TY - JOUR
T1 - Assignment of the disulfide bonds in the sweet protein brazzein
AU - Kohmura, Masanori
AU - Ota, Masafumi
AU - Izawa, Hiroyuki
AU - Ming, Ding
AU - Hellekant, Göran
AU - Ariyoshi, Yasuo
PY - 1996/1/1
Y1 - 1996/1/1
N2 - The thermostable sweet protein brazzein consists of 54 amino acid residues and has four intramolecular disulfide bonds, the location of which is unknown. We found that brazzein resists enzymatic hydrolysis at enzyme/substrate ratios (w/w) of 1:100-1:10 at 35-40°Cfor 24-48 h. Brazzein was hydrolyzed using thermolysin at an enzyme/substrate ratio of 1:1 (w/w) in water, pH 5.5, for 6 h and at 50°C. The disulfide bonds were determined, by a combination of mass spectrometric analysis and amino acid sequencing of cystine-containingpeptides, to be between Cys4-Cys52, Cysl6-Cys37, Cys22-Cys47, andCys26-Cys49. These disulfide bonds contribute to its thermostability.
AB - The thermostable sweet protein brazzein consists of 54 amino acid residues and has four intramolecular disulfide bonds, the location of which is unknown. We found that brazzein resists enzymatic hydrolysis at enzyme/substrate ratios (w/w) of 1:100-1:10 at 35-40°Cfor 24-48 h. Brazzein was hydrolyzed using thermolysin at an enzyme/substrate ratio of 1:1 (w/w) in water, pH 5.5, for 6 h and at 50°C. The disulfide bonds were determined, by a combination of mass spectrometric analysis and amino acid sequencing of cystine-containingpeptides, to be between Cys4-Cys52, Cysl6-Cys37, Cys22-Cys47, andCys26-Cys49. These disulfide bonds contribute to its thermostability.
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U2 - 10.1002/(sici)1097-0282(199604)38:4<553::aid-bip10>3.0.co;2-b
DO - 10.1002/(sici)1097-0282(199604)38:4<553::aid-bip10>3.0.co;2-b
M3 - Article
C2 - 8867215
AN - SCOPUS:0030069420
SN - 0006-3525
VL - 38
SP - 553
EP - 556
JO - Biopolymers
JF - Biopolymers
IS - 4
ER -