The optimization of physiochemical parameters for alkaline protease production using Bacillus licheniformis NCIM 2042 were carried out by Plackett-Burman design and response surface methodology (RSM). The model was validated experimentally and the maximum protease production was found 315.28 U using optimum culture conditions. The protease was purified using ammonium sulphate (60%) precipitation technique. The HPLC analysis of dialyzed sample showed that the retention time is 1.84？min with 73.5% purity. This enzyme retained more than 92% of its initial activity after preincubation for 30？min at in the presence of 25%？v/v DMSO, methanol, ethanol, ACN, 2-propanol, benzene, toluene, and hexane. In addition, partially purified enzyme showed remarkable stability for 60？min at room temperature, in the presence of anionic detergent (Tween-80 and Triton X-100), surfactant (SDS), bleaching agent (sodium perborate and hydrogen peroxide), and anti-redeposition agents (Na2CMC, Na2CO3). Purified enzyme containing 10%？w/v PEG 4000 showed better thermal, surfactant, and local detergent stability. 1. Introduction The protease is ubiquitous in nature. It is found in all living organisms and required for cell growth and differentiation. Alkaline proteases are one of the most important groups of industrial enzymes. They are extensively used in leather, food, pharmaceutical, textile, organic chemical synthesis, wastewater treatment, and other industries [1]. Alkaline proteases hold a major share of the enzyme market with two-third share in detergent industry alone [2, 3]. Due to enhancement of such demand of proteases for specific properties, scientists are looking for newer sources of proteases. For effective use in industries, alkaline proteases need to be stable and active at high temperature and pH and in the presence of surfactants, oxidizing agents, and organic solvents [4–7]. Although there are many microbial sources available for protease production, only a few are considered as commercial producers [8]. Of these, species of Bacillus dominate in the industry [9]. Only large-scale production of alkaline protease can fulfill the demand and usefulness of the proteases in the industry. In industry, microbial protease production was carried out by fermentative process. It is necessary to improve the yield of protease without increasing the process cost through fermentative process. Rapid enzyme production can be achieved by manipulation of media composition and culture conditions. Thus, optimization of fermentation conditions is the most important step in the
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