The biotechnological potential of subtilisin‐like fibrinolytic enzyme from a newly isolated Lactobacillus plantarum KSK‐II in blood destaining and antimicrobials

Abstract

An antimicrobial oxidative- and SDS-stable fibrinolytic alkaline protease designated as KSK-II was produced by Lactobacillus plantarum KSK-II isolated from kishk, a traditional Egyptian food. Maximum enzyme productivity was obtained in medium containing 1% lactose and 0.5% soybean flour as carbon and nitrogen sources, respectively. Purification of enzyme increased its specific activity to 1,140-fold with a recovery of 33% and molecular weight of 43.6 kDa. Enzyme activity was totally lost in the presence of ethylenediaminetetraacetic acid and was restored after addition of Fe(2+) suggesting that KSK-II is a metalloprotease and Fe(2+) acts as cofactor. Enzyme hydrolyzed not only the natural proteins but also synthetic substrates, particularly Suc-Ala-Ala-Pro-Phe-pNA. KSK-II can hydrolyze the Lys-X easier than Arg-X; thus, it was considered as a subtilisin-family protease. Its apparent Km , Vmax , and Kcat were 0.41 mM, 6.4 µmol mg(-1) min(-1) , and 28.0 s(-1) , respectively. KSK-II is industrially important from the perspectives of its maximal activity at 50°C (stable up to 70°C), ability to function at alkaline pH (10.0), stability at broad pH ranges (7.5-12.0) in addition to its stability toward SDS, H2 O2 , organic solvents, and detergents. We emphasize for the first time the potential of fibrinolytic activity for alkaline proteases used in detergents especially in blood destaining.

Prospects of fibrinolytic proteases of bacteria from sea cucumber fermentation products as antithrombotic agent

“…In the last decades, many thrombolytic agents have been identified, studied and characterized from various sources such as earthworms, snakes, fungi, and bacteria. Bacteria have the potential to produce economically valuable enzymes with several advantages, including faster growth, scale of cell production will be more easily increased, production conditions do not depend on the season, short-time required, low cost, high activity, safe and not toxic and easily manipulated genetically [4,6,7]. Therefore, research about anti thrombolytic agents from new fibrinolytic protease producing bacteria must be done because more effective and efficient anti thrombolytic agents are still needed.…”

In vitro and in silico characterization of alkaline serine protease from Bacillus subtilis D9 recovered from Saudi Arabia

“…Bacterial isolate D9 was selected for further microbiological and biochemical investigation, as it produced the highest clearance zone around its growing colonies on casein agar. The qualitative screening of the alkaline protease (AKD9) of isolate D9 was carried out on the basal medium described by Kotb (2015) containing (g/L): Soybean powder (5), lactose sugar (10), K 2 HPO 4 (0.5), KH 2 PO 4 (1.5), CaCl 2 (2), and MgSO 4 .7H 2 O (0.5). The growth was allowed for 48 h at 40 °C in a 50 mL medium (pH 9.5).…”

Histological Studies on a Newly Isolated Bacillus subtilis D10 Protease in the Debridement of Burn Wound Eschars Using Mouse Model

“…The potential proteolytic enzyme from the local isolate number D10 was produced using the basal medium described by Kotb [ 19 ]. The organism was grown for 56 h at 40 °C in a 50 mL solution (pH 9.0) comprising of (g/L distilled water): soybean powder (5), lactose sugar (10), K 2 HPO 4 (0.5), KH 2 PO 4 (1.5), CaCl 2 (2), and MgSO 4 .7H 2 O (0.5).…”