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Production and Utilization of Locally Characterized Recombinant β-Lactamase as Positive Control in the Antibiotic Susceptibility Testing

Production and Utilization of Locally Characterized Recombinant β-Lactamase as Positive Control in the Antibiotic Susceptibility Testing

Amjed Ali1, Muhammad Tayyab1,*, Sehrish Firyal1, Abu Saeed Hashmi2, Asif Nadeem3, Shagufta Saeed1, Ali Raza Awan1 and Muhammad Wasim1

1Institute of Biochemistry and Biotechnology, University of Veterinary and Animal Sciences, Abdul Qadir Jillani Road, Lahore
2Riphah International University, Raiwind Road Campus, Lahore 
3Department of Biotechnology, Virtual University, Lahore

*     Corresponding author: tayyab_pakistan@yahoo.com; muhammad.tayyab@uvas.edu.pk

ABSTRACT

Antibiotic susceptibility testing is the most suitable strategy for antibiotic resistance detection in bacteria. This project was an initial step for the production of β-lactamase for clinical diagnostics in Pakistan. The current work deals with the optimization of conditions for higher level production of recombinant β-lactamase from Bacillus subtilis R5. Supplementation of LB medium with various carbon sources including wheat bran, rice bran and molasses could increase the enzyme production from 25 to 43.7, 44.3 and 49 µmole/min whereas the nitrogen sources including tryptone, peptone and yeast extract could enhance the enzyme production from 25 to 40.21, 41 and 43.76 µmole/min, respectively. The highest β-lactamase production was recorded when the LB medium was supplemented with 4% wheat bran and 2% yeast extract. The locally produced recombinant β-lactamase exhibited strong potential regarding the hydrolysis of β-lactam ring containing antibiotics and showed comparable results to β-lactamase from Hi-media kit available in the market. The β-lactamase from current study was found suitable for its use as positive control in antibiotic susceptibility testing and this enzyme will be utilized for the development of antibiotic susceptibility testing kit at domestic level in near future.
 

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Pakistan Journal of Zoology

December

Vol. 53, Iss. 6, Pages 2001-2521

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