Single-step purification of native miraculin using immobilized metal-affinity chromatography.
Miraculin is a taste-modifying protein that can be isolated from miracle fruit ( Richadella dulcifica ), a shrub native to West Africa. It is able to turn a sour taste into a sweet taste. The commercial exploitation of this sweetness-modifying protein is underway, and a fast and efficient purification method to extract the protein is needed. We succeeded in purifying miraculin from miracle fruit in a single-step purification using immobilized metal-affinity chromatography (IMAC). The purified miraculin exhibited high purity (>95%) in reverse-phase high-performance liquid chromatography. We also demonstrated the necessity of its structure for binding to the nickel-IMAC column.
Duhita
Narendra
N
Graduate School of Life and Environmental Sciences, University of Tsukuba, Tsukuba, Ibaraki, Japan.
Hiwasa-Tanase
Kyoko
K
Yoshida
Shigeki
S
Ezura
Hiroshi
H
D002846:Chromatography, Affinity / Q000379:methods*; D006023:Glycoproteins / Q000737:chemistry / Q000302:isolation & purification*; D008670:Metals / Q000737:chemistry; D010940:Plant Proteins / Q000737:chemistry / Q000302:isolation & purification*; D011485:Protein Binding; D013549:Sweetening Agents / Q000737:chemistry / Q000302:isolation & purification*; D032133:Synsepalum / Q000737:chemistry*
D006023:Glycoproteins; D008670:Metals; D010940:Plant Proteins; D013549:Sweetening Agents; C008533:miraculin protein, Synsepalum dulcificum