Potential of sugar beet vinasse as a feedstock for biocatalyst production within an integrated biorefinery context
Potential of sugar beet vinasse as a feedstock for biocatalyst production within an integrated biorefinery context
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甜菜酒糟作为综合生物精炼背景下生物催化剂生产原料的潜力
DOI：
10.1002/jctb.5819
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发表时间：
2018
影响因子：
3.4
通讯作者：
Suhaili N
中科院分区：
文献类型：
--
作者：
Suhaili N
背景本工作探讨了酒糟作为综合甜菜生物精炼厂中工业生物催化剂生产的廉价原料的可行性。作为一个例子，研究了大肠杆菌 BL21 中 CV2025 ω-转氨酶 (ω-TAm) 的生产。结果酒糟的表征表明，它主要由甘油以及几种还原糖、糖醇、乙酸酯、多酚和蛋白质组成。初步结果显示E.在使用 17% 至 25% (v/v) 酒糟的培养物中，大肠杆菌BL21 细胞生长和 CV2025 ω-TAm 生产是可行的，且浓度较高，显示出抑制作用。酒糟中存在的 d-半乳糖促进 pQR801 质粒的自诱导，从而无需添加昂贵的异丙基-β-d-硫代半乳糖苷 (IPTG) 即可表达 CV2025 ω-TAm。对不同酒糟预处理方案的评估证实，简单稀释酒糟足以将多酚的浓度降低至抑制水平以下。使用受控 24 孔微生物反应器平台进行的优化实验表明，在稀释酒糟培养基中添加 10 g L−1 酵母提取物可将比生长速率、最大生物量浓度、CV2025 ω-TAm 体积和比活性分别提高 2.8、2.5、5.4 和 3 倍。 E. 代谢偏好的调查当在酒糟中生长时，大肠杆菌BL21 在同时代谢甘油、d-木糖醇、d-卫矛醇和乙酸盐之前表现出优先利用 D-甘露醇。基于匹配的体积传质系数 (kLa)，论证了批量 CV2025 ω-TAm 生产的优化条件在 7.5 L 搅拌釜反应器 (STR) 中的放大。结果显示，细胞生长、底物消耗和 CV2025 ω-TAm 产量具有良好的可比性，代表超过 700 倍的体积尺度平移。当在较高的 kLa 值下运行时，STR 中的 CV2025 ω-TAm 产量可能进一步增强。结论这项工作描述了酒糟在微生物酶生产中的应用前景，以及对复杂原料中碳源利用的见解。利用酒糟作为发酵原料可以进一步扩展到涉及不同微生物和目标酶的其他过程。 © 2018 The Authors.Journal of Chemical Technology & Biotechnology 由 John Wiley & Sons Ltd 代表化学工业协会出版。
BACKGROUNDThis work explores the feasibility of vinasse as an inexpensive feedstock for industrial biocatalyst production within the context of an integrated sugar beet biorefinery. As an exemplar, production of CV2025 ω‐Transaminase (ω‐TAm) inEscherichia coliBL21 was studied.RESULTSCharacterisation of vinasse showed that it comprised mainly of glycerol along with several reducing sugars, sugar alcohols, acetate, polyphenols and protein. Preliminary results showedE. coliBL21 cell growth and CV2025 ω‐TAm production were feasible in cultures using 17% to 25% (v/v) vinasse with higher concentrations demonstrating inhibitory effects. Thed‐galactose present in vinasse facilitated auto‐induction of the pQR801 plasmid enabling CV2025 ω‐TAm expression without addition of expensive Isopropyl‐β‐d‐thiogalactopyranoside (IPTG). Assessment of different vinasse pre‐processing options confirmed simple dilution of the vinasse was sufficient to reduce the concentration of polyphenols to below inhibitory levels. Optimisation experiments, carried out using a controlled, 24‐well microbioreactor platform, showed supplementation of diluted vinasse medium with 10 g L−1yeast extract enabled enhancements of 2.8, 2.5, 5.4 and 3‐fold in specific growth rate, maximum biomass concentration, CV2025 ω‐TAm volumetric and specific activity, respectively. Investigation into the metabolic preferences ofE. coliBL21 when grown in vinasse showed a preference for D‐mannitol utilisation before simultaneous metabolism of glycerol,d‐xylitol,d‐dulcitol and acetate. Scale‐up of optimised conditions for batch CV2025 ω‐TAm production to a 7.5 L stirred tank reactor (STR) was demonstrated based on matched volumetric mass transfer coefficient (kLa). The results showed good comparability with respect to cell growth, substrate consumption and CV2025 ω‐TAm production representing over a 700‐fold volumetric scale translation. Further enhancements in CV2025 ω‐TAm production were possible in the STR when operated at higher kLa values.CONCLUSIONThis work describes the promising application of vinasse for production of microbial enzymes and insights into carbon source utilisation in complex feedstocks. Exploitation of vinasse as a fermentation feedstock could be further extended to other processes involving different microorganisms and target enzymes. © 2018 The Authors.Journal of Chemical Technology & Biotechnologypublished by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.
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