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，但如果在出口前增加更多的价值，加拿大的经济将受益。通过生物催化转化可以显著增加农业产出的价值。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，促进加拿大企业的发展。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，用于可持续地去除食品加工过程中不良的抗营养/适口因素。事实上，在畜牧业方面，生命短暂，体重增加为王，单宁只会减少饲养和营养吸收。这导致动物体重减少，特别是对于单配偶制。因此，任何含有单宁的生物质或饲料（如脉冲饲料）在饲料中都是有问题的，限制了它们的适用性和价值。然而，一种新兴的基于生物的技术被称为“遗传密码扩展”（GCE），它将极大地扩大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所能影响的范围。本项目将看到微生物单宁降解途径的生物催化成分，以GCE为目标进行优化。结合合成生物化学将测试使用GCE提高酶的催化活性和稳定性的潜力。衍生的gce优化生物催化剂将针对各种生物质进行测试，包括脉冲粉和菜籽油提取物，以测试它们与其他发酵和酶促过程结合降解单宁的能力。最后，将通过体外（细胞和组织）生物测定来证明衍生产品的营养吸收谱的改善。该项目的潜在成果包括GCE化学的潜力，使工程更有效的生物催化剂以及原型生物催化剂和生物过程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.