title: Tobacco phytaspase: Successful expression in a heterologous system.
abstract: Phytaspase, a plant serine protease, has been demonstrated to play an important role in the programmed cell death of various plants. Phytaspase is synthesized as an inactive proenzyme containing an N-terminal signal peptide followed by a pro-domain and a mature protease catalytic domain. Pre-prophytaspase autocatalytically processes itself into a pro-domain and an active mature phytaspase enzyme. We have recently demonstrated the successful expression of mature phytaspase from tobacco in a bacterial system. Herein, we focus on the expression of pre-prophytaspase as a GST-tag fusion and on its purification by affinity chromatography.
authors.affiliation: a Department of Biosciences & Bioengineering , Guwahati-39, Assam , India.
authors.forename: Sharmila
authors.initials: S
authors.lastname: Narayanan
authors.affiliation: b Centre for Nanotechnology, Indian Institute of Technology Guwahati , Guwahati-39, Assam , India.
authors.forename: Pallab
authors.initials: P
authors.lastname: Sanpui
authors.affiliation: a Department of Biosciences & Bioengineering , Guwahati-39, Assam , India.
authors.forename: Lingaraj
authors.initials: L
authors.lastname: Sahoo
authors.affiliation: a Department of Biosciences & Bioengineering , Guwahati-39, Assam , India.|b Centre for Nanotechnology, Indian Institute of Technology Guwahati , Guwahati-39, Assam , India.
authors.forename: Siddhartha Sankar
authors.initials: SS
authors.lastname: Ghosh
mesh_terms: D003001:Cloning, Molecular / Q000379:methods; D004926:Escherichia coli / Q000502:physiology*; D010833:Phytic Acid / Q000096:biosynthesis*; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D057057:Serine Proteases / Q000096:biosynthesis* / Q000235:genetics* / Q000302:isolation & purification; D014026:Nicotiana / Q000201:enzymology* / Q000235:genetics*
chemical_list: D011994:Recombinant Proteins; D010833:Phytic Acid; D057057:Serine Proteases
keywords: affinity chromatography; caspases; heterologous expression; phytaspase; programmed cell death; serine protease; subtilases