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

这项小型企业创新研究(SBIR)第一阶段提案的更广泛影响是开发了一种新的碳利用技术，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。酶法碳利用因其能够从二氧化碳中提取高价值的天然产物而备受关注。酶法碳利用技术的障碍是在无细胞的工业条件下酶和输入分子的寿命都很短。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为基本产品供应链释放二氧化碳作为一种新的自然资源，这些供应链是碳负和水和土地中性的，从而打开一个价值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产业，并促进美国的制造业就业。这项技术能够以比现在的生产成本更低的成本，可持续地、国内地从废旧二氧化碳排放中生产美国最基本的材料。该项目涉及一种生产方法，该方法应用了一种生物启发的非共价酶稳定策略，在工业相关的时间段内，在二氧化碳转化为纺织品的过程中，将碳排放无细胞地转化为用于纺织品的纤维纤维。所提出的稳定方法在不损害基线活性的情况下保留了酶的构象，以允许在非自然环境中的酶活性。利用标准的可逆加成-断裂链转移(RAFT)聚合方法合成杂化聚合物，生成具有不同电荷和疏水/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局部表面结合能够稳定酶，保持其构象，并防止变性或降解。项目范围涵盖关键技术障碍：(1)开发用于碳固定和合成酶的酶稳定技术，(2)开发非自然工业环境中的碳转化系统和工艺，以及(3)确认可从毫升到50L规模的完整概念验证解决方案。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NSF的法定使命，并通过使用基金会的智力优势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.