title: Expression and purification of soluble bio-active rice plant catalase-A from recombinant Escherichia coli.
abstract: Catalase in plants is a heme-coordinated tetrameric protein that primarily disproportionates hydrogen peroxide into water and oxygen. It plays an important role in maintaining cellular concentration of hydrogen peroxide to a level, necessary for all aspects of normal plant growth and development. Except for its recombinant expression in transgenic plants and insect cell line, the protein is yet to be synthesized in its bio-active form in prokaryotic expression system. Attempts made in past for recombinant expression of plant catalase in Escherichia coli consistently resulted in formation of insoluble and inactive aggregates of inclusion body. Here we have shown the specific requirement of a thioredoxin fusion partner, the involvement of trigger factor protein and the low temperature treatment during induction period for synthesis of completely solubilized rice plant catalase-A in recombinant E. coli. Furthermore, the bacteria required the supplementation of δ-aminolevulinic acid to produce bio-active recombinant rice catalase-A. The molecular and biochemical properties of the purified recombinant protein showed the characteristic features of a typical mono-functional plant catalase. These results attest to the usefulness of the present protocol for production of plant catalase using E. coli as heterologous expression system.
authors: Gene Function and Regulation, Institute of Life Sciences, Nalco Square, Chandrasekharpur, Bhubaneswar 751023, Orissa, India.
authors: Mamata
authors: M
authors: Ray
authors: Panchanand
authors: P
authors: Mishra
authors: Priyanka
authors: P
authors: Das
authors: Surendra Chandra
authors: SC
authors: Sabat
mesh_terms: D002374:Catalase / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics; D006418:Heme / Q000378:metabolism; D018832:Molecular Chaperones; D012275:Oryza / Q000201:enzymology* / Q000235:genetics; D010940:Plant Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D012995:Solubility; D013879:Thioredoxins / Q000096:biosynthesis / Q000235:genetics
chemical_list: D018832:Molecular Chaperones; D010940:Plant Proteins; D011993:Recombinant Fusion Proteins; D006418:Heme; D013879:Thioredoxins; D002374:Catalase