Purification of Recombinant Xylosyltransferase XXT2 involved in Xyloglucan Biosynthesis
Date
Authors
Major Professor
Advisor
Committee Member
Journal Title
Journal ISSN
Volume Title
Publisher
Authors
Research Projects
Organizational Units
The Department of Biochemistry, Biophysics, and Molecular Biology was founded to give students an understanding of life principles through the understanding of chemical and physical principles. Among these principles are frontiers of biotechnology such as metabolic networking, the structure of hormones and proteins, genomics, and the like.
History
The Department of Biochemistry and Biophysics was founded in 1959, and was administered by the College of Sciences and Humanities (later, College of Liberal Arts & Sciences). In 1979 it became co-administered by the Department of Agriculture (later, College of Agriculture and Life Sciences). In 1998 its name changed to the Department of Biochemistry, Biophysics, and Molecular Biology.
Dates of Existence
1959–present
Historical Names
- Department of Biochemistry and Biophysics (1959–1998)
Related Units
- College of Agriculture and Life Sciences (parent college)
- College of Liberal Arts and Sciences (parent college)
Journal Issue
Is Version Of
Versions
Series
The Symposium provides undergraduates from all academic disciplines with an opportunity to share their research with the university community and other guests through conference-style oral presentations. The Symposium represents part of a larger effort of Iowa State University to enhance, support, and celebrate undergraduate research activity.
Though coordinated by the University Honors Program, all undergraduate students are eligible and encouraged to participate in the Symposium. Undergraduates conducting research but not yet ready to present their work are encouraged to attend the Symposium to learn about the presentation process and students not currently involved in research are encouraged to attend the Symposium to learn about the broad range of undergraduate research activities that are taking place at ISU.
The first Symposium was held in April 2007. The 39 students who presented research and their mentors collectively represented all of ISU's Colleges: Agriculture and Life Sciences, Business, Design, Engineering, Human Sciences, Liberal Arts and Sciences, Veterinary Medicine, and the Graduate College. The event has grown to regularly include more than 100 students presenting on topics that span the broad range of disciplines studied at ISU.
Department
Abstract
Plant cell walls are important in society as food, fuel, and fiber. However, many aspects of cell wall biosynthesis remain unknown. The polysaccharide xyloglucan plays an important role in the structure of the primary cell wall of dicotyledonous plants and is the most abundant hemicellulose in dicots. Xyloglucan biosynthesis involves at least seven different type II transmembrane glycosyl transferases in the Golgi. The goal of this research is to determine the conditions to successfully express and purify the soluble fully-folded recombinant XXT2, a critical xylosyltransferase involved in xyloglucan biosynthesis. Using E. Coli to express the protein with an attached tag, affinity and size-exclusion chromatography was used to purify the protein, then it was digested to remove the tag from the protein. To confirm the activity and the correct conformation of the enzyme, HPLC and MALDI-TOF were used. By expressing and purifying the folded, soluble proteins, protein-protein interactions will be investigated, the residues involved in these interactions will be determined, and also proteins can be prepared for structural characterization using X-ray crystallography. This research will help to understand the polysaccharide biosynthesis, and also assist in the plant cell wall modifications to improve biomass properties improvements for industrial applications.