Do_2022_Crystals_12_46

Reference

Title : Sequence Analysis and Preliminary X-ray Crystallographic Analysis of an Acetylesterase (LgEstI) from Lactococcus garvieae - Do_2022_Crystals_12_46
Author(s) : Do H , Wang Y , Lee CW , Yoo W , Jeon S , Hwang J , Lee MJ , Kim KK , Kim HW , Lee JH , Kim TD
Ref : Crystals , 12 :46 , 2022
Abstract : A gene encoding LgEstI was cloned from a bacterial fish pathogen, Lactococcus garvieae. Sequence and bioinformatic analysis revealed that LgEstI is close to the acetyl esterase family and had maximum similarity to a hydrolase (UniProt: Q5UQ83) from Acanthamoeba polyphaga mimivirus (APMV). Here, we present the results of LgEstI overexpression and purification, and its preliminary X-ray crystallographic analysis. The wild-type LgEstI protein was overexpressed in Escherichia coli, and its enzymatic activity was tested using p-nitrophenyl of varying lengths. LgEstI protein exhibited higher esterase activity toward p-nitrophenyl acetate. To better understand the mechanism underlying LgEstI activity and subject it to protein engineering, we determined the high-resolution crystal structure of LgEstI. First, the wild-type LgEstI protein was crystallized in 0.1 M Tris-HCl buffer (pH 7.1), 0.2 M calcium acetate hydrate, and 19% (w/v) PEG 3000, and the native X-ray diffraction dataset was collected up to 2.0 A resolution. The crystal structure was successfully determined using a molecular replacement method, and structure refinement and model building are underway. The upcoming complete structural information of LgEstI may elucidate the substrate-binding mechanism and provide novel strategies for subjecting LgEstI to protein engineering.
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Gene_locus related to this paper: 9lact-a0a5m9r5n4

Related information

Gene_locus related to this paper: 9lact-a0a5m9r5n4

Citations formats

Do H, Wang Y, Lee CW, Yoo W, Jeon S, Hwang J, Lee MJ, Kim KK, Kim HW, Lee JH, Kim TD (2022)
Sequence Analysis and Preliminary X-ray Crystallographic Analysis of an Acetylesterase (LgEstI) from Lactococcus garvieae
Crystals 12 :46

Do H, Wang Y, Lee CW, Yoo W, Jeon S, Hwang J, Lee MJ, Kim KK, Kim HW, Lee JH, Kim TD (2022)
Crystals 12 :46