title: Purification and Characterization of a Protease Produced by a Planomicrobium sp. L-2 from Gut of Octopus vulgaris.
abstract: Protease widely exists in the digestive tract of animals and humans, playing a very important role in protein digestion and absorption. In this study, a high protease-producing strain Planomicrobium sp. L-2 was isolated and identified from the digestive tract of Octopus variabilis. The strain was identified by physiological and biochemical experiments and 16S rDNA sequences analysis. A protease was obtained from the strain Planomicrobium sp. L-2 through ammonium sulfate precipitation, dialysis and enrichment, DEAE-Sephadex A50 anion-exchange chromatography, and Sephadex G-100 gel chromatography. The molecular weight and properties of the protease were characterized, including optimum temperature and pH, thermal stability, protease inhibitions and metal ions. According to our results, the protease from Planomicrobium sp. L-2 strain designated as F1-1 was obtained by three-step separation and purification from crude enzyme. The molecular weight of the protease was 61.4 kDa and its optimum temperature was 40°C. The protease F1-1 showed a broad pH profile for casein hydrolysis between 5.0~11.0. No residual activity was observed after incubation for 40 min at 60°C and 60 min at 50°C. F1-1 protease was inhibited by Mn(2+), Hg(2+), Pb(2+), Zn(2+), and Cu(2+) ions, as well as PMSF, indicating that the protease F1-1 was a serine protease. Additionally, research basis provided by this study could be considered for industrial application of octopus intestinal proteases.
authors.affiliation: Department of Food Science and Technology, College of Fisheries Sciences, Pukyong National University, Busan 608-737, Korea.
authors.forename: Qing
authors.initials: Q
authors.lastname: Liu
authors.affiliation: Department of Food Science and Technology, Qingdao Agricultural University, Qingdao 266109, China.
authors.forename: Shujing
authors.initials: S
authors.lastname: Sun
authors.affiliation: Department of Food Science and Technology, Qingdao Agricultural University, Qingdao 266109, China.
authors.forename: Meizi
authors.initials: M
authors.lastname: Piao
authors.affiliation: Department of Food Science and Technology, College of Fisheries Sciences, Pukyong National University, Busan 608-737, Korea.
authors.forename: Ji Young
authors.initials: JY
authors.lastname: Yang
keywords: enzymatic properties; identification; intestinal bacteria; octopus; purification