工程毕赤酵母从 CO2 级联从头生物合成乳蛋白,Green Chemistry

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工程毕赤酵母从 CO2 级联从头生物合成乳蛋白
Green Chemistry ( IF 9.3 ) Pub Date : 2023-07-05 , DOI: 10.1039/d3gc00867c
Xueqin Lv _{1,

2} , Shixiu Cui _{1,

2} , Jie Chen ₃ , Lingrui Wang _{1,

2} , Yanfeng Liu _{1,

2} , Jianghua Li _{1,

2} , Guocheng Du ₂ , Xiaohao Liu ₃ , Jian Chen ₂ , Rodrigo Ledesma-Amaro ₄ , Long Liu _{1,

2,

5}

Affiliation

食物蛋白质是关键营养素，在我们的饮食中发挥着重要作用。然而，食品蛋白的获取主要依靠农产品加工业，造成环境污染、土地占用高、水资源消耗大。在这里，我们描述了一种化学生物级联催化（CBCC）系统，该系统将空间分离的CO ₂热催化与酵母发酵相结合，有效地将CO ₂转化为乳蛋白。首先，通过揭示并打破限速步骤，我们构建了能够高效合成人乳铁蛋白（hLF）的毕赤酵母菌株，产量可达5.4 g L ^-1在生物反应器中，是迄今为止报道的最高滴度。然后，使用CuZnAl催化剂在连续流反应器中由CO _{2稳定合成甲醇。}收集的液相产物由99%的高纯度甲醇水溶液组成，产率为6.7%（mol/mol）。毕赤酵母以热产甲醇为唯一碳源的hLF合成滴度达到56.2 mg L ^-1，甲醇转化率为1.1 mg g ^-1，碳原子转化率为0.17%。此外，开发的 CBCC 还用于生产其他乳蛋白，包括骨桥蛋白和乳白蛋白。这些结果说明了串联化学催化和生物催化将可再生资源转化为食品蛋白质的潜力。这种方法为开发蛋白质制造的可持续替代品铺平了道路，使我们能够满足不断增长的需求。

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Cascaded de novo biosynthesis of lacto-proteins from CO2 by engineered Pichia pastoris

Food proteins are key nutrients, which play an important role in our diet. However, the acquisition of food protein mainly depends on the agroindustry, resulting in environmental pollution, high land usage and the consumption of water resources. Here, we describe a chemo-biocascade catalysis (CBCC) system that combines spatially separated CO₂ thermo-catalysis with yeast fermentation to efficiently convert CO₂ to lacto-proteins. First, by uncovering and breaking the rate limiting step, we constructed a Pichia pastoris strain that efficiently synthesized human lactoferrin (hLF), able to reach a production of 5.4 g L⁻¹ in bioreactor, the highest titer reported so far. Then, CuZnAl catalyst was used to stably synthesize the methanol from CO₂ in a continuous-flow reactor. The collected liquid-phase products consisted of high-purity methanol of 99% in water with a yield of 6.7% (mol/mol). The hLF synthesis titer by P. pastoris using thermally generated methanol as the sole carbon source reached 56.2 mg L⁻¹, with a methanol conversion rate of 1.1 mg g⁻¹, and a carbon atoms conversion rate of 0.17%. Additionally, the developed CBCC was also used to produce other dairy proteins including osteopontin and lactalbumin. These results illustrate the potential of tandem chemocatalysis and biocatalysis for the conversion of renewable resources into food proteins. This methodology paves the way towards the development of sustainable alternatives for protein manufacturing that may allow us to meet the growing demand.

更新日期：2023-07-05

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