Effect of molecular patch modification on the stability of dynamic high-pressure microfluidization treated trypsin
Effect of molecular patch modification on the stability of dynamic high-pressure microfluidization treated trypsin
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分子贴片修饰对动态高压微流化处理胰蛋白酶稳定性的影响
DOI：
10.1016/j.ifset.2012.08.001
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发表时间：
2012-10-01
影响因子：
6.6
通讯作者：
Wan, Jie
中科院分区：
文献类型：
--
作者：
Liu, Wei;Zhang, Zhao-Qin;Wan, Jie
以往的研究表明，动态高压微射流（DHPM）处理引起的胰蛋白酶去折叠增强了其水解稳定性。然而，解折叠的胰蛋白酶处于亚稳态，随后将聚集到最低能量状态。本研究选用单甲氧基聚乙二醇-琥珀酰亚胺碳酸酯（mPEG-SC）作为修饰剂，对胰蛋白酶进行修饰，使其作为分子补片。结果表明，mPEG-SC对未折叠胰蛋白酶的贮存稳定性和热稳定性有显著影响。与在100 MPa压力下处理的胰蛋白酶DHPM相比，进一步的mPEG-SC修饰分别在4 ℃ 8天和55 ℃ 10 min后将活性从50%提高到74%和从70%提高到87%。荧光猝灭证明了mPEG-SC与胰蛋白酶的结合，表明分子补丁修饰可以抑制未折叠的胰蛋白酶的复性。产业相关性：动态高压微射流技术（DHPM）是一种很有前途的技术，已被广泛应用于功能特性或结构修饰的创新食品。胰蛋白酶已广泛应用于现代食品工业和生物领域。无论是DHPM处理还是mPEG-SC修饰都能提高胰蛋白酶的热稳定性，而两者联合处理则能更显著地提高胰蛋白酶的热稳定性和储存稳定性。希望本研究能为酶改性的理论和应用基础研究提供新的观点和技术方案。(C)2012爱思唯尔有限公司保留所有权利。
Previous research indicated that the unfolding of trypsin induced by dynamic high-pressure microfluidization (DHPM) treatment had enhanced its hydrolytic stability. However, the unfolding trypsin is in a metastable state and will subsequently aggregate to be in the lowest energy state. In this study, monomethoxy polyethylene glycol-succinimidyl carbonates (mPEG-SC) was chosen to modify trypsin acted as a molecular patch. The results indicated mPEG-SC showed a significant effect on storage stability and thermal stability of unfolded trypsin. Compared to trypsin DHPM-treated under pressure of 100 MPa, further mPEG-SC modification increased the activity from 50% to 74% and from 70% to 87% after 4 degrees C for 8 days and 55 degrees C for 10 min, respectively. Fluorescence quenching demonstrated the conjugation of mPEG-SC and trypsin, and indicated molecular patch modification could inhibit the refolding of unfolded trypsin.Industrial relevance: Dynamic high-pressure microfluidization (DHPM) is a promising technology that has been widely applied for innovative foods with modified functional properties or structure. Trypsin has been widely used in modern food industry and biological field. Either DHPM treatment or mPEG-SC modification enhanced the thermal stability of trypsin, while, the combination of the two enhanced the thermal and storage stability of trypsin more significantly. It is expected that this study can provide a new viewpoint and technical scheme for basic research on theory and application of enzyme modification. (C) 2012 Elsevier Ltd. All rights reserved.