Chinese MOST (2010DFA92090).

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Correspondence to Hai Yan.

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Wang, H., Yan, H., Ma, S. et al. Characterization of the second and third steps in the enzymatic pathway for microcystin-RR biodegradation by Sphingopyxis sp. USTB-05. Ann Microbiol 65, 495–502 (2015). https://doi.org/10.1007/s13213-014-0885-0

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