Effect of flux and shear rate on E. coli recovery in tangential flow filtration through a single hollow fiber

IF 2.5 3区 生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Biotechnology Progress Pub Date : 2024-02-08 DOI:10.1002/btpr.3432
Jessica Zuponcic, Fernanda Cunha, Grant Springer, Eduardo Ximenes, Michael R. Ladisch
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Abstract

Pathogenic bacteria which enter a viable but non-culturable (VBNC) state impede efforts to reach detectable concentrations required for PCR methods. This motivated a strategy for tangential flow filtration to concentrate bacteria in aqueous samples while maintaining the bacteria in a viable state, maximizing their recovery and achieving high fluxes through a single hollow fiber membrane. Filtrations were carried out for green fluorescent protein (GFP) E. coli at high shear rates (up to 27,000 sec−1) through 0.2 μm cut-off polyethersulfone (PES) microfilter membranes or 50 kDa polysulfone (PS) ultrafilter membranes. High shear minimized bacterial attachment on membrane surfaces, which would otherwise occur due to forced convection of the particles to the membrane surface at high flux conditions. Single fiber filter modules were constructed to facilitate concentration of Escherichia coli at fluxes ranging from 55 to 4500 L m−2 h−1. The effect of high shear rates on bacterial viability was found to be minimal with bacterial losses during filtration caused principally by their accumulation on the membrane surface. Recoveries of 90% were achievable at high shear rates when the average flux was ≤300 L m−2 h−1. This corresponded to a 3-h filtration time for a 225 mL sample through a single hollow fiber. Detectable bacteria concentrations of 1800 colony-forming unit (CFU)/mL were achieved for starting concentrations of 140 CFU/mL.

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流量和剪切率对单根中空纤维切向流过滤中大肠杆菌回收率的影响。
病原菌进入可存活但不可培养(VBNC)的状态,阻碍了达到 PCR 方法所需的可检测浓度的努力。这就促使我们采用切向流过滤策略来浓缩水样中的细菌,同时保持细菌的存活状态,最大限度地提高细菌回收率,并通过单个中空纤维膜实现高通量。绿色荧光蛋白(GFP)大肠杆菌在高剪切速率(高达 27,000 秒-1)下通过 0.2 μm 截流聚醚砜(PES)微滤膜或 50 kDa 聚砜(PS)超滤膜进行过滤。高剪切力最大程度地减少了细菌在膜表面的附着,否则在高通量条件下,颗粒会被迫对流到膜表面。构建了单纤维过滤模块,以便在 55 至 4500 L m-2 h-1 的通量条件下浓缩大肠杆菌。研究发现,高剪切率对细菌存活率的影响很小,过滤过程中细菌的损失主要是由于它们在膜表面的积聚。当平均通量≤300 L m-2 h-1时,高剪切率下的回收率可达90%。这相当于 225 mL 样品通过一根中空纤维的过滤时间为 3 小时。起始浓度为 140 CFU/mL 时,可检测到的细菌浓度为 1800 菌落总数 (CFU)/mL。
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来源期刊
Biotechnology Progress
Biotechnology Progress 工程技术-生物工程与应用微生物
CiteScore
6.50
自引率
3.40%
发文量
83
审稿时长
4 months
期刊介绍: Biotechnology Progress , an official, bimonthly publication of the American Institute of Chemical Engineers and its technological community, the Society for Biological Engineering, features peer-reviewed research articles, reviews, and descriptions of emerging techniques for the development and design of new processes, products, and devices for the biotechnology, biopharmaceutical and bioprocess industries. Widespread interest includes application of biological and engineering principles in fields such as applied cellular physiology and metabolic engineering, biocatalysis and bioreactor design, bioseparations and downstream processing, cell culture and tissue engineering, biosensors and process control, bioinformatics and systems biology, biomaterials and artificial organs, stem cell biology and genetics, and plant biology and food science. Manuscripts concerning the design of related processes, products, or devices are also encouraged. Four types of manuscripts are printed in the Journal: Research Papers, Topical or Review Papers, Letters to the Editor, and R & D Notes.
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