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. 2013 Nov 28:2013:942590.
doi: 10.1155/2013/942590. eCollection 2013.

Production of Alkaline Protease by Solvent-Tolerant Alkaliphilic Bacillus circulans MTCC 7942 Isolated from Hydrocarbon Contaminated Habitat: Process Parameters Optimization

Ulhas Patil  1 , Ambalal Chaudhari  2
Affiliations

Production of Alkaline Protease by Solvent-Tolerant Alkaliphilic Bacillus circulans MTCC 7942 Isolated from Hydrocarbon Contaminated Habitat: Process Parameters Optimization

Ulhas Patil et al. ISRN Biochem. .

Abstract

In the present investigation, a newly isolated organic solvent-tolerant and alkaliphilic bacterial strain was reported from a hydrocarbon (gasoline and diesel) contaminated soil collected from the petrol station, Shirpur (India). The strain was identified as Bacillus circulans MTCC 7942, based on phenotype, biochemical, and phylogenetic analysis of 16S rRNA gene sequence. The capability of Bacillus circulans to secrete an extracellular, thermostable, alkaline protease and grow in the presence of organic solvents was explored. Bacillus circulans produced maximum alkaline protease (412 U/mL) in optimized medium (g/L): soybean meal, 15; starch, 10; KH2PO4, 1; MgSO4·7H2O, 0.05; CaCl2, 1; Na2CO3, 8; pH 10.0 at 37°C and 100 rpm. The competence of strain to grow in various organic solvents-n-octane, dodecane, n-decane, N,N-dimethylformamide, n-hexane, and dimethyl sulfoxide, establishes its potential as solvent-stable protease source for the possible applications in nonaqueous reactions and fine chemical synthesis.

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Figures

Figure 1
Figure 1
Phylogenetic tree of strain UB2 showing relatedness with Bacillus sp.
Figure 2
Figure 2
Dry cell weight of strain UB2 cultured in 100 mL of the nutrient liquid medium and 30 mL of organic solvent for 72 h.
Figure 3
Figure 3
Effect of ingredients of production medium on the growth and optimum protease production by B. circulans MTCC 7942 (a) soybean meal; (b) starch; and (c) glucose.
Figure 4
Figure 4
Effect of ingredients of production medium on the growth and optimum protease production by B. circulans MTCC 7942 (a) KH2PO4, (b) Na2CO3, and (c) CaCl2.
Figure 5
Figure 5
Effect of physical parameters on the growth and optimum protease production by B. circulans MTCC 7942 (a) temperature; (b) pH; (c) agitation; and (d) inoculum.
Figure 6
Figure 6
Profile of growth and protease production of B. circulans MTCC 7942 as a function of time.
See this image and copyright information in PMC

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