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An improved method for extraction of sorghum polymeric protein complexes |
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| Institution: | 1. Institute for Food, Nutrition and Well-being, Department of Food Science, University of Pretoria, Private Bag X20, Hatfield 0028, South Africa;2. Institute of Food, Nutrition and Human Health, Massey University, Private Bag 11222, Palmerton North 4442, New Zealand;3. Department of Human Nutrition, University of Pretoria, P.O. Box 667, Pretoria 0001, South Africa;1. Center for Grain and Animal Health Research, USDA-ARS, Manhattan, KS, United States;2. School of Food Science, University of Idaho, Moscow, ID, United States |
| Abstract: | A method for fractionating sorghum proteins using extraction solvents and techniques designed to obtain polymeric protein structures (especially disulfide linked) was developed. Extraction and separation conditions were optimized in terms of completeness of protein extraction, sample stability, and analytical resolution. After pre-extraction of albumins and globulins, a 3-step sequential procedure involving no reducing agents was applied to ground whole sorghum flour. The three fractions obtained represented proportionally different protein polymer contents and molecular weight distribution as evidenced by comparative size exclusion chromatography. Protein composition also varied among the extracts with differences in kafirin composition and non-kafirin proteins detected in the fractions by RP-HPLC and SDS-PAGE analysis. The ability to quantify and further characterize sorghum polymeric protein complexes will be useful for additional studies linking protein structures with functionality and digestibility and variations for these properties within diverse sorghum germplasm. |
| Keywords: | Sorghum Protein Protein complexes Protein extraction Size exclusion chromatography Cereal Grain PMSF"} {"#name":"keyword" "$":{"id":"kwrd0050"} "$$":[{"#name":"text" "_":"Phenylmethylsulfonylfluoride PIC"} {"#name":"keyword" "$":{"id":"kwrd0060"} "$$":[{"#name":"text" "_":"Protease inhibitor cocktail SEC"} {"#name":"keyword" "$":{"id":"kwrd0070"} "$$":[{"#name":"text" "_":"Size-exclusion chromatography F1"} {"#name":"keyword" "$":{"id":"kwrd0080"} "$$":[{"#name":"text" "_":"Fraction 1 F2"} {"#name":"keyword" "$":{"id":"kwrd0090"} "$$":[{"#name":"text" "_":"Fraction 2 F3"} {"#name":"keyword" "$":{"id":"kwrd0100"} "$$":[{"#name":"text" "_":"Fraction 3 SDS"} {"#name":"keyword" "$":{"id":"kwrd0110"} "$$":[{"#name":"text" "_":"Sodium dodecyl sulfate RP-HPLC"} {"#name":"keyword" "$":{"id":"kwrd0120"} "$$":[{"#name":"text" "_":"Reversed phase – high performance liquid chromatography SDS-PAGE"} {"#name":"keyword" "$":{"id":"kwrd0130"} "$$":[{"#name":"text" "_":"Sodium dodecyl sulfate polyacrylamide gel electrophoresis |
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