Technische Universität Berlin, Chair of Bioprocess Engineering, Berlin, Germany.
innoFSPEC, University of Potsdam, Potsdam, Germany.
Microb Biotechnol. 2023 Feb;16(2):295-306. doi: 10.1111/1751-7915.14104. Epub 2022 Aug 3.
Bioconversion of waste animal fat (WAF) to polyhydroxyalkanoates (PHAs) is an approach to lower the production costs of these plastic alternatives. However, the solid nature of WAF requires a tailor-made process development. In this study, a double-jacket feeding system was built to thermally liquefy the WAF to employ a continuous feeding strategy. During laboratory-scale cultivations with Ralstonia eutropha Re2058/pCB113, 70% more PHA (45 g L ) and a 75% higher space-time yield (0.63 g L h ) were achieved compared to previously reported fermentations with solid WAF. During the development process, growth and PHA formation were monitored in real-time by in-line photon density wave spectroscopy. The process robustness was further evaluated during scale-down fermentations employing an oscillating aeration, which did not alter the PHA yield although cells encountered periods of oxygen limitation. Flow cytometry with propidium iodide staining showed that more than two-thirds of the cells were viable at the end of the cultivation and viability was even little higher in the scale-down cultivations. Application of this feeding system at 150-L pilot-scale cultivation yielded in 31.5 g L , which is a promising result for the further scale-up to industrial scale.
利用废动物脂肪(WAF)生物转化为聚羟基脂肪酸酯(PHA)是降低这些塑料替代品生产成本的一种方法。然而,WAF 的固体性质需要专门的工艺开发。在这项研究中,构建了一个双层进料系统,将 WAF 热液化,以采用连续进料策略。在实验室规模的 Ralstonia eutropha Re2058/pCB113 培养中,与之前报道的使用固态 WAF 的发酵相比,PHA(45 g/L)产量增加了 70%,时空产率(0.63 g/L·h)提高了 75%。在开发过程中,通过在线光子密度波光谱实时监测了生长和 PHA 形成。通过采用振荡曝气进行放大发酵进一步评估了工艺的稳健性,尽管细胞经历了氧气限制期,但这并没有改变 PHA 的产率。碘化丙啶染色的流式细胞术表明,培养结束时超过三分之二的细胞仍然存活,在缩小规模的培养中,细胞的存活率甚至更高。在 150-L 中试规模培养中应用该进料系统,PHA 的产量为 31.5 g/L,这是进一步放大到工业规模的有希望的结果。