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在一个商业生产项目的实验室实验中,对从嗜盐栖热菌中提取和纯化相容性溶质四氢嘧啶的方法进行优化。

Optimization of the extraction and purification of the compatible solute ectoine from Halomonas elongate in the laboratory experiment of a commercial production project.

作者信息

Chen Ruifeng, Zhu Lijun, Lv Lihuo, Yao Su, Li Bin, Qian Junqing

机构信息

Zhejiang University of Technology, No.18, Chaowang Road, Hangzhou, 310014, Zhejiang, China.

Zhejiang Apeloa Jiayuan Pharmaceutical Co., Ltd, Hengdian Industrial Zone, Dongyang, 322118, Zhejiang, China.

出版信息

World J Microbiol Biotechnol. 2017 Jun;33(6):116. doi: 10.1007/s11274-017-2281-y. Epub 2017 May 9.

Abstract

Optimization of compatible solutes (ectoine) extraction and purification from Halomonas elongata cell fermentation had been investigated in the laboratory tests of a large scale commercial production project. After culturing H. elongata cells in developed medium at 28 °C for 23-30 h, we obtained an average yield and biomass of ectoine for 15.9 g/L and 92.9 (OD), respectively. Cell lysis was performed with acid treatment at moderate high temperature (60-70 °C). The downstream processing operations were designed to be as follows: filtration, desalination, cation exchange, extraction of crude product and three times of refining. Among which the cation exchange and extraction of crude product acquired a high average recovery rate of 95 and 96%; whereas a great loss rate of 19 and 15% was observed during the filtration and desalination, respectively. Combined with the recovering of ectoine from the mother liquor of the three times refining, the average of overall yield (referring to the amount of ectoine synthesized in cells) and purity of final product obtained were 43% and over 98%, respectively. However, key factors that affected the production efficiency were not yields but the time used in the extraction of crude product, involving the crystallization step from water, which spended 24-72 h according to the production scale. Although regarding to the productivity and simplicity on laboratory scale, the method described here can not compete with other investigations, in this study we acquired higher purity of ectoine and provided downstream processes that are capable of operating on industrial scale.

摘要

在一个大规模商业生产项目的实验室测试中,对从长枝盐单胞菌细胞发酵液中提取和纯化相容性溶质(四氢嘧啶)的工艺进行了优化。在28°C下于改良培养基中培养长枝盐单胞菌细胞23 - 30小时后,我们分别获得了平均产量为15.9 g/L的四氢嘧啶和生物量为92.9(OD)。通过在中高温(60 - 70°C)下进行酸处理来进行细胞裂解。下游加工操作设计如下:过滤、脱盐、阳离子交换、粗产品提取和三次精制。其中阳离子交换和粗产品提取的平均回收率分别高达95%和96%;而在过滤和脱盐过程中分别观察到19%和15%的较大损失率。结合从三次精制母液中回收四氢嘧啶,最终获得的平均总产率(指细胞中合成的四氢嘧啶量)和最终产品纯度分别为43%和超过98%。然而,影响生产效率的关键因素不是产率,而是粗产品提取过程中所花费的时间,这涉及从水中结晶的步骤,根据生产规模,该步骤耗时24 - 72小时。尽管就实验室规模的生产率和简便性而言,本文所述方法无法与其他研究竞争,但在本研究中我们获得了更高纯度的四氢嘧啶,并提供了能够在工业规模上运行的下游工艺。

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