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L6170

Sigma-Aldrich

β-Lactamase

recombinant, expressed in E. coli

Synonym(s):

Carbapenemase, Cephalosprinase

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About This Item

CAS Number:
Enzyme Commission number:
EC Number:
UNSPSC Code:
12352204
NACRES:
NA.54

recombinant

expressed in E. coli

Quality Level

form

powder

specific activity

≥20 U/mg (with cephalosporin C)
≥400 U/mg (with benzylpenicilin)

storage temp.

2-8°C

General description

β-Lactamase produced by bacteria is closely related to the penicillin binding proteins. β-Lactamase hydrolyzes β-lactum antibiotics and is the prime cause of resistance development by bacteria. There are four subclasses of β-Lactamases. Classes A, C and D form an acyl-enzyme via active site serine residue. Class B β-lactamases are metalloenzymes with zinc ion at their active site for β-lactam hydrolysis. Mutations in the β-lactamases has resulted in the generation of extended-spectrum β-lactamases (ESBLs). As close to 900 types of β-Lactamases are produced by microbes.

Application

β--lactamase is used to inactivate β-lactam antibiotics by breaking open the β-lactam ring. β--lactamase is used to study antibiotic resistance and resistance suppression. Product L6170 is recombinantly produced based on the sequence of the enzyme from Pseudomonas aeruginosa, and is expressed in E. coli.
β-Lactamase has been used in coumarin-cephalosporin-fluorescein-acetoxymethyl (CCF4)- β-Lactamase assay in dendritic cells and in the hydrolysis of meropenem (H-Mer) to generate 18O-labeled H-Mer.

Biochem/physiol Actions

β--lactamase inactivates β-lactam antibiotics by breaking open the β-lactam ring. Typical analysis for other substrates: ceftriaxon 2-4 un/mg; cefazolin 2-4 un/mg; ceftazidime 1.5-3 un/mg; cefoxitin 0.35-0.7 un/mg; cefepime 1.2-2.4 un/mg; cefuroxime 1.5-3 un/mg; cefotaxime 1-2 un/mg, oxacillin 4-8 un/mg; imipenem 1.2-2.4 un/mg; and meropenem 3-6 un/mg
Typical analysis for other substrates: ceftriaxon 2-4 un/mg; cefazolin 2-4 un/mg; ceftazidime 1.5-3 un/mg; cefoxitin 0.35-0.7 un/mg; cefepime 1.2-2.4 un/mg; cefuroxime 1.5-3 un/mg; cefotaxime 1-2 un/mg, oxacillin 4-8 un/mg; imipenem 1.2-2.4 un/mg; and meropenem 3-6 un/mg

Unit Definition

One unit will hydrolyze 1.0 μmole substrate per min (βI: benzylpenicillin; βII: cephalosporin C) in 50 mM phosphate buffer, pH 7.0 containing 10 μM zinc chloride at 25 ºC.

Physical form

Lyophilized powder containing sodium chloride and potassium phosphate.

Storage Class Code

11 - Combustible Solids

WGK

WGK 3

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable


Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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Updated functional classification of beta-lactamases
Bush K and Jacoby GA
Antimicrobial Agents and Chemotherapy, 54(3), 969-976 (2010)
Determining carbapenemase activity with 18O labeling and targeted mass spectrometry
Wang M, et al.
Analytical Chemistry, 85(22), 11014-11019 (2013)
VAMP8-mediated NOX2 recruitment to endosomes is necessary for antigen release
Dingjan I, et al.
European Journal of Cell Biology, 96(7), 705-714 (2017)
Antibiotic resistance and extended spectrum beta-lactamases: Types, epidemiology and treatment
Shaikh S, et al.
Saudi Journal of Biological Sciences, 22(1), 90-101 (2015)
Arnold Louie et al.
Antimicrobial agents and chemotherapy, 56(1), 258-270 (2011-10-26)
New broad-spectrum β-lactamases such as KPC enzymes and CTX-M-15 enzymes threaten to markedly reduce the utility of our armamentarium of β-lactam agents, even our most potent drugs, such as carbapenems. NXL104 is a broad-spectrum non-β-lactam β-lactamase inhibitor. In this evaluation

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