(Below N is a link to NCBI taxonomic web page and E link to ESTHER at designed phylum.) > cellular organisms: NE > Bacteria: NE > Terrabacteria group: NE > Firmicutes: NE > Bacilli: NE > Lactobacillales: NE > Streptococcaceae: NE > Lactococcus: NE > Lactococcus garvieae: NE
Warning: This entry is a compilation of different species or line or strain with more than 90% amino acide identity. You can retrieve all strain data
(Below N is a link to NCBI taxonomic web page and E link to ESTHER at designed phylum.) Lactococcus garvieae DCC43: N, E.
Lactococcus garvieae TRF1: N, E.
Lactococcus garvieae ATCC 49156: N, E.
Lactococcus garvieae Lg2: N, E.
LegendThis sequence has been compared to family alignement (MSA) red => minority aminoacid blue => majority aminoacid color intensity => conservation rate title => sequence position(MSA position)aminoacid rate Catalytic site Catalytic site in the MSA MVERISLEKAALEFSEANAPHPRIYELPVEEGRSLLNEVQDSPVVKEDVD IEDIAVDTGEWGEINVRFIRPLHQEKKLPVIFYIHGAGWVFGNAHTHDKL IRELAVRTNSVVVFSEYSLSPEAKYPTAIEQNYAVLQQLKDFANDKKFDV NHLTVAGDSVGGNMATVMTLLTKQRGGQKIGQQVLYYPVTDANFDTDSYN EFAENYFLTKEGMIWFWDQYTTSQEERHQITASPLRATKEDLADLPAALI ITGEADVLRDEGEAYARKLREADVEVTQVRFQAIIHDFVMVNSMNETHAT RAAMSLSTQWINEKNRK
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.
Lactococcus garvieae Acetylesterase (LgEstI)
