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Mechanistic studies of enzyme degradation in liquid detergent.

机译：液体洗涤剂中酶降解的机理研究。

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Driven by environmental concerns and a need to differentiate branded products from generic competitors, most laundry detergent products now contain enzymes (protease, amylase, and cellulase). The detergent market is also characterized by an increasing consumer preference for liquid products, which presents a formulation problem: enzymes must be shoehorned into an existing product that contains bleaches, surfactants, and as much as 1% protease. Thus, the goal of this research was to apply modern protein chemistry techniques to build a mechanistic understanding of enzyme activity loss in liquid detergent. Proteolysis, surfactant-induced unfolding, and calcium ion removal are all significant pathways for enzyme degradation.; Proteolysis can be problematic in any detergent formulation containing proteases. Experiments were conducted to measure protease degradation rates and catalytic efficiencies in whole detergent containing various thermodynamic stabilizers and enzyme inhibitors. This work emphasizes the conclusion that proteolysis rates can be slowed by both thermodynamic stabilization and kinetic inhibition.; Our work also examined how surfactants mediate enzyme degradation in detergent formulations. Surfactant monomers bind to proteins in a cooperative manner, causing unfolding and activity loss. Surfactant-induced unfolding is a thermodynamically favorable event, however, the kinetics of the unfolding reaction can be slowed significantly by: (1) modifying conditions to remove protein-surfactant electrostatic attraction, or (2) reducing the amount of surfactant monomer available to interact with the protein (e.g., by using mixed surfactants to lower the critical micelle concentration.); Surfactant aggregates hasten protease and amylase degradation in an indirect manner, by competing with the enzymes for their bound calcium ions. We show that calcium-free enzymes are more susceptible to proteolysis and/or surfactant-induced unfolding. This degradation pathway can be effectively blocked by adding calcium to a detergent formulation or by using surfactants such as ethoxylates that do not interact strongly with Ca2+ ions.
机译：由于对环境的关注以及将品牌产品与一般竞争对手区分开的需求，大多数洗衣粉产品现在都包含酶（蛋白酶，淀粉酶和纤维素酶）。洗涤剂市场的特征还在于，消费者越来越喜欢液态产品，这带来了配方问题：必须将酶塞入含有漂白剂，表面活性剂和多达1％蛋白酶的现有产品中。因此，本研究的目的是应用现代蛋白质化学技术来建立对液体洗涤剂中酶活性损失的机理的理解。蛋白质水解，表面活性剂诱导的解折叠和钙离子的去除都是酶降解的重要途径。在任何含有蛋白酶的洗涤剂配方中，蛋白水解都可能会出现问题。进行实验以测量在包含各种热力学稳定剂和酶抑制剂的整个洗涤剂中蛋白酶的降解速率和催化效率。这项工作强调了一个结论，即热力学稳定和动力学抑制均可降低蛋白水解速率。我们的工作还研究了表面活性剂如何介导洗涤剂配方中的酶降解。表面活性剂单体以协同方式结合蛋白质，导致展开和活性丧失。表面活性剂诱导的展开是热力学上有利的事件，但是，可以通过以下方式显着减慢展开反应的动力学：（1）修改条件以去除蛋白质表面活性剂的静电吸引，或（2）减少可相互作用的表面活性剂单体的量与蛋白质结合（例如，通过使用混合表面活性剂降低临界胶束浓度）；表面活性剂聚集体通过与酶竞争其结合的钙离子，以间接的方式促进蛋白酶和淀粉酶的降解。我们表明，无钙酶对蛋白水解和/或表面活性剂诱导的展开更敏感。通过向洗涤剂配方中添加钙或使用不与Ca2 +离子强烈相互作用的表面活性剂（如乙氧基化物），可以有效地阻止这种降解途径。

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