Bioprocessing of “Hair Waste” by Paecilomyces lilacinus as a Source of a Bleach-Stable, Alkaline, and Thermostable Keratinase with Potential Application as a Laundry Detergent Additive: Characterization and Wash Performance Analysis
Paecilomyces lilacinus (Thom) Samson LPS 876, a locally isolated fungal strain, was grown on minimal mineral medium containing “hair waste,” a residue from the hair-saving unhairing process, and produced a protease with keratinolytic activity. This enzyme was biochemically characterized. The optimum reaction conditions, determined with a response surface methodology, were 60°C and pH 6.0. It was remarkably stable in a wide range of pHs and temperatures. Addition of Ca2+, Mg2+, or sorbitol was found to be effective in increasing thermal stability of the protease. PMSF and Hg2+ inhibited the proteolytic activity indicating the presence of a thiol-dependent serine protease. It showed high stability toward surfactants, bleaching agents, and solvents. It was also compatible with commercial detergents (7？mg/mL) such as Ariel, Skip, Drive, and Ace, retaining more than 70% of its proteolytic activity in all detergents after 1？h of incubation at 40°C. Wash performance analysis revealed that this protease could effectively remove blood stains. From these properties, this enzyme may be considered as a potential candidate for future use in biotechnological processes, as well as in the formulation of laundry detergents. 1. Introduction Microbial proteases are the most widely exploited industrial enzymes with major application in detergent formulations [1, 2]. These enzymes are being widely used in detergent industry since their introduction in 1914 as detergent additive. Over the past 30 years, the importance of proteases in detergents has changed from being the minor additives to being the key ingredients. The main areas where use of proteases has expanded are household laundry, automatic dishwashers, and industrial and institutional cleaning. In laundry detergents, protein stains such as grass, blood, food, and human swear, are removed through proteolysis. The performance of proteases is influenced by several factors such as pH of detergent, ionic strength, wash temperature, detergent composition, bleach systems, and mechanical handling. Thus, the key challenge for the use of enzymes in detergents is their stability. Various attempts have been made to enhance stability of alkaline proteases by site-directed mutagenesis [3] and protein engineering. “Subtilisin Carlsberg” has been protein engineered to obtain a bleach-stable, alkaline protease by molecular modification [4], but still, there is always a need for newer thermostable alkaline proteases which can withstand bleaching agents present in detergent. Among these different proteases, keratinases constitute a
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