caa a22 4500 605498083 CHVBK 20210128100544.0 cr unu---uuuuu 210128e20150101xx s 000 0 eng 10.1007/s00253-014-5896-z doi (NATIONALLICENCE)springer-10.1007/s00253-014-5896-z Molecular characterization of a novel thermal stable reductase capable of decoloration of both azo and triphenylmethane dyes [Elektronische Daten] [Fen Gao, Haitao Ding, Lida Shao, Xiaohong Xu, Yuhua Zhao] The gene encoding a putative triphenylmethane reductase (TMR)-like protein derived from Geobacillus thermoglucosidasius C56-Y593 (named as GtAZR) was synthesized, heterologously expressed in Escherichia coli, and extensively characterized for the first time. The recombinant GtAZR displayed its maximum activity at pH5.5 and 40°C. GtAZR was stable at temperatures below 65°C. It also exhibited a broad pH stability and retained more than 90% of its initial activities in pH range of 4.5-10.5 after incubating in various buffers for 1h. Moreover, GtAZR showed significant stability against metal ions and organic solvents. GtAZR displayed broad substrate spectrum toward both azo and triphenylmethane dyes. As a sequence and structural TMR-like protein, GtAZR was characterized as an azoreductase biochemically due to its high specificity for azo dye rather than triphenylmethane dye. Molecular docking and mutagenesis analysis revealed that amino acids Asp-79 and Thr-80 are responsible for its azoreductase activity, which eliminated the steric hindrance caused by His-77 and Tyr-78 at the correspond sites in other structural homologous triphenylmethane reductase. The robust stability and substrate promiscuity of GtAZR made it a promising candidate for practical removal of mixed dye wastewater. Springer-Verlag Berlin Heidelberg, 2014 Geobacillus thermoglucosidasius nationallicence Azoreductase nationallicence Triphenylmethane reductase nationallicence Dye degradation nationallicence Stability nationallicence Substrate promiscuity nationallicence Gao Fen Institute of Microbiology, College of Life Science, Zhejiang University, 310058, Hangzhou, China aut Ding Haitao Institute of Microbiology, College of Life Science, Zhejiang University, 310058, Hangzhou, China aut Shao Lida Department of Horticulture, College of Agriculture and Biotechnology, Zhejiang University, 310058, Hangzhou, China aut Xu Xiaohong Hangzhou Sunzymes Biotechnology Co., Ltd., 310013, Hangzhou, China aut Zhao Yuhua Institute of Microbiology, College of Life Science, Zhejiang University, 310058, Hangzhou, China aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/1(2015-01-01), 255-267 0175-7598 99:1<255 2015 99 253 https://doi.org/10.1007/s00253-014-5896-z text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-014-5896-z text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Gao Fen Institute of Microbiology, College of Life Science, Zhejiang University, 310058, Hangzhou, China aut NATIONALLICENCE 700 1- Ding Haitao Institute of Microbiology, College of Life Science, Zhejiang University, 310058, Hangzhou, China aut NATIONALLICENCE 700 1- Shao Lida Department of Horticulture, College of Agriculture and Biotechnology, Zhejiang University, 310058, Hangzhou, China aut NATIONALLICENCE 700 1- Xu Xiaohong Hangzhou Sunzymes Biotechnology Co., Ltd., 310013, Hangzhou, China aut NATIONALLICENCE 700 1- Zhao Yuhua Institute of Microbiology, College of Life Science, Zhejiang University, 310058, Hangzhou, China aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/1(2015-01-01), 255-267 0175-7598 99:1<255 2015 99 253