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超声辅助醇/盐双水相体系从重组马克斯克鲁维酵母中提取虾青素的强化回收。

Enhanced recovery of astaxanthin from recombinant Kluyveromyces marxianus with ultrasonication-assisted alcohol/salt aqueous biphasic system.

机构信息

Faculty of Applied Sciences, UCSI University, UCSI Heights, 56000 Cheras, Kuala Lumpur, Malaysia.

Graduate Institute of Integrated Medicine, College of Chinese Medicine, China Medical University, Taichung 40402, Taiwan; Chinese Medicine Research and Development Center, China Medical University Hospital, Taichung 40447, Taiwan; Graduate Institute of Natural Products, College of Pharmacy, Kaohsiung Medical University, Kaohsiung 80708, Taiwan.

出版信息

J Biosci Bioeng. 2021 Nov;132(5):513-518. doi: 10.1016/j.jbiosc.2021.07.004. Epub 2021 Sep 1.

Abstract

Microbial astaxanthin with strong antioxidant activity is greatly demanded for diverse applications. Extractive disruption in aqueous biphasic system (ABS) integrates the cells disruption and biomolecules recovery processes in one-step operation, allowing the direct recovery of intracellular biomolecules with biphasic system upon released from cells. In this study, astaxanthin was recovered from recombinant Kluyveromyces marxianus yeast cells via extractive disruption using alcohol/salt ABS. Recombinant K. marxianus yeast is engineered to produce high concentration of free form astaxanthin. Highest partition coefficient (K = 90.02 ± 2.25) and yield (Y = 96.80% ± 0.05) of astaxanthin were obtained with ABS composed of 20% (w/w) 1-propanol and 20% (w/w) sodium citrate of pH 5, 0.5% (w/w) yeast cells loading and additional of 1% (w/w) 1-butyl-3-methylimidazolium tetrafluoroborate (Bmim)BF to improve the migration of astaxanthin to alcohol-rich top phase. The incorporation of 2.5 h of ultrasonication to the biphasic system further enhanced the astaxanthin recovery in ABS. The direct recovery of astaxanthin from recombinant K. marxianus cells was demonstrated with the ultrasonication-assisted alcohol/salt ABS which integrates the extraction and concentration of astaxanthin in a single-step operation.

摘要

具有强抗氧化活性的微生物虾青素在多种应用中需求量很大。双水相体系(ABS)中的萃取破坏将细胞破坏和生物分子回收过程集成在一步操作中,允许在从细胞释放后直接从双水相体系中回收细胞内生物分子。在这项研究中,通过使用醇/盐 ABS 从重组马克斯克鲁维酵母细胞中萃取破坏来回收虾青素。重组 K.marxianus 酵母被设计用于生产高浓度的游离虾青素。在 pH 5、0.5%(w/w)酵母细胞负载和额外添加 1%(w/w)1-丁基-3-甲基咪唑四氟硼酸盐(Bmim)BF 以改善虾青素向富含醇的上相迁移的条件下,用由 20%(w/w)1-丙醇和 20%(w/w)柠檬酸钠组成的 ABS 中,虾青素获得了最高的分配系数(K=90.02±2.25)和产率(Y=96.80%±0.05)。将超声处理 2.5 小时加入到双水相体系中进一步提高了 ABS 中虾青素的回收率。超声辅助醇/盐 ABS 直接从重组 K.marxianus 细胞中回收虾青素,该 ABS 将虾青素的提取和浓缩集成在一步操作中。

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