Nature-inspired recycling of a protein mixture into a green fluorescent protein-based hydrogel†

Laura Roset Julià, Sebastian J. Maerkl and Francesco Stellacci
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Abstract

Protein-based materials are biocompatible and have a variety of remarkable properties; consequently, they are finding more and more applications. Nature recycles proteins in multiple ways, ranging from bio-degradation (a slow approach) to fast recycling of protein metabolism. The latter is a wonderful example because a random mixture of proteins gets digested into amino acids (AAs), the fundamental building blocks of proteins. These AAs are then used by cells to produce whichever protein is needed at the time of synthesis. Seen through the lens of recycling, this process transforms a random mixture into something not necessarily present at the start but needed at the moment of recycling. We have recently shown that the process of protein recycling can be performed in vitro and called it NaCRe (Nature Inspired Circular Recycling). In a previous NaCRe proof-of-concept experiment, we started with various protein mixtures but were able to produce only small quantities of recycled protein, in the microgram scale. Here, we show that NaCRe can be used to convert milligrams of a protein mixture containing one of the most common protein materials (silk) into a milligram of an hydrogel made of green fluorescent protein (GFP). We show that in order for NaCRe to be efficient the starting protein mixture must contain a good balance of all AAs and discuss the challenges encountered when scaling up NaCRe.

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在大自然的启发下,将蛋白质混合物回收到基于绿色荧光蛋白的水凝胶中
蛋白质基材料具有生物相容性和各种显著特性,因此其应用越来越广泛。大自然以多种方式回收蛋白质,从生物降解(一种缓慢的方式)到蛋白质新陈代谢的快速回收。后者就是一个很好的例子,因为蛋白质的随机混合物会被消化成氨基酸(AAs),这是蛋白质的基本组成成分。然后,细胞利用这些氨基酸合成所需的蛋白质。从循环的角度来看,这一过程将随机混合物转化为开始时不一定存在,但在循环时需要的东西。我们最近证明,蛋白质循环过程可以在体外进行,并称之为 NaCRe(自然启发的循环再造)。在之前的 NaCRe 概念验证实验中,我们从各种蛋白质混合物入手,但只能产生微克级的少量回收蛋白质。在这里,我们展示了如何利用 NaCRe 将毫克级的蛋白质混合物(其中含有一种最常见的蛋白质材料(蚕丝))转化为毫克级的绿色荧光蛋白(GFP)水凝胶。我们的研究表明,为了使 NaCRe 有效,起始蛋白质混合物必须包含所有 AAs 的良好平衡,并讨论了在扩大 NaCRe 规模时遇到的挑战。
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Correction: Hydrothermal synthesis of ZnZrO x catalysts for CO2 hydrogenation to methanol: the effect of pH on structure and activity. Back cover Inside back cover Direct measurement of PFAS levels in surface water using an engineered biosensor. Statistical optimization of cell-hydrogel interactions for green microbiology - a tutorial review.
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