I-Corps: Enzymes for on-site generation of dilute peracetic acid

稀过氧乙酸(PAA)是食品和饮料行业以及医疗设施和设备的有效消毒剂。预计2018年全球过氧乙酸市场规模为6亿美元。使用PAA的挑战在于，制造商需要创建集中的PAA解决方案。这些对运输、储存和稀释使用都是危险的。建议的解决方案是现场制作PAA，这消除了运输，只有在需要时才制作PAA，这消除了存储，并且只制作稀释的PAA，这就消除了工人的处理步骤。这一解决方案的关键是从更安全的成分--醋酸和过氧化氢--中进行的现场酶催化反应。为了达到这一应用目标，酶需要比现在更稳定，这样它们才能重复使用。过氧乙酸(PAA)是一种强烈的活性氧化剂，使PAA能够杀死广泛的微生物。这个i-Corps团队的目标是在终端用户的现场设施中，通过醋酸和过氧化氢的过水解反应，使用酶来生产PAA。先前的研究发现了高效的酶，每一种酶分子每秒产生10个PAA分子。快速酶催化的PAA的形成创造了机会，使其在应用位置仅根据需求和所需的量进行生产。为了使其在商业上可行，建议的方法是在填充床反应器中使用酶，加入醋酸和过氧化氢，并生产所需数量的PAA。为了实现这一点，需要设计更耐PAA氧化的过水解酶，固定化稳定的酶以实现可重复使用，并优化连续流动反应器的反应条件。目前的提案寻求将酶法生产的PAA商业化，以便更安全地使用，从而检验更广泛的假设，即生物技术微型工厂可以取代传统化工厂。

Dilute peracetic acid (PAA) is a potent disinfectant for the food and beverage industry and for medical facilities and equipment. The estimated global market of peracetic acid in 2018 is US$600 million. The challenge with using PAA is that manufacture creates concentrated PAA solutions. These are hazardous to transport, to store and to dilute for use. The proposed solution is to make PAA on-site, which eliminates transport, make PAA only when needed, which eliminates storage, and make PAA only in the dilute less-hazardous form, which eliminates handling steps by workers. The key to this solution is an enzyme catalyzed reaction on-site from safer ingredients - acetic acid and hydrogen peroxide. To reach this application goal, the enzymes need to be more stable than they are now so they can be reused.Peracetic acid (PAA) is a strong, reactive oxidant which allows PAA to kill a broad range of microbes. This I-Corps team's goal is to produce PAA at end-user's on-site facility using enzymes by the perhydrolysis reaction of acetic acid and hydrogen peroxide. Prior research identified efficient enzymes which creates ten molecule of PAA per second for each molecule of enzyme. The fast enzyme catalyzed formation of PAA created the opportunity to make it at the application location only on demand and also in amounts needed. To make it commercially viable, the proposed approach is to use enzyme in a packed-bed reactor, with acetic acid and hydrogen peroxide fed in and produce PAA with the amounts needed. To achieve this demonstration, it requires engineering of perhydrolase enzymes to be more resistant for oxidation by PAA, immobilization of stabilized enzymes for the reusability and optimize reaction conditions for a continuous flow reactor. The current proposal seeks to commercialize the enzymatic production of PAA for safer use and thereby tests the broader hypothesis that the biotechnology micro factories can replace traditional chemical factories.