Coflobiopro: Continuous flow biocatalytic process for production of value-added chemicals using magnetic nanoparticle immobilized enzyme microreactors

Coflobiopro项目将建立一个功能最佳的连续酶级联，以纯甘油为底物，生产乙酰和2,3-丁二醇（2,3bd）作为主要产品。2,3- bd具有极高的工业价值，在航空燃料、橡胶、制药、甲乙基酮（燃料添加剂）、食品（双乙酰）和乙酰生产（挥发性食品添加剂）等领域具有潜在的应用价值。据报道，目前2,3- bd的主要下游产品的全球市场价值为每年430亿美元，其中乙托因的价值约为每吨3万美元。该项目将利用当前流行的技术进步，在单个微反应器平台中包装催化流通过系统。利用生物工程微生物从可再生基质中生成2,3-丁二醇（2,3- bd）已经提出了有效的研究策略，但由于最终产物体积小，成功率有限。为了解决这个问题，我们设计了一种由磁性纳米颗粒（MNP）固定化生物催化剂在填充床酶促微反应器中介导的合成代谢途径。首先，我们将确定研究中每个顺序反应步骤的产品输出的最佳条件。MNP固定化酶醛糖醇氧化酶、二羟基酸脱氢酶、a-乙酰乳酸合酶、a-乙酰乳酸脱羧酶和丁二醇脱氢酶在单个反应器中催化的单个反应将决定产物产量的最佳条件。最后，优化比例的mnp固定化酶将装载在单个反应器中，并在优化条件下连续模式下进行反应。Coflobiopro的新颖之处在于，它结合了功能最佳的MNP固定化酶，在微反应器中以顺序的方式排列，在连续的生物催化级联中产生增值生物燃料。

The Coflobiopro project will establish an optimally functioning continuous enzyme cascade with pure glycerol as substrate to produce acetoin and 2,3-butanediol(2,3BD) as the major products. 2,3-BD has an extremely high industrial value with potential application in the sectors of aviation fuel, rubber, pharmaceuticals, methyl-ethyl ketone (fuel additive), foodstuff(diacetyl) and acetoin production (volatile food additive). The current global market values of the major downstream products from 2,3-BD is reported to be $43 billion per year with the value of acetoin reported to be approximately $30000 per ton. The project will utilize currently prevailing technological advancements in a catalytic flow through system packed in a single microreactor platform. Effective research strategies have been put forth towards generation of 2, 3 butanediol (2,3-BD) by bioengineered microbial species from renewable substrates but with limited success rate since the terminal product is low in volume. To counter this, we have devised a synthetic metabolic pathway mediated by magnetic nanoparticle (MNP) immobilized biocatalysts in a packed bed enzymatic microreactor. Initially, we will ascertain the optimal conditions for product outputs for each of the sequential reaction steps in the study. The individual reactions catalyzed by the MNP immobilized enzymes alditol oxidase, dihydroxy acid dehydratase, a-acetolactate synthase, a-acetolactate decarboxylase and butanediol dehydrogenase in individual reactors will determine the conditions that lends optimal output of the resultant products. Finally, the optimized proportions of the MNP-immobilised enzymes will be loaded in a single reactor and the reactions will be performed under optimized conditions in continuous mode. The novelty of Coflobiopro is that it incorporates optimally functioning MNP immobilized enzymes aligned in a sequential manner in a microreactor for generation ofvalue-added biofuels in a continuous biocatalytic cascade.