Statistical media optimization for the production of clinical uricase from Bacillus subtilis strain SP6

Abstract

In this study, a potent uricase producing organism was isolated by a thorough screening and identified as Bacillus subtilis strain SP6 by using 16s rDNA sequencing. Response surface methodological optimization was employed for the enhanced production of uricase from newly isolated Bacillus subtilis strain SP6. In media optimization studies, Plackett Burman (PB) design was used for the selection of the critical media components; which were further optimized using central composite design (CCD). Lactose, soya peptone, uric acid and FeSO 4 .7H 2 O were found to be the critical factors influencing the enzyme production. Optimum uricase production with these factors was deduced using central composite design. Significant level of the factors were 12.2 g/L of lactose, 12.79 g/L of soya peptone, 2.55 g/L of uric acid and 0.00325 g/L FeSO 4 .7H 2 O. Use of statistical optimization upsurges uricase yield from 1.2 U/ml to 15.87 U/ml enhancing the overall production by 13.23 fold; which confirms that the model is effective for process optimization.

Optimization of industrial (3000 L) production of Bacillus subtilis CW-S and its novel application for minituber and industrial-grade potato cultivation

“…PBD and RSM have effectively optimized some bioprocesses 18 , 19 . In recent times, different researchers have published statistically optimized process designs of B. subtilis for the production of various enzymes and metabolites 20 , 21 . However, industrial production data on Bacillus subtilis as a plant probiotic is still limited.…”

Optimized UV-Spectrophotometric Assay to Screen Bacterial Uricase Activity Using Whole Cell Suspension

“…In few lactobacilli strains, uricase activity was suggested to be present both intracellularly and extracellularly ( Handayani et al, 2018 ), thus prompting further investigation of uricase in taxa associated with probiotic strains. The determination of bacterial uricase activity can be assessed either qualitatively, via the observation of UA consumption on UA-containing solid media ( Shaaban et al, 2015 ; Pustake et al, 2019 ), or quantitatively, via the measurement of H 2 O 2 formation using fluorescent dyes ( Fraisse et al, 2002 ) or the determination of UA degradation using spectrophotometry ( Koyama et al, 1996 ; Fraisse et al, 2002 ; Huang and Wu, 2004 ) or liquid chromatography ( Safranow et al, 2000 ). Because of their simplicity and compatibility with microplate systems allowing parallel analysis of multiple samples, spectrophotometric and fluorescent techniques appear promising to develop a high-throughput screening assay for bacterial uricase activity.…”

Isolation and Characterization of Uricase Produced from Chicken Liver

“…Several studies have carried out the isolation of the uricase in plants, microorganisms, and animals. Isolation of uricase from green bean leaves [ [4]], Pseudomonas aeruginosa [ [5]], Streptomyces exfoliates UR10 from agricultural waste [ [6]], Bacillus subtilis strain SP6 [ [7]], Bacillus cereus SKIII [ [8]], Aspergillus welwitschiae strain 1-4 [ [9]]. Isolation of the uricase from the liver of rainbow fish, mackerel, trout, catfish, shark and tilapia [ [10]], Indonesian coelacanth [ [11]], camel liver [ [12]], and bovine kidney [ [13]].…”