A set of simple methods for detection and extraction of laminarinase

Abstract

A carefully designed ammonium sulfate precipitation will simplify extraction of proteins and is considered to be a gold standard among various precipitation methods. Therefore, optimization of ammonium sulfate precipitation can be an important functional step in protein purification. The presence of high amounts of ammonium sulphate precludes direct detection of many enzymatically active proteins including reducing sugar assays (e.g. Nelson-Somogyi, Reissig and 3,5-dinitrosalicylic acid methods) for assessing carbohydrases (e.g. laminarinase (β (1–3)-glucanohydrolase), cellulases and chitinases). In this study, a simple method was developed using laminarin infused agarose plate for the direct analysis of the ammonium sulphate precipitates from Streptomyces rimosus AFM-1. The developed method is simple and convenient that can give accurate results even in presence of ammonium sulfate in the crude precipitates. Laminarin is a translucent substrate requiring the use of a stain to visualize the zones of hydrolysis in a plate assay. A very low-cost and locally available fluorescent optical fabric brightener Tinopal CBS-X has been used as a stain to detect the zones of hydrolysis. We also report simple methods to prepare colloidal chitin and cell free supernatant in this manuscript.

Purification of bromelain enzyme from MD2 hybrid pineapple core by ultrafiltration and its antioxidative potential

“…Conversely, the proteolytic activity and specific activity of PPB are lower than that of crude even though the crude had higher protein content than PPB. The low proteolytic and specific activity of PPB was due to the presence of excess ammonium sulfate salt that hinders the enzyme's active site, causing less substrate binding to the active site (Junaidi et al, 2017;Koteshwara et al, 2021). The purification fold of PPB was decreased with a folding of 0.20, whereas UFB has a purification fold of 3.25.…”

No Solid Colloidal Carriers: Aspects Thermodynamic the Immobilization Chitinase and Laminarinase in Liposome

“…Laminarinases and chitinases are important enzymes responsible for the lysis of the fungal cell and sclerotial wall in fungi such as Trichoderma spp (Koteshwara et al, 2021). Different phospholipids, such as soya lecithin, which is made up of various triglycerides, phospholipids (phosphatidylinositol, phosphatidyl choline, phosphatidyl ethanol amine), and glycolipids, can be utilized to make liposomes (Jørgensen et al, 2004;Li et al, 2015).…”

Cold-Active Lipases and Esterases: A Review on Recombinant Overexpression and Other Essential Issues

“…In general, obtaining high-purity recombinant enzymes in their stable and active form is expensive, time-consuming, and complex. One-step purification using ammonium sulfate is usually term as partial purification; a well-designed ammonium sulfate precipitation is regarded as a gold standard among several purification strategies [ 96 ].…”