Engineering the Catalytic Properties of Two-Domain Laccase from Ac-993

Overview

Journal Int J Mol Sci

Publisher MDPI

Specialties Biochemistry
Chemistry
Molecular Biology

Date 2022 Jan 11

PMID 35008493

Authors

Ilya Kolyadenko

Anastasia Scherbakova

Kirill Kovalev

Azat Gabdulkhakov

Svetlana Tishchenko

Affiliations

Soon will be listed here.

Abstract

Laccases catalyze the oxidation of substrates with the concomitant reduction of oxygen to water. Recently, we found that polar residues located in tunnels leading to Cu2 and Cu3 ions control oxygen entrance (His 165) and proton transport (Arg 240) of two-domain laccase (2D) from (SgfSL). In this work, we have focused on optimizing the substrate-binding pocket (SBP) of SgfSL while simultaneously adjusting the oxygen reduction process. SgfSL variants with three single (Met199Ala, Met199Gly, and Tyr230Ala) and three double amino acid residues substitutions (Met199Gly/His165Ala, His165Ala/Arg240His, Met199Gly/Arg240His) were constructed, purified, and investigated. Combination of substitutions in the SBP and in the tunnel leading to Cu2 ion (Met199Gly/Arg240His) increased SgfSL catalytic activity towards ABTS by 5-fold, and towards 2.6-DMP by 16-fold. The high activity of the Met199Gly/Arg240His variant can be explained by the combined effect of the SBP geometry optimization (Met199Gly) and increased proton flux via the tunnel leading to Cu2 ion (Arg240His). Moreover, the variant with Met199Gly and His165Ala mutations did not significantly increase SgfSL's activity, but led to a drastic shift in the optimal pH of 2.6-DMP oxidation. These results indicate that His 165 not only regulates oxygen access, but it also participates in proton transport in 2D laccases.

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References

Murshudov G, Skubak P, Lebedev A, Pannu N, Steiner R, Nicholls R . REFMAC5 for the refinement of macromolecular crystal structures. Acta Crystallogr D Biol Crystallogr. 2011; 67(Pt 4):355-67. PMC: 3069751. DOI: 10.1107/S0907444911001314. View

Heinfling A, Martinez M, Martinez A, Bergbauer M, Szewzyk U . Purification and characterization of peroxidases from the dye-decolorizing fungus Bjerkandera adusta. FEMS Microbiol Lett. 1998; 165(1):43-50. DOI: 10.1111/j.1574-6968.1998.tb13125.x. View

Lu L, Zhao M, Wang T, Zhao L, Du M, Li T . Characterization and dye decolorization ability of an alkaline resistant and organic solvents tolerant laccase from Bacillus licheniformis LS04. Bioresour Technol. 2011; 115:35-40. DOI: 10.1016/j.biortech.2011.07.111. View

Machczynski M, Vijgenboom E, Samyn B, Canters G . Characterization of SLAC: a small laccase from Streptomyces coelicolor with unprecedented activity. Protein Sci. 2004; 13(9):2388-97. PMC: 2280001. DOI: 10.1110/ps.04759104. View

Chen Z, Durao P, Silva C, Pereira M, Todorovic S, Hildebrandt P . The role of Glu498 in the dioxygen reactivity of CotA-laccase from Bacillus subtilis. Dalton Trans. 2010; 39(11):2875-82. DOI: 10.1039/b922734b. View

Heinzkill M, Bech L, Halkier T, Schneider P, Anke T . Characterization of laccases and peroxidases from wood-rotting fungi (family Coprinaceae). Appl Environ Microbiol. 1998; 64(5):1601-6. PMC: 106202. DOI: 10.1128/AEM.64.5.1601-1606.1998. View

Gunne M, Urlacher V . Characterization of the alkaline laccase Ssl1 from Streptomyces sviceus with unusual properties discovered by genome mining. PLoS One. 2013; 7(12):e52360. PMC: 3527528. DOI: 10.1371/journal.pone.0052360. View

Gunne M, Hoppner A, Hagedoorn P, Urlacher V . Structural and redox properties of the small laccase Ssl1 from Streptomyces sviceus. FEBS J. 2014; 281(18):4307-18. DOI: 10.1111/febs.12755. View

Kallio J, Auer S, Janis J, Andberg M, Kruus K, Rouvinen J . Structure-function studies of a Melanocarpus albomyces laccase suggest a pathway for oxidation of phenolic compounds. J Mol Biol. 2009; 392(4):895-909. DOI: 10.1016/j.jmb.2009.06.053. View

10.

Vandeyar M, Weiner M, HUTTON C, Batt C . A simple and rapid method for the selection of oligodeoxynucleotide-directed mutants. Gene. 1988; 65(1):129-33. DOI: 10.1016/0378-1119(88)90425-8. View

11.

Wariishi H, Valli K, Gold M . Manganese(II) oxidation by manganese peroxidase from the basidiomycete Phanerochaete chrysosporium. Kinetic mechanism and role of chelators. J Biol Chem. 1992; 267(33):23688-95. View

12.

Giardina P, Faraco V, Pezzella C, Piscitelli A, Vanhulle S, Sannia G . Laccases: a never-ending story. Cell Mol Life Sci. 2009; 67(3):369-85. PMC: 11115910. DOI: 10.1007/s00018-009-0169-1. View

13.

Silva C, Damas J, Chen Z, Brissos V, Martins L, Soares C . The role of Asp116 in the reductive cleavage of dioxygen to water in CotA laccase: assistance during the proton-transfer mechanism. Acta Crystallogr D Biol Crystallogr. 2012; 68(Pt 2):186-93. DOI: 10.1107/S0907444911054503. View

14.

Jones S, Solomon E . Electron transfer and reaction mechanism of laccases. Cell Mol Life Sci. 2015; 72(5):869-83. PMC: 4323859. DOI: 10.1007/s00018-014-1826-6. View

15.

Pan K, Zhao N, Yin Q, Zhang T, Xu X, Fang W . Induction of a laccase Lcc9 from Coprinopsis cinerea by fungal coculture and its application on indigo dye decolorization. Bioresour Technol. 2014; 162:45-52. DOI: 10.1016/j.biortech.2014.03.116. View

16.

Osipov E, Polyakov K, Kittl R, Shleev S, Dorovatovsky P, Tikhonova T . Effect of the L499M mutation of the ascomycetous Botrytis aclada laccase on redox potential and catalytic properties. Acta Crystallogr D Biol Crystallogr. 2014; 70(Pt 11):2913-23. PMC: 4220974. DOI: 10.1107/S1399004714020380. View

17.

DArcy A, Bergfors T, Cowan-Jacob S, Marsh M . Microseed matrix screening for optimization in protein crystallization: what have we learned?. Acta Crystallogr F Struct Biol Commun. 2014; 70(Pt 9):1117-26. PMC: 4157405. DOI: 10.1107/S2053230X14015507. View

18.

Yin Q, Zhou G, Peng C, Zhang Y, Kues U, Liu J . The first fungal laccase with an alkaline pH optimum obtained by directed evolution and its application in indigo dye decolorization. AMB Express. 2019; 9(1):151. PMC: 6751238. DOI: 10.1186/s13568-019-0878-2. View

19.

El-Bendary M, Ezzat S, Ewais E, Al-Zalama M . Optimization of spore laccase production by isolated from wastewater and its potential in green biodecolorization of synthetic textile dyes. Prep Biochem Biotechnol. 2020; 51(1):16-27. DOI: 10.1080/10826068.2020.1786698. View

20.

Trubitsina L, Tishchenko S, Gabdulkhakov A, Lisov A, Zakharova M, Leontievsky A . Structural and functional characterization of two-domain laccase from Streptomyces viridochromogenes. Biochimie. 2015; 112:151-9. DOI: 10.1016/j.biochi.2015.03.005. View