SBIR Phase I: Carbon dioxide (CO2) fixation into cellulosic textile products through heteropolymer stabilization of enzymes in cell-free synthesis

这项小企业创新研究（SBIR）第一阶段提案的广泛影响在于开发一种新的碳利用技术，该技术能够从二氧化碳（CO2）中生产高价值的基本产品，如纤维素纺织品、食品和建筑材料。酶促碳利用是高度感兴趣的，因为它能够从二氧化碳中制造高价值的天然产物。在无细胞的工业条件下，酶和输入分子的寿命都很短，这是酶碳利用技术的障碍。该项目将为碳固定和合成酶提供一种新的酶稳定方法。由此产生的系统可能会释放二氧化碳，使其成为碳负、水和土地中性的基本产品供应链的一种新的自然资源，从而释放10万亿美元的全球市场。这种可扩展的技术可以创造一个新的碳工业，并在美国增加制造业就业机会。这项技术可以使美国最重要的材料以比现在更低的成本在国内可持续地从二氧化碳排放中生产出来。该项目涉及一种生产方法，该方法采用生物启发的非共价酶稳定策略，在工业相关时期内，在二氧化碳到纺织品的过程中，将无细胞的碳排放转化为纺织品的纤维素纤维。所提出的稳定方法保留了酶的构象，而不损害基线活性，以允许非自然环境中的酶活性。采用标准的可逆加成-断裂链转移（RAFT）聚合方法合成杂多聚物，产生具有不同电荷和亲疏水特性的不同单体的无序聚合链，以补充和稳定酶表面。聚合物的无序性质使得局部表面结合能够稳定酶，保持其构象并防止其变性或降解。项目范围涵盖了关键的技术障碍：(1)开发用于碳固定和合成酶的酶稳定技术，(2)在非自然工业环境中开发碳转化系统和过程，以及(3)确认一个完整的概念验证解决方案，可以从mL扩展到50L规模。如果成功，结果是一种有效的酶稳定方法，可以在无细胞条件下固定碳排放，用于工业供应链。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

The broader impact of this Small Business Innovation Research (SBIR) Phase I proposal is in the development of a new carbon utilization technology with the ability to make high-value, essential products from carbon dioxide (CO2) like cellulosic textiles, food, and building materials. Enzymatic carbon utilization is of high interest due to its ability to make high-value natural products from carbon dioxide. The impediment to enzymatic carbon utilization technology has been the short longevity of both enzymes and input molecules under cell-free industrial conditions. This project will yield a novel enzyme stabilization approach for carbon fixation and synthesis enzymes. The resulting system may unlock CO2 as a new, natural resource for essential product supply chains that are carbon-negative and water- and land-neutral, unlocking a $10 trillion global market. This scalable technology could create a new carbon industry and advance manufacturing jobs in the United States. This technology could enable sustainable and domestic production of America’s most essential materials from waste carbon dioxide emissions at a cheaper cost than today’s production.This project involves a production method that applies a bio-inspired, non-covalent enzyme stabilization strategy that employs cell-free conversion of carbon emissions into cellulosic fibers for textiles in a CO2-to-textile process over industrially relevant time periods. The proposed stabilization method preserves enzyme conformation without harming baseline activity to allow for enzymatic activity in non-natural environments. The heteropolymers are synthesized using a standard reversible addition-fragmentation chain transfer (RAFT) polymerization approach to produce disordered polymeric chains with diverse monomers of varying charges and hydrophilicity/hydrophobicity characteristics to complement and stabilize enzymes surfaces. The disordered nature of the polymers enables local surface binding to stabilize the enzyme, maintaining its conformation and protecting against denaturing or degradation. The project scope covers key technical hurdles: (1) the development of a enzyme stabilization technique for carbon fixation and synthesis enzymes, (2) the development of a carbon conversion system and process in a non-natural, industrial environment, and (3) confirmation of a full proof-of-concept solution that can be scalable from mL to 50L scale. If successful, the result is an effective enzyme stabilization method that can fix carbon emissions under cell-free conditions for use in industrial supply chains.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.