Proso millet glutelin protein fractions make up its second-largest protein mostly used in proso millet concentrates and isolates. In this experiment, we identified the gene of glutelin type-B 5-like protein isoform of the glutelin fraction with over 40% structural identity with the crystal structure of recombinant pro-11S globulin of pumpkin (2E9Q). We further cloned the gene of glutelin type-B 5-like protein with attached six continuous histidine codons into a pET 21d expression vector, overexpressed in rosetta Escherichia coli cells, purified the protein from inclusion bodies by unfolding with 8 M urea and refolding in 0.5 M Arginine with 20 mM CHES (2-(Cyclohexylamino)ethanesulfonic acid). Characterization of purified protein reveals above 90% pure protein based on an SDS-PAGE gel analysis with a band 22.1 kDa. Additionally, the protein revealed a monodispersed particle size distribution with a broad range of particle size (6.50 - 43.82 nm) and a higher-order aggregation with molecular weight range 60-70 kDa.
|State||Published - 2020|
|Event||2020 ASABE Annual International Meeting - Virtual, Online|
Duration: Jul 13 2020 → Jul 15 2020
|Conference||2020 ASABE Annual International Meeting|
|Period||7/13/20 → 7/15/20|
Bibliographical noteFunding Information:
This work was supported by the Kentucky Agricultural Experiment Station (KAES), and the National Institute of Food and Agriculture (NIFA), U.S. Department of Agriculture, Hatch-Multistate project #: 1007893. Additionally, authors would like to thank Catherine Chaton, Zamakhaeva Svetlana, Williamson Zachary, and Shakhashiro Muna for their contributions to this project.
© ASABE 2020 Annual International Meeting.
- Proso millet
- Protein purification
- Recombinant DNA
ASJC Scopus subject areas
- Agronomy and Crop Science