A set of simple methods for detection and extraction of laminarinase

Koteshwara, Ananthamurthy and Philip, Nancy V and Mathew, Jesil A and Chandrashekhar, Raghu H and Subrahmanyam, VM (2021) A set of simple methods for detection and extraction of laminarinase. Scientific Reports, 11 (1). pp. 1-9. ISSN 2045-2322

[img] PDF
14137.pdf - Published Version
Restricted to Registered users only

Download (1MB) | Request a copy

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 purifcation. 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 fuorescent 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

Item Type: Article
Subjects: Pharmacy > MCOPS Manipal > Pharmaceutical Biotechnology
Depositing User: KMC Library
Date Deposited: 28 Jan 2022 03:50
Last Modified: 28 Jan 2022 03:50
URI: http://eprints.manipal.edu/id/eprint/158196

Actions (login required)

View Item View Item