Functional characterization of a putative β-lactamase gene in the genome of Zymomonas mobilis

Rajnish, K. Narayanan; Asraf, Sheik; Manju, Nagarajan; Gunasekaran, Paramasamy

doi:10.1007/s10529-011-0704-7

Cited by 4 publications

(5 citation statements)

References 18 publications

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“…Both ORFs contained ATG as start codon, which was preceded by a putative ribosome binding site (rbs). The identified rbs for end 1 and 2 matched with reported rbs of Zymomonas mobilis ethanologenic genes (Theberge et al 1992;Rajnish et al 2008) and β-1,4-endoglucanase of Bacillus pumilus strain (Lima et al 2005), respectively.…”

Section: Discussionsupporting

confidence: 81%

Identification and characterization of endoglucanases for fungicidal activity in Anabaena laxa (Cyanobacteria)

et al. 2010

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A cyanobacterial strain (Anabaena laxa RPAN8) exhibiting fungicidal activity and β-1,3 and 1,4 endoglucanase activities was selected for identifying the gene(s) involved. Functional analyses of the genomic library revealed that four clones (8, 64, 116, and 248) of RPAN8 exhibited fungicidal activity and induced structural deformities in the cell wall of the growing mycelia of Pythium aphanidermatum. Higher expression of fungicidal and β-1,4 endoglucanase activities, along with low expression of β-1,3 endoglucanase activity, was recorded in two E. coli clones (8 and 64). Clones 8 and 64 exhibited identical sequences while clones 116 and 248 were also similar. Bioinformatic analyses were undertaken only for the two nonidentical clones 8 and 116 which showed open reading frames (ORFs) of 348 (end 1) and 656 amino acid residues (end 2), respectively. The amino acid sequence analyses revealed that the end 1 encoding endoglucanases belonged to peptidase M20 family while end 2 showed significant similarities with several known genes. The putative promoters and ribosomal binding sites were identified and amino acid exchanges were observed in both end 1 and 2. The presence of signal peptides of 24 and 20 amino acid residues respectively revealed the secretory nature of these proteins.

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Section: Discussionsupporting

confidence: 81%

Identification and characterization of endoglucanases for fungicidal activity in Anabaena laxa (Cyanobacteria)

et al. 2010

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“…In addition, similar growth inhibition was also observed in four ampicillin−resistant (AR) genes of ZMO0103 , ZMO0893 , ZMO1094 and the ZMO1650 knockout strain ZM4∆ARs. The previous study reported that ZMO0103 was a β−lactamase gene, which contains a 55% amino acid sequence identity with class C β−lactamase genes [ 20 ]. A higher expression level [ 30 , 34 , 35 , 36 ] of ZMO0103 in ZM4 may be ascribed to the absence of the AmpR that can inhibit the expression of ZMO0103 .…”

Section: Discussionmentioning

confidence: 99%

“…Although Z. mobilis is tolerant to ampicillin, only one work reported that ZMO0103 probably is an ampicillin−resistant gene, which reported the results of a heterologous protein expression and an enzymatic kinetic analysis [ 20 ]. The genome sequence of Z. mobilis ZM4 was published, and the genome annotation was further improved [ 21 , 22 , 23 ].…”

Section: Introductionmentioning

confidence: 99%

Identification and Characterization of Genes Related to Ampicillin Antibiotic Resistance in Zymomonas mobilis

Geng

Huang

et al. 2022

Antibiotics

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Antibiotics can inhibit or kill microorganisms, while microorganisms have evolved antibiotic resistance strategies to survive antibiotics. Zymomonas mobilis is an ideal industrial microbial chassis and can tolerate multiple antibiotics. However, the mechanisms of antibiotic resistance and genes associated with antibiotic resistance have not been fully analyzed and characterized. In this study, we investigated genes associated with antibiotic resistance using bioinformatic approaches and examined genes associated with ampicillin resistance using CRISPR/Cas12a−based genome−editing technology. Six ampicillin−resistant genes (ZMO0103, ZMO0893, ZMO1094, ZMO1650, ZMO1866, and ZMO1967) were identified, and five mutant strains ZM4∆0103, ZM4∆0893, ZM4∆1094, ZM4∆1650, and ZM4∆1866 were constructed. Additionally, a four−gene mutant ZM4∆ARs was constructed by knocking out ZMO0103, ZMO0893, ZMO1094, and ZMO1650 continuously. Cell growth, morphology, and transformation efficiency of mutant strains were examined. Our results show that the cell growth of ZM4∆0103 and ZM4∆ARs was significantly inhibited with 150 μg/mL ampicillin, and cells changed to a long filament shape from a short rod shape. Moreover, the transformation efficiencies of ZM4∆0103 and ZM4∆ARs were decreased. Our results indicate that ZMO0103 is the key to ampicillin resistance in Z. mobilis, and other ampicillin−resistant genes may have a synergetic effect with it. In summary, this study identified and characterized genes related to ampicillin resistance in Z. mobilis and laid a foundation for further study of other antibiotic resistance mechanisms.

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“…We have earlier reported that Z. mobilis showed high level of resistance (in clinical standards) to β-lactam antibiotics, such as cephalexin, cephalosporin C, cephalothin, benzylpenicillin and penicillin V (Rajnish et al 2011). The cloning and expression of a β-lactamase gene from Z. mobilis B406 in Escherichia coli JM 109 has been reported (Mukundan et al 1994).…”

Section: Introductionmentioning

confidence: 99%

“…The cloning and expression of a β-lactamase gene from Z. mobilis B406 in Escherichia coli JM 109 has been reported (Mukundan et al 1994). We have cloned, expressed, purified and characterized one of the β-lactamase (YP 161838) of Z. mobilis, in which the β-lactamase clone showed high level of resistance to ampicillin, benzylpenicillin, cephalexin and penicillin V (Rajnish et al 2011). We have also reported the homology based structural study of the β-lactam resistance determinants in Z. mobilis (Sheik Asraf et al 2012).…”

Section: Introductionmentioning

confidence: 99%

Computational and functional analysis of β-lactam resistance in Zymomonas mobilis

et al. 2013

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Zymomonas mobilis, a Gram-negative ethanologenic non-pathogenic bacterium, is reported to exhibit resistance to high concentrations of β-lactam antibiotics. In the present study, Z. mobilis was found to be resistant to I-IV generations of cephalosporins and carbapenems, i.e. narrow, broad and extended spectrum β-lactam antibiotics. We have analysed the genome of Z. mobilis (GenBank accession No.: NC 006526) harbouring multiple genes coding for β-lactamases (BLA), β-lactamase domain containing proteins (BDP) and penicillin binding proteins (PBP). The conserved domain database analysis of BDPs predicted them to be members of metallo β-lactamase superfamily. Further, class C specific multidomain AmpC (β-lactamase C) was found in the three β-lactamases. The β-lactam resistance determinants motifs, HXHXD, KXG, SXXK, SXN, and YXN are present in the BLAs, BDPs and PBPs of Z. mobilis. The predicted theoretical pI and aliphatic index values suggested their stability. One of the PBPs, PBP2, was predicted to share functional association with rod shape determining proteins (GenBank accession Nos. YP 162095 and YP 162091). Homology modelling of three dimensional structures of the β-lactam resistance determinants and further docking studies with penicillin and other β-lactam antibiotics indicated their substrate-specificity. Semi-quantitative PCR analysis indicated that the expression of all BLAs and one BDP are induced by penicillin. Disk diffusion assay, SDS-PAGE and zymogram analysis confirms the substrate specificity of the β-lactam resistance determinants. This study gives a broader picture of the β-lactam resistance determinants of a non-pathogenic ethanologenic Z. mobilis bacterium that could have implications in laboratories since it is routinely used in many research laboratories in the world for ethanol, fructooligosaccharides, levan production and has also been reported to be present in wine and beer as a spoilage organism.

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Functional characterization of a putative β-lactamase gene in the genome of Zymomonas mobilis

Cited by 4 publications

References 18 publications

Identification and characterization of endoglucanases for fungicidal activity in Anabaena laxa (Cyanobacteria)

Identification and characterization of endoglucanases for fungicidal activity in Anabaena laxa (Cyanobacteria)

Identification and Characterization of Genes Related to Ampicillin Antibiotic Resistance in Zymomonas mobilis

Computational and functional analysis of β-lactam resistance in Zymomonas mobilis

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