|Park, S - KANSAS STATE UNIV|
Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: June 19, 2002
Publication Date: October 13, 2002
Citation: Bean, S., Park, S. 2002. Optimized methods for extraction and sds-page analysis of sorghum proteins. Abstract No. 288 in: 2001 AACC Annual Meeting Program Book. p.132. Meeting Abstract. Interpretive Summary: Abstract for AACC Annual Meeting to be held October 13-17, 2002 in Montreal, Canada.
Technical Abstract: Gradient gels are commonly used for analyzing sorghum proteins and, while effective, are difficult and time consuming to cast. Therefore this research evaluated the use of precast gels for the separation of sorghum proteins and optimization of protein extraction with the goal of decreasing the total time necessary for analysis. The precast gels used in this project utilized novel buffer systems at neutral pH which had improved stability and increased resolution relative to traditional gels. Two buffers were used (MOPS and MES) with 12% and 4-12% gels. Good separations were achieved using 4-12% gels with either buffer. This simplified the analysis of sorghum proteins by SDS-PAGE and produced high resolution separations. To optimize the extraction of sorghum proteins, several variables were examined: sample to solvent ratio, detergent type and concentration, reducing agent type and concentration, extraction time, and buffer pH and concentration. Samples were quantified and characterized by RP-HPLC, SDS-PAGE and FZCE. These studies revealed that pH, detergent type, reducing agent type and sample to solvent ratio all had significant effect on levels of protein extracted. Increasing SDS concentration (2%), and solvent to flour ratio (20:1) with multiple 5-10 min extracts reduced extraction time by ~50-75% while still extracting ~94% of total protein relative to the control methodology.