为提高5-羟基-L-色氨酸(5-hydroxy-L-tryptophan,5-HTP)提取分离效率,降低能耗和污染排放,选择陶瓷超滤膜应用于从发酵液中提取5-HTP的工艺,对陶瓷超滤膜的过滤条件进行优化.首先优选出适合的膜孔径为10 nm,进一步优化得到陶瓷超滤膜分离提取5-HTP的3个最优工艺参数:发酵液pH值3.5,跨膜压差0.75 MPa,料液过膜温度35 ℃.在此工艺条件下,5-HTP的透过率在92%以上,对游离蛋白质截留率达 90%以上,满足进一步浓缩、结晶制备5-HTP的要求,达到了优化预期效果,为陶瓷超滤膜应用于从发酵液中工业化提取5-HTP提供了可靠依据.
In order to improve the extraction and separation efficiency of 5-hydroxy-L-tryptophan(5-HTP)and reduce energy consumption and pollution emission, ceramic ultrafiltration membrane was selected to extract 5-HTP from fermentation broth. Firstly, the suitable membrane pore size of 10 nm was selected, and then the three optimum process parameters for the separation and extraction of 5-HTP by ceramic ultrafiltration membrane were further optimized: pH value of fermentation solution is 3.5, transmembrane pressure is 0.75 MPa, and transmembrane temperature is 35 ℃. Under this process condition, the penetration ratio of 5-HTP is more than 92%, and the rejection ratio of free protein is more than 90%, which meets the requirements of further concentration and crystallization to prepare 5-HTP, and achieves- DOI:10.3969/j.issn.1000-1565.2024.01.010陶瓷超滤膜在5-羟基-L-色氨酸发酵产物提取中的应用张怡君1,田俊波2,3,杨志彬2,3,郝晋博1,胡江林2,3,吕志堂1(1.河北大学 生命科学学院,河北省微生物多样性研究与应用重点实验室,河北 保定 0710022.河北维达康生物科技有限公司 技术研发中心,河北 保定 072152;3.保定保利瑞合生物科技有限公司 技术研发中心,河北 保定 072152)摘 要:为提高5-羟基-L-色氨酸(5-hydroxy-L-tryptophan,5-HTP)提取分离效率,降低能耗和污染排放,选择陶瓷超滤膜应用于从发酵液中提取5-HTP的工艺,对陶瓷超滤膜的过滤条件进行优化.首先优选出适合的膜孔径为10 nm,进一步优化得到陶瓷超滤膜分离提取5-HTP的3个最优工艺参数:发酵液pH值3.5,跨膜压差0.75 MPa,料液过膜温度35 ℃.在此工艺条件下,5-HTP的透过率在92%以上,对游离蛋白质截留率达 90%以上,满足进一步浓缩、结晶制备5-HTP的要求,达到了优化预期效果,为陶瓷超滤膜应用于从发酵液中工业化提取5-HTP提供了可靠依据.关键词:陶瓷超滤膜;5-羟基-L-色氨酸;过滤条件;透过率;截留率中图分类号:Q81 文献标志码:A 文章编号:1000-1565(2024)01-0077-07Application of ceramic ultrafiltration membrane in the extraction of 5-hydroxy-L-tryptophan from fermentation brothZHANG Yijun1, TIAN Junbo2,3, YANG Zhibin2,3, HAO Jinbo1, HU Jianglin2,3, LYU Zhitang1(1. Key Laboratory of Microbial Diversity Research and Application of Hebei Province, College of Life Sciences, Hebei University,Baoding 071002, China;2. Technology Research and Development Center,Hebei VDK Natura Syn Co. Ltd., Baoding 072152, China;3. Technology Research and Development Center,Baoding BLRH Biotech Co. Ltd., Baoding 072152, China)Abstract: In order to improve the extraction and separation efficiency of 5-hydroxy-L-tryptophan(5-HTP)and reduce energy consumption and pollution emission, ceramic ultrafiltration membrane was selected to extract 5-HTP from fermentation broth. Firstly, the suitable membrane pore size of 10 nm was selected, and then the three optimum process parameters for the separation and extraction of 5-HTP by ceramic ultrafiltration membrane were further optimized: pH value of fermentation solution is 3.5, transmembrane pressure is 0.75 MPa, and transmembrane temperature is 35 ℃. Under this process condition, the penetration ratio of 5-HTP is more than 92%, and the rejection ratio of free protein is more than 90%, which meets the requirements of further concentration and crystallization to prepare 5-HTP, and achieves- 收稿日期:2023-08-28;修回日期:2023-09-06 基金项目:河北省重点研发计划项目(22372403D) 第一作者:张怡君(1996—),女,河北大学在读硕士研究生,主要从事微生物资源与系统学研究.E-mail: zyjdyx898@163.com 通信作者:吕志堂(1973—),男,河北大学教授,博士,主要从事微生物资源与系统学、微生物组学和合成生物学研究.E-mail: lzt325@hbu.edu.cn胡江林(1970—),男,研究员,主要从事制药工程、合成生物学研究.E-mail:hujianglin@vdkbio.com第1期张怡君等:陶瓷超滤膜在5-羟基-L-色氨酸发酵产物提取中的应用the expected effect of the optimization. It provides a reliable basis for the application of ceramic ultrafiltration membrane to industrial extraction of 5-HTP from fermentation broth.
[1] BIRDSALL T C. 5-Hydroxytryptophan: a clinically-effective serotonin precursor[J]. Altern Med Rev, 1998, 3(4): 271-280.
[2] HALFORD J C G, HARROLD J A, BOYLAND E J, et al. Serotonergic drugs: effects on appetite expression and use for the treatment of obesity[J]. Drugs, 2007, 67(1): 27-55. DOI:10.2165/00003495-200767010-00004
[3] HAWKINS M. A 10-year history of using of 5-hydroxytryptophan for severe insomnia in a 15-year-old with autism, seizures, and sleep apnea; cause for concern[J]. Sleep Med, 2019, 64: S148. DOI: 10.1016/j.sleep.2019.11.406
[4] ZHANG H H, ZHAO H T, YANG X X, et al. 5-Hydroxytryptophan, a precursor for serotonin synthesis, reduces seizure-induced respiratory arrest[J]. Epilepsia, 2016, 57(8): 1228-1235. DOI: 10.1111/epi.13430.
[5] WANG H, LIU S Q, LI J, et al. 5-Hydroxytryptophan suppresses the abdominal fat deposit and is beneficial to the intestinal immune function in broilers[J]. Front Physiol, 2020, 11: 655. DOI: 10.3389/fphys.2020.00655.
[6] 刘丽艳,张宏馨,黄珊,等.双波长HPLC-荧光法同时测定褪黑素和维生素B6[J].河北大学学报(自然科学版), 2019, 39(5): 491-497. DOI: 10.3969/j. issn. 1000-1565.2019.05.008.
[7] 张智,谷玮玮,马蕴蕾, 等.褪黑素通过抑制P-选择素表达减轻内毒素致大鼠急性肺损伤[J].河北大学学报(自然科学版), 2016, 36(1): 78-83. DOI: 10. 3969 /j. issn. 1000-1565. 2016. 01. 013.
[8] TURNER E H, LOFTIS J M, BLACKWELL A D. Serotonin a la carte: supplementation with the serotonin precursor 5-hydroxytryptophan[J]. Pharmacol Ther, 2006, 109(3): 325-338. DOI: 10.1016/j.pharmthera.2005.06.004.
[9] LIN Y H, SUN X X, YUAN Q P, et al. Engineering bacterial phenylalanine 4-hydroxylase for microbial synthesis of human neurotransmitter precursor 5-hydroxytryptophan[J]. ACS Synth Biol, 2014, 3(7): 497-505. DOI: 10.1021/sb5002505.
[10] 孙敬方,赵云现.一种生产5-羟色氨酸的方法: CN107267417B[P]. 2018-03-23.
[11] 赵云现,杨志彬,李爱涛,等.一种新的色氨酸羟化酶突变体及其应用: CN112522222B[P]. 2021-12-07.
[12] LI S L, CHOU K S, LIN J Y, et al. Study on the microfiltration of Escherichia coli-containing fermentation broth by a ceramic membrane filter[J]. J Membr Sci, 1996, 110(2): 203-210. DOI: 10.1016/0376-7388(95)00250-2.
[13] TOMCZAK W, GRYTA M. Comparison of polypropylene and ceramic microfiltration membranes applied for separation of 1, 3-PD fermentation broths and Saccharomyces cerevisiae yeast suspensions[J]. Membranes, 2021, 11(1): 44. DOI: 10.3390/membranes11010044.
[14] NIE C Y, LUAN W, CHEN X F, et al. Comparison of ceramic microfiltration and ultrafiltration membranes for the clarification of simulated sebacic acid fermentation broth[J]. J Environ Chem Eng, 2023, 11(3): 109820. DOI: 10.1016/j.jece.2023.109820.
[15] SZCZYGIEŁDA M, PROCHASKA K. Downstream separation and purification of bio-based alpha-ketoglutaric acid from post-fermentation broth using a multi-stage membrane process[J]. Process Biochem, 2020, 96: 38-48. DOI: 10.1016/j.procbio.2020.05.026.
[16] ESCUDERO I, RUIZ M O, BENITO J M, et al. Recovery of α-phenylglycine by micellar extractive ultrafiltration[J]. Chem Eng Res Des, 2006, 84(7): 610-616. DOI: 10.1205/cherd.05217.
[17] WASZAK M, GRYTA M. The ultrafiltration ceramic membrane used for broth separation in membrane bioreactor[J]. Chem Eng J, 2016, 305: 129-135. DOI: 10.1016/j.cej.2015.11.058.
[18] 陈灼湖,李友明,吕阳爱,等.一种采用陶瓷膜微滤分离技术分离螺旋霉素发酵液的方法: CN102351930A[P]. 2012-02-15.
[19] BRADFORD M M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding[J]. Anal Biochem, 1976, 72(1/2): 248-254. DOI: 10.1016/0003-2697(76)90527-3.
[20] JONSSON G. Molecular weight cut-off curves for ultrafiltration membranes of varying pore sizes[J]. Desalination, 1985, 53(1/2/3): 3-10. DOI: 10.1016/0011-9164(85)85048-7.
[21] CALVO J I, PEINADOR R I, PRÁDANOS P, et al. Liquid-liquid displacement porometry to estimate the molecular weight cut-off of ultrafiltration membranes[J]. Desalination, 2011, 268(1/2/3): 174-181. DOI: 10.1016/j.desal.2010.10.016.
[22] HEMM M R, WEAVER J, STORZ G. Escherichia coli small proteome[J]. EcoSal Plus, 2020, 9(1): 10.1128/ecosalplus.ESP-10.1128/ecosalplus0031-2019. DOI: 10.1128/ecosalplus.ESP-0031-2019.
[23] HADGRAFT J W, PRICE S A P, WEST G B. The stability of solutions of 5-hydroxytryptophan[J]. J Pharm Pharmacol, 1958, 10(S1): 87T-91T. DOI: 10.1111/j.2042-7158.1958.tb10386.x.
[24] 张勤,胡雪松,秦岭,等.超滤膜分离纯化加纳籽中5-羟基色氨酸的研究[J].天然产物研究与开发, 2014, 26(12): 2033-2036. DOI: 10.16333/j.1001-6880.2014.12.026.
[25] LI Y B, SHAHBAZI A, KADZERE C T. Separation of cells and proteins from fermentation broth using ultrafiltration[J]. J Food Eng, 2006, 75(4): 574-580. DOI: 10.1016/j.jfoodeng.2005.04.045.
[26] GIFUNI I, LAVENANT L, PRUVOST J, et al. Recovery of microalgal protein by three-steps membrane filtration: Advancements and feasibility[J]. Algal Res, 2020, 51: 102082. DOI: 10.1016/j.algal.2020.102082. (
