Dual Activity BLEG-1 from Bacillus lehensis G1 Revealed Structural Resemblance to B3 Metallo-β-Lactamase and Glyoxalase II: An Insight into Its Enzyme Promiscuity and Evolutionary Divergence.

Raja Noor Zaliha Raja Abdul Rahman,Noor Dina Muhd Noor,Shaw Xian Au,Yahaya M Normi,Hiroyoshi Matsumura,Nur Syazana Dzulkifly

doi:10.3390/ijms22179377

Raja Noor Zaliha Raja Abdul Rahman, Noor Dina Muhd Noor + Show 4 more

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https://doi.org/10.3390/ijms22179377

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Abstract

Metallo-β-lactamases (MBLs) are class B β-lactamases from the metallo-hydrolase-like MBL-fold superfamily which act on a broad range of β-lactam antibiotics. A previous study on BLEG-1 (formerly called Bleg1_2437), a hypothetical protein from Bacillus lehensis G1, revealed sequence similarity and activity to B3 subclass MBLs, despite its evolutionary divergence from these enzymes. Its relatedness to glyoxalase II (GLXII) raises the possibility of its enzymatic promiscuity and unique structural features compared to other MBLs and GLXIIs. This present study highlights that BLEG-1 possessed both MBL and GLXII activities with similar catalytic efficiencies. Its crystal structure revealed highly similar active site configuration to YcbL and GloB GLXIIs from Salmonella enterica, and L1 B3 MBL from Stenotrophomonas maltophilia. However, different from GLXIIs, BLEG-1 has an insertion of an active-site loop, forming a binding cavity similar to B3 MBL at the N-terminal region. We propose that BLEG-1 could possibly have evolved from GLXII and adopted MBL activity through this insertion.

Highlights

Metallo-hydrolase-like MBL-fold superfamily of proteins is a large and ancient group of proteins that are widely distributed over the kingdoms of bacteria, archaea, and eukarya
Our results showed that the binding mode of ampicillin in BLEG-1 and L1 are similar in which the aromatic side chain of ampicillin (i.e., 2-amino-2-phenylacetamido group) and penam ring are mainly accommodated by the active site loops in the N- and C-terminal domain respectively (Figure 10A,B)
We demonstrated that BLEG-1 possessed dual MBL and glyoxalase II (GLXII) activities with similar catalytic efficiencies

Summary

Introduction

Metallo-hydrolase-like MBL-fold superfamily of proteins is a large and ancient group of proteins that are widely distributed over the kingdoms of bacteria, archaea, and eukarya. II (GLXII), N-acyl-L-homoserine lactonase (AHL), persulfide dioxygenase, flavodiiron protein, choline-binding protein, cleavage and polyadenylation specificity factors, arylsulfatase, 50 -exonuclease, ribonuclease, cyclic nucleotide phosphodiesterase, insecticide hydrolase, and proteins required for natural transformation competence [1,2,3,4]. Members of this superfamily contain conserved αβ/βα metallo-β-lactamase-fold (MBL-fold) domain, with two central β-sheets surrounded by solvent-exposed α-helices, and either have a mononuclear or binuclear metal binding site located at the interface of the β/β sandwich, Int. J.

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