Expression, Purification, and Biophysical Characterization of Klebsiella Pneumoniae Nicotinate Nucleotide Adenylyltransferase

Autor: Tasvi Daya, Olamide Jeje, Reabetswe Maake, Chinyere Aloke, Thandeka Khoza, Ikechukwu Achilonu
Rok vydání: 2022
Předmět:
Zdroj: The Protein Journal. 41:141-156
ISSN: 1875-8355
1572-3887
DOI: 10.1007/s10930-021-10037-2
Popis: Patients in health-care settings develop nosocomial infections due to prolonged hospital stay. The Gram negative Klebsiella pneumoniae (K. pneumoniae), is a bacterial pathogen responsible for most nosocomial infections and are resistant to most current antibiotics. Hence, there is need for identification and validation of potential protein targets for design of new generation antibiotics. One of such targets is nicotinate nucleotide adenylyltransferase, an enzyme responsible for redox metabolism. This study focuses on novel expression, purification, and biophysical characterization of NNAT from K. pneumoniae. KpNNAT was over-expressed in T7 express™ Escherichia coli using the pGEX-4 T-1 expressions system and purified to 98% homogeneity (~ 20 mg KpNNAT/g of the wet cell) using a combination of glutathione-agarose and immobilized Ni
Databáze: OpenAIRE
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