正交试验设计优化丝绸水解条件

Optimization of Hydrolysis Conditions of Silk with Orthogonal Experimental Design

为了正确测定丝绸蛋白质的氨基酸含量,采用正交试验设计法对水解条件进行了优化。通过邻苯二甲醛(OPA)与芴甲氧羰碳氯(FMOC-Cl)在线自动柱前衍生的高效液相色谱法,对丝绸蛋白水解液中的氨基酸含量进行检测分析,从而评判各水解条件下丝绸蛋白水解液中氨基酸的含量变化。通过综合优化分析,得到最适合丝绸的水解条件为4 mol/L盐酸水解液,105℃恒温水解15 h。通过对比标准水解条件下与优化后的水解条件下的氨基酸分析结果,除组氨酸、异亮氨酸两种含量较低的氨基酸外,其他氨基酸含量明显高于标准酸条件水解后的氨基酸含量,增幅在5.88%～73.13%,总氨基酸含量增加13.68%,能更准确地反应丝绸的氨基酸含量组成。

In allusion to the protein in silk, this paper optimizes hydrolysis conditions with orthogonal experimental design method ; detects and analyzes the content of amino acid in silk protein hydrolysate with high performance liquid chromatography of OPA and FMOC-Cl online automatic precolumn derivatization and thus evaluates the content change of amino acid in silk protein hydrolysate under various hydrolysis conditions; obtains the hydrolysis condition most suitable tot silk Ihrough comprehensive optimized analysis, i.e. 4 mol/L hydrochloric acid hydrolysate and hydrolysis for 15 h at constant temperature 105 ℃. By eomparing the results of amino acid analysis under the standard hydrolysis condition and optimized hydrolysis condition, we can see that, except histidine and isoleueine with a lower content, other amino acids have a content significantly higher than that after hydrolysis under the standard acid condition; the growing rate is between 5. 88% and 73. 13% and the total content of amino acid increases by 13.68% , which can retlect the amino acid content composition of silk more accurately.