Characterization of recombinant glutelin type-B 5-like protein from proso millet

Research output: Contribution to conferencePaper


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.

Original languageEnglish
StatePublished - 2020
Event2020 ASABE Annual International Meeting - Virtual, Online
Duration: Jul 13 2020Jul 15 2020


Conference2020 ASABE Annual International Meeting
CityVirtual, Online

Bibliographical note

Funding 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.

Publisher Copyright:
© ASABE 2020 Annual International Meeting.


  • Proso millet
  • Protein purification
  • Recombinant DNA

ASJC Scopus subject areas

  • Agronomy and Crop Science
  • Bioengineering


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