Lignot B, Froidevaux R, Nedjar-Arroume N, Guillochon D
Laboratoire de Technologie des Substances Naturelles, IUT 'A' Lille 1, BP 179, 59653 Villeneuve d'Ascq Cédex, France.
Biotechnol Appl Biochem. 1999 Feb;29 ( Pt 1):25-30.
The effect of the composition of the solvent on the peptic hydrolysis of bovine haemoglobin was studied to improve the preparation of two opioid peptides. Peptic hydrolysis was performed for 24 h at 23 degrees C in 0.1 M sodium acetate buffer, pH 4.5. A HPLC method was reported for isolating LVV-haemorphin-7 and VV-haemorphin-7 in one step. The kinetics of appearance of haemorphins was investigated in the presence of 20% (v/v) ethanol, a stabilizing solvent of haemoglobin or urea, a denaturant agent. Ethanol improved the yield of VV-haemorphin-7, whereas urea improved the yield of the two haemorphins. Because the amounts of haemorphins observed in urea were greater than in ethanol, the denatured state of haemoglobin is more favourable to obtaining LVV-haemorphin-7 and W-haemorphin-7. The peptide bonds that give rise to haemorphins would be more accessible to the pepsin.
研究了溶剂组成对牛血红蛋白胃蛋白酶水解的影响，以改进两种阿片肽的制备。在0.1M醋酸钠缓冲液（pH 4.5）中于23℃进行胃蛋白酶水解24小时。报道了一种通过高效液相色谱法一步分离LVV-血啡肽-7和VV-血啡肽-7的方法。在存在20%（v/v）乙醇（血红蛋白的一种稳定溶剂）或尿素（一种变性剂）的情况下，研究了血啡肽出现的动力学。乙醇提高了VV-血啡肽-7的产量，而尿素提高了两种血啡肽的产量。由于在尿素中观察到的血啡肽量大于在乙醇中的量，血红蛋白的变性状态更有利于获得LVV-血啡肽-7和W-血啡肽-7。产生血啡肽的肽键对胃蛋白酶来说更容易接近。
