草酸水解聚唾液酸分离提纯制备N-乙酰神经氨酸

doi:10.3969/j.issn.0253-2778.2020.05.008

中国科学技术大学学报 ›› 2020, Vol. 50 ›› Issue (5): 605-611.DOI: 10.3969/j.issn.0253-2778.2020.05.008

草酸水解聚唾液酸分离提纯制备N-乙酰神经氨酸

吴志波，陈祥松，吴金勇，孙立洁，袁丽霞，姚建铭

1.中国科学院等离子体物理研究所,安徽合肥 230031;2.淮南新能源研究中心,安徽淮南 232001；3.中国科学技术大学，安徽合肥 230026

收稿日期:2019-04-16 修回日期:2019-11-25 接受日期:2019-11-25 出版日期:2020-05-31 发布日期:2019-11-25
通讯作者: 姚建铭
作者简介:吴志波，男，1992年生，硕士生，研究方向：生物物理学，E-mail：157283617@qq.com
基金资助:
国家自然科学基金（21506235）；安徽省自然科学基金（(1508085MB38）；淮南市重点研究和开发专项（2017A044）；2016年度中科院等离子体物理研究所科学基金（DSJJ-16-YY02）；安徽省面上项目（1808085MB52）资助.

Separation and purification to prepare N-acetylneuraminic acid after oxalic acid hydrolysis of polysialic acid

WU Zhibo, CHEN Xiangsong, WU Jinyong, SUN Lijie, YUAN Lixia, YAO Jianming

1. Institute of plasma physics, Chinese Academy of Sciences(ASIPP), Hefei 230031,China； 2. Huainan new energy research center, Hefei 232001,China； 3. University of Science and Technology of China, Hefei, 230026,China

Received:2019-04-16 Revised:2019-11-25 Accepted:2019-11-25 Online:2020-05-31 Published:2019-11-25

摘要/Abstract

摘要： 为了探究草酸水解聚唾液酸分离提纯制备高纯度N-乙酰神经氨酸的工艺条件，以期简化工艺步骤和降低生产成本；作者重点考察了草酸对聚唾液酸的水解，首先对比了草酸和其他常规水解酸的水解效果，确定了草酸为聚唾液酸的水解酸.然后通过正交实验确定最佳草酸水解操作参数：水解温度为90 ℃、水解时间为3.5 h、水解酸浓度为0.1 mol/L和聚唾液酸稀释倍数为5 倍，在此条件下，聚唾液酸的水解率平均可达94%以上，水解收率可达97%以上.通过实验条件及步骤优化，得到分离提纯制备高纯度N-乙酰神经氨酸的最佳工艺.在该工艺下，红外光谱及HPLC分析证实所得产品中主要成分是N-乙酰神经氨酸，其纯度为98.5%，收率为58.4%.

关键词: 聚唾液酸, N-乙酰神经氨酸, 草酸, 陶瓷膜, 结晶

Abstract: The polysialic acid was obtained from fermentation of escherichia coli, and hydrolyzed to separate, purify and prepare the N-acetylneuraminic acid. In order to study the process conditions of separation and purification to prepare high purity of n-acetylneuraminic acid from the polysialic acid which was hydrolyzed by the oxalic acid, so as to simplify the process and reduce the production cost, the authors investigated emphatically the hydrolysis of the polysialic acid by oxalic acid , contrasted firstly the effects on hydrolysis of polysialic acids between oxalic acids as other conventional hydrolytic acids and determined oxalic acids as hydrolytic acids. The best operating parameters of the oxalic acid hydrolysis of the polysialic acid was determined by the orthogonal experiment: hydrolysis temperature 90 ℃, hydrolysis time 3.5 h, hydrolysis acid concentration 0.1 mol/L and diluted multiples of polysialic acid 5 times. In these conditions, the hydrolyze rate of the polysialic acid was more than 94 %, and the hydrolyze yield was more than 97 %. The separation and purification conditions were optimized through experiments, and the separation and purification process for the preparation of high purity N-acetylneuraminic acid was obtained. In these process conditions, the analytic results of the infrared spectrum and HPLC showed that the purity of the N-acetylneuraminic acid product was 98.5 %, and the yield was 58.4 %.

Key words: polysialic acid, N-acetylneuraminic acid, oxalic acid, hydrolysis, separation and purification

中图分类号:

TQ46

吴志波，陈祥松，吴金勇，孙立洁，袁丽霞，姚建铭. 草酸水解聚唾液酸分离提纯制备N-乙酰神经氨酸[J]. 中国科学技术大学学报, 2020, 50(5): 605-611.

WU Zhibo, CHEN Xiangsong, WU Jinyong, SUN Lijie, YUAN Lixia, YAO Jianming. Separation and purification to prepare N-acetylneuraminic acid after oxalic acid hydrolysis of polysialic acid[J]. Journal of University of Science and Technology of China, 2020, 50(5): 605-611.

参考文献

［1］
REVILLA-NUIN B, REGLERO A, FERRERO M A, et al. Uptake of N-acetyl-D-mannosamine: an essential intermediate in polysialic acid biosynthesis by Escherichia coli K92[J]. FEBS Letters, 1999, 449: 183-186.
[2] BRUSS J L, RUTISHAUSER U. Roles, regulation, and mechanism of acid function during neural development[J]. Biochimie, 2001, 83: 635-643.
[3] BLIX F G, GOTTSCHALK A, KLENK E. Proposed nomenclature in the field of neuraminic and sialic acid[J]. Nature, 1957, 179:1088.
[4] SHEN G J, DATTA A K, IZUMI M, et al. Expression of α2, 8/2, 9-Polysialyltransferase from Escherichia coli K92: Characterization of the enzyme and its reaction products[J]. Journal of Biological Chemistry, 1999, 274(49):35139-35146.
[5] 刘景, 任婧, 刘翠平, 等. 动物乳及婴幼儿配方奶中唾液酸的研究及应用进展[J]. 食品研究与开发, 2013, 34(11): 105-108.
[6] WITCZAK Z J, KARL A N. Carbohydrate in Drugs Design[M]. New York: Marcel Dekker, 1997.
[7] 高剑峰, 冯万祥. 血液中唾液酸的提取和结晶纯化[J]. 中国医药工业杂志, 1996, 27(6): 246-247.
[8] JUNEJA L R, KOKETSU M, NISHIMOTO K, et al. Large-scale preparation of sialic acid from chalaza and egg-yolk membrane[ J]. Carbohydrate Research, 1991, 214:179-186.
[9] WHITEHOUSE M W, ZILLIKEN F. Isolation and determination of neuraminic(sialic) acids[J]. Methods of Biochemical Analysis, 1960, 8: 199-218.
[10] MASAHARU S, YUKIO U, ICHIROU M, et al. Process for manufacturing sialic acids-containing composition: US5270462[P/OL]. 1993-12-14[2019-03-15]. https://europepmc.org/article/PAT/US5270462.
[11] 赵慧, 詹晓北, 朱一晖，等. 聚唾液酸的水解与唾液酸的纯化[J]. 无锡轻工业大学学报, 2004, 23(4): 16-19.
[12] 李文强. 唾液酸发酵控制及其分离纯化的研究[D]. 无锡：江南大学，2005.
[13] 郁丹凤. 聚唾液酸和唾液酸提取工艺的研究[D]. 无锡：江南大学，2008.
[14] 吴金勇, 陈祥松，袁丽霞，等. 磁场条件下Fe2+对产聚唾液酸大肠杆菌的诱变选育[J]. 河南师范大学学报, 2018, 46(6): 80-85.
[15] SVENNERHOLM L. Quantitative estimation of sialic acids：II. A colorimetric resorcinol-hydrochloric acid method[J]. Biochimica et Biophysica Acta, 1957, 24(4): 604-611.

（上接第580页）

[2] PAZY A. Semigroups of Linear Operators and Applications to Partial Differential Equations[M]. New York: Springer, 1983.
[3] ROLEWICZ S. On uniform N-equistability[J]. J Math Anal Appl, 1986, 115(2): 434-441.
[4] PREDA C. On the uniform exponential stability of linear skew-product semiflows[J]. J Funct Spaces Appl, 2006, 4(2): 145-161.
[5] HAI P V. Continuous and discrete characterizations for the uniform exponential stability of linear skew-evolution semiflows[J]. Nolinear Anal, 2010, 72(12): 4390-4396.
[6] PREDA C, PREDA P, BTRAN F. An extension of a theorem of R. Datko to the case of (non)uniform exponential stability of linear skew-product semiflows[J]. J Math Anal Appl, 2015, 425(2): 1148-1154.
[7] PREDA C, ONOFREI O R.Nonuniform exponential dichotomy for linear skew-product semiflows over semiflows[J]. Semigroup Forum, 2018, 96(2): 241-252.
[8] MEGAN M, SASU A L, SASU B. Perron conditions for uniform exponential expansiveness of linear skew-product flows[J].Monatsh Math, 2003, 138(2): 145-157.
[9] MEGAN M, SASU A L, SASU B. Exponential instability of linear skew-productsemiflows in terms of Banach function spaces[J]. Results Math, 2004, 45(3): 309-318.
[10] MEGAN M, SASU A L, SASU B. Exponential stability and exponential instability for linear skew-product flows[J]. Math Bohem, 2004, 129(3): 225-243.
[11] 岳田，雷国梁，宋晓秋.线性斜演化半流一致指数膨胀性的若干刻画[J].数学进展, 2016, 45(3): 433-442.
[12] PREDA P, POGAN A, PREDA C.Functionals on function and sequence spaces connected with the exponential stability of evolutionary processes[J]. Czechoslovak Math, 2006, 131(56): 425-435.

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草酸水解聚唾液酸分离提纯制备N-乙酰神经氨酸

Separation and purification to prepare N-acetylneuraminic acid after oxalic acid hydrolysis of polysialic acid

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