Extraction and Purification of Laccases from Rice Stems

Chenna Swetha; P V Shivaprasad

doi:10.21769/BioProtoc.3208

Extraction and Purification of Laccases from Rice Stems

Bio Protoc. 2019 Apr 5;9(7):e3208. doi: 10.21769/BioProtoc.3208.

Authors

Chenna Swetha^{1

2}, P V Shivaprasad¹

Affiliations

¹ National Centre for Biological Sciences, Tata Institute of Fundamental Research, GKVK Campus, Bangalore 560065, India.
² SASTRA University, Thirumalaisamudram, Thanjavur, 613401, India.

Abstract

Laccases are found in cell walls of plants in very low amounts. This protocol provides an efficient method to purify laccases from rice stems. The method involves three steps: 1) Isolation of total protein from rice stems using buffers with high salt concentration to extract protein from cell walls; 2) Purification of laccases using concanavalin-A beads; and, 3) In-gel staining of laccases with 4-hydroxyindole. Concanavalin-A specifically binds to internal or non-reducing terminal α-D-mannosyl and α-D-glucosyl groups found in glycoproteins and glycolipids. Laccases being glycoproteins binds to concanavalin-A during purification process and eluted with mannose.

Keywords: Cell wall; Laccase; Lignin; Phenylpropanoid pathway; Plants; Rice.