title: A novel alkaline keratinase from Bacillus subtilis DP1 with potential utility in cosmetic formulation.
abstract: The Bacillus subtilis DP1 was isolated from poultry farm soil at Anand district, India. The highest enzyme production (379.65U/ml) was obtained at pH 10.0, a temperature of 37°C and a growth period of 72h. The extracellular keratinase was purified by gel filtration chromatography with 27.98 purification fold. Purity was also confirmed by High-Performance Liquid Chromatography (HPLC) analysis, where a major peak having retention time of 2.5min was obtained on C18 column using photo diode array detector. Purified keratinase was stable in a broad range of pH (8-12) and temperature (20-50°C) with optimum at pH 10.0 and 37°C. The metallic ions, Ca(2+) and Mg(2+) enhance keratinase activity. Secondary structure from Circular Dichroism (CD) spectra implies that purified keratinase is largely β-pleated sheet rich protein. For preparation of dehairing cream formulation, compatibility studies of excipients were carried out. Fourier transform infrared spectroscopy (FTIR) spectra of sodium stearate, calcium carbonate and sodium lauryl sulphate shows no reactivity of functional groups and hence mixture was compatible for formulation of keratinase dehairing cream. Prepared biological depilatory was able to remove hair more efficiently compared to marketed formulations.
authors.affiliation: Department of Pharmaceutical Sciences, Saurashtra Univeristy, Rajkot-360005, India. Electronic address: gv.sanghvi@gmail.com.
authors.forename: Gaurav
authors.initials: G
authors.lastname: Sanghvi
authors.affiliation: Department of Pharmaceutical Sciences, Saurashtra Univeristy, Rajkot-360005, India.
authors.forename: Harshit
authors.initials: H
authors.lastname: Patel
authors.affiliation: Department of Pharmaceutical Sciences, Saurashtra Univeristy, Rajkot-360005, India.
authors.forename: Devendra
authors.initials: D
authors.lastname: Vaishnav
authors.affiliation: Department of Pharmaceutical Sciences, Saurashtra Univeristy, Rajkot-360005, India.
authors.forename: Tejas
authors.initials: T
authors.lastname: Oza
authors.affiliation: Department of Biochemistry, Saurashtra University, Rajkot-360005, India.
authors.forename: Gaurav
authors.initials: G
authors.lastname: Dave
authors.affiliation: B. N. Patel Institute of Paramedical and Sciences, Bhalej Road, Anand, India.
authors.forename: Prashant
authors.initials: P
authors.lastname: Kunjadia
authors.affiliation: Department of Pharmaceutical Sciences, Saurashtra Univeristy, Rajkot-360005, India.
authors.forename: Navin
authors.initials: N
authors.lastname: Sheth
mesh_terms: D000818:Animals; D001412:Bacillus subtilis / Q000201:enzymology* / Q000378:metabolism; D002645:Chickens; D003358:Cosmetics / Q000737:chemistry*; D004339:Drug Compounding; D006204:Hair Removal; D006863:Hydrogen-Ion Concentration; D010447:Peptide Hydrolases / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification; D017433:Protein Structure, Secondary; D013696:Temperature
chemical_list: D003358:Cosmetics; D010447:Peptide Hydrolases; C035855:keratinase
keywords: Alkaline keratinase; Bacillus subtilis DP1; CD spectra; Dehairing cream