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

虽然加拿大已经种植了这种作物，但如果在出口之前增加更多的附加值，它的经济将会受益。通过生物催化转化，可以为农业产出增加显著价值。然而，到目前为止，由于酶的效率仍然太低，这种工艺不能应用于加拿大大多数价值较低的农产品。克服这一障碍将使我们能够增加这一价值，发展我们的市场，加强我们的经济，创造更多的就业机会，并发展加拿大的企业。在此，我们建议应用最先进的合成生物学技术来开发一种工业生物催化系统，应用于可持续地去除膳食加工过程中不需要的抗营养/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，单宁只会减少摄食和营养吸收。这导致了动物体重的减轻，特别是对单群动物来说。因此，任何含有单宁的生物质或膳食(如豆粕)在饲料中都是有问题的，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)的新兴生物技术将极大地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过当前生物学所能影响的范围。该项目将看到微生物单宁降解途径的生物催化成分将被GCE优化。整合的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。经过GCE优化的衍生生物催化剂将针对不同的生物质进行测试，包括豆粕和油菜籽提取物，以考察它们与其他发酵和酶过程相结合降解单宁的能力。最后，通过体外(细胞和组织)生物测定，将证明衍生产品的营养吸收曲线有所改善。该项目的潜在成果包括关于GCE化学的潜力的知识，从而能够设计出显著更有效的生物催化剂，以及用于从生物质中去除单宁的原型生物催化剂和生物过程。

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.