Application of genetic code expansion for development of biocatalysts to enable value-added bioprocessing

虽然加拿大已经种植这种作物，但如果在出口前增加更多的价值，其经济将受益。 生物催化转化可为农业产出增加大量价值。然而，到目前为止，由于酶的效率仍然太低，成本效益比阻碍了这种方法在加拿大大多数低价值农产品上的应用。克服这一障碍将使我们能够增加这一价值，扩大我们的市场，加强我们的经济，创造更多的就业机会和发展加拿大的企业。在此，我们建议应用最先进的合成生物学技术来开发工业生物催化系统，用于在膳食加工过程中可持续地去除不期望的抗营养/适口性因素。事实上，在生命短暂、体重增加为王的畜牧业方面，单宁只会减少喂养和营养吸收。这导致动物体重减轻，特别是对于单胃动物。 因此，含有单宁的任何生物质或粗粉（例如豆类粗粉）在饲料中是有问题的，限制了它们的适用性和价值。 然而，一种名为“遗传密码扩展”（GCE）的新兴生物基技术，正准备大大扩展生物催化剂精密工程的潜力，其特异性，稳定性和效率远远超出当前生物学的影响。该项目将看到微生物单宁降解途径的生物催化成分，以GCE为优化目标。结合合成生物化学将测试使用GCE来增强酶的催化活性和稳定性的潜力。衍生的GCE优化的生物催化剂将针对各种生物质（包括豆类粉和油菜籽提取物）进行测试，以确定其与其他发酵和酶促过程相结合降解单宁的能力。最后，将通过体外（细胞和组织）生物测定证明衍生产品的营养吸收特性得到改善。 该项目的潜在成果包括了解GCE化学的潜力，以使工程的更有效的生物催化剂，以及原型生物催化剂和生物工艺，从生物质中去除单宁。

While Canada already grows the crops, its economy would benefit if more value is added before export. Significant value can be added to agricultural outputs by biocatalytic conversion. However, the cost-to-benefit ratio has up until now, precluded application of such processes to most of Canada's lower value agricultural commodities because enzymes remain too inefficient. Overcoming this hurdle will allow us to add that value, growing our markets, strengthening our economy, creating more jobs and growing Canadian businesses. Herein, we propose to apply state-of-the-art synthetic biology technology to develop an industrial biocatalytic system for application to the sustainable removal of undesirable anti-nutritional/palatability factors during the processing of meals. Indeed, on the side of animal husbandry, where life is short and weight gain is king, tannins serve only to reduce feeding and nutrient absorption. This leads to reduced animal weights, particularly for monogastrics. As such any biomass or meals which contain tannins (such as pulse meals) are problematic in feed, limiting their applicability and value. However, an emerging bio-based technology termed 'genetic code expansion' (GCE), is poised to dramatically expand the potential of precision engineering of biocatalysts with a breadth of specificities, stabilities and efficiencies far beyond what current biology can affect. This project will see biocatalytic components of microbial tannin degradation pathways targeted for optimization by GCE. Incorporated synthetic biochemistries will test the potential to use GCE to enhance catalytic activities and stability of the enzymes. Derived GCE-optimized biocatalysts will be tested against various biomass, including pulse meal and canola extracts, for their ability to degrade tannins in combination with other fermentative and enzymatic processes. Finally, improved nutrient absorption profiles of the derived products will be demonstrated by in vitro (cell and tissue) bioassays. Potential outcomes of this project include knowledge about the potential of GCE chemistries to enable engineering of significantly more efficient biocatalysts as well as prototype biocatalysts and bioprocesses for the removal of tannins from biomass.