title: Structure-based rational design to enhance the solubility and thermostability of a bacterial laccase Lac15.
abstract: Bacterial laccases are ideal alternatives of fungal laccases for specific industrial applications due to specific characteristics such as alkalescence dependence and high chloride tolerance. However, some bacterial laccases presented as inclusion bodies when expressing in Escherichia coli and showed thermal instability. In this study, rational design was employed to enhance the solubility and the thermostablity of the bacterial laccase Lac15-His6 based on the crystal structure obtained previously. After deletion of His-tag and residues323-332, the obtained Lac15D was completely expressed in soluble form even at a higher temperature of 28°C, compared to only 50% of Lac15-His6 expressed solubly at 16°C. It showed a two-time higher activity at temperatures lower than 35°C and a half-life increasing from 72 min to 150 min at 45°C. When used in chromogenic reactions, Lac15D showed constant activity toward dye precursors and their combinations under alkaline conditions, demonstrating its application potential in hair coloring biotechnology.
authors.affiliation: School of Life Sciences, Anhui University, Hefei, Anhui, China; Anhui Provincial Engineering Technology Research Center of Microorganisms and Biocatalysis, Hefei, Anhui, China.
authors.forename: Zemin
authors.initials: Z
authors.lastname: Fang
authors.affiliation: School of Life Sciences, Anhui University, Hefei, Anhui, China; Anhui Provincial Engineering Technology Research Center of Microorganisms and Biocatalysis, Hefei, Anhui, China.
authors.forename: Peng
authors.initials: P
authors.lastname: Zhou
authors.affiliation: School of Life Sciences, Anhui University, Hefei, Anhui, China; Anhui Provincial Engineering Technology Research Center of Microorganisms and Biocatalysis, Hefei, Anhui, China.
authors.forename: Fei
authors.initials: F
authors.lastname: Chang
authors.affiliation: School of Life Sciences, Anhui University, Hefei, Anhui, China; Anhui Provincial Engineering Technology Research Center of Microorganisms and Biocatalysis, Hefei, Anhui, China.
authors.forename: Qiang
authors.initials: Q
authors.lastname: Yin
authors.affiliation: School of Life Sciences, Anhui University, Hefei, Anhui, China; Anhui Provincial Engineering Technology Research Center of Microorganisms and Biocatalysis, Hefei, Anhui, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Fang
authors.affiliation: School of Life Sciences, Anhui University, Hefei, Anhui, China; Anhui Provincial Engineering Technology Research Center of Microorganisms and Biocatalysis, Hefei, Anhui, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Yuan
authors.affiliation: School of Life Sciences, Anhui University, Hefei, Anhui, China; Anhui Provincial Engineering Technology Research Center of Microorganisms and Biocatalysis, Hefei, Anhui, China.
authors.forename: Xuecheng
authors.initials: X
authors.lastname: Zhang
authors.affiliation: School of Life Sciences, Anhui University, Hefei, Anhui, China; Anhui Provincial Engineering Technology Research Center of Microorganisms and Biocatalysis, Hefei, Anhui, China.
authors.forename: Yazhong
authors.initials: Y
authors.lastname: Xiao
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry*; D002384:Catalysis; D002863:Chromogenic Compounds / Q000378:metabolism; D018360:Crystallography, X-Ray; D004926:Escherichia coli; D006202:Hair Dyes / Q000378:metabolism; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D042845:Laccase / Q000737:chemistry*; D008958:Models, Molecular; D011487:Protein Conformation; D015202:Protein Engineering; D055550:Protein Stability; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D012995:Solubility; D013329:Structure-Activity Relationship; D013379:Substrate Specificity
chemical_list: D001426:Bacterial Proteins; D002863:Chromogenic Compounds; D006202:Hair Dyes; D011993:Recombinant Fusion Proteins; D042845:Laccase