Corrigendum to bioprocess intensification: Cases that (don’t) work

Wielen, Luuk A. M. van der; Mussatto, Solange I.; Breugel, J. Van

doi:10.1016/j.nbt.2021.01.002

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“…Technologies of advanced bioprocessing are regarded as key components of the transition to a sustainable bioeconomy (Aguilar et al, 2019 ; Sheldon & Woodley, 2018 ). They involve process intensification as a common target of central importance (Kim et al, 2017 ; Stankiewicz & Moulijn, 2000 ; van der Wielen et al, 2021 ; Woodley, 2017 ). High‐performance bioproduction systems and continuous processing are main pillars of integrated strategies towards bioprocess intensification (Buchholz et al, 2005 ; Woodley, 2020 ; Wu et al, 2021 ).…”

Section: Introductionmentioning

confidence: 99%

Leloir glycosyltransferases enabled to flow synthesis: Continuous production of the natural C‐glycoside nothofagin

Liu

Nidetzky

2021

Biotech & Bioengineering

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C-glycosyltransferase (CGT) and sucrose synthase (SuSy), each fused to the cationic binding module Z basic2 , were co-immobilized on anionic carrier (ReliSorb SP400) and assessed for continuous production of the natural C-glycoside nothofagin. The overall reaction was 3ʹ-C-β-glycosylation of the polyphenol phloretin from uridine 5ʹ-diphosphate (UDP)-glucose that was released in situ from sucrose and UDP.Using solid catalyst optimized for total (∼28 mg/g) as well as relative protein loading (CGT/SuSy = ∼1) and assembled into a packed bed (1 ml), we demonstrate flow synthesis of nothofagin (up to 52 mg/ml; 120 mM) from phloretin (≥95% conversion) solubilized by inclusion complexation in hydroxypropyl β-cyclodextrin. About 1.8 g nothofagin (90 ml; 12-26 mg/ml) were produced continuously over 90 reactor cycles (2.3 h/cycle) with a space-time yield of approximately 11 mg/(ml h) and a total enzyme turnover number of up to 2.9 × 10 3 mg/mg (=3.8 × 10 5 mol/mol). The coimmobilized enzymes exhibited useful effectiveness (∼40% of the enzymes in solution), with limitations on the conversion rate arising partly from external liquid-solid mass transfer of UDP under packed-bed flow conditions. The operational half-life of the catalyst (∼200 h; 30°C) was governed by the binding stability of the glycosyltransferases (≤35% loss of activity) on the solid carrier. Collectively, the current study shows integrated process technology for flow synthesis with coimmobilized sugar nucleotide-dependent glycosyltransferases, using efficient glycosylation from sucrose via the internally recycled UDP-glucose. This provides a basis from engineering science to promote glycosyltransferase applications for natural product glycosides and oligosaccharides.

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