Exploration of a novel protease from Phytolacca dodecandra (Endod) berries for potential application in detergent formulation

Abstract

Conventional detergent formulations contain either petroleum-derived or synthetic chemicals. Hence, the replacement of these chemicals with natural products such as enzymes is considered a green technology. Among these enzymes, protease is renowned in detergent formulations. Therefore, the objective of this study was to explore and characterize a new protease from Phytolacca dodecandra, because the plant is traditionally known for detergency purposes. Enzyme extraction was performed with Glycine/NaOH buffer, pH = 10.0. Gelatin hydrolysis and agar well diffusion tests were implemented as qualitative assays, whereas the enzyme assay technique was applied for the quantitative investigation of the new protease from plant extracts. The crude enzyme was partially characterized concerning temperature and pH tolerance, washing performance, detergent compatibility, and stability. The finding of this study revealed that Phytolacca dodecandra contains protease and is characterized by 1418.84 U/mL of enzyme activity, where 1 unit (U) of enzyme activity refers to the amount of enzyme required to liberate 1 μg of tyrosine/mL/min under the defined assay conditions. The enzyme showed maximum activity at pH 10.5 and 45°C. The enzyme was stable and maintained 99.52% activity at 40°C and 91.73% at pH 10.0 after 2 h pre-incubation. The enzyme showed excellent wash performance and could potentially remove blood stains completely within 3 min. After 2 h of pre-incubation the enzyme maintained 69.06% and 54.88% of its activity in the presence of Azzy and Sunlight, respectively. The enzyme maintained 71.99% of its residual activity after 60 days of storage. If further studies are conducted, all these properties of the newly investigated protease can make the enzyme applicable in detergent formulations.