Separation of phenyllactic acid from transformation broth by anion exchange poly(2-hydroxyethyl methacrylate) composite cryogel embedded with nanogels

doi:10.11949/0438-1157.20200623

Abstract

Abstract:

Hydrophobic nanogels were synthesized by emulsion polymerization method under ultrasound using butyl methacrylate as the monomer. Then, anion exchange nano-cryogels embedded with the obtained nanogels were prepared by cryo-polymerization using poly(2-hydroxyethyl methacrylate) as the monomer, followed by a graft polymerization with the monomer (vinylbenzyl) trimethyl ammonium chloride. The anion exchange nano-cryogels were used to isolate phenyllactic acid from biotransformation broth. The results show that the obtained nanocrystalline gel has good permeability and axial dispersion performance. When the flow rate is 1 cm·min^-1, the adsorption capacity of different ratios of nanocrystalline gel for bovine serum protein is 3.9—5.4 mg·ml^-1. When the nano-cryogel with mass ratio of 8∶2(HEMA∶pBMA) was used to isolate phenyllactic acid from transformation broth, phenyllactic acid with high purity of 89.24% and recovery of 93.74% was obtained, indicating that the nano-cryogels could have potential applications in bioseparation.

Key words: nanogels, nano-cryogels, adsorption, phenyllactic acid

摘要：

采用超声乳化聚合法制备了聚甲基丙烯酸丁酯纳凝胶，然后经结晶致孔法制备了内嵌纳凝胶的甲基丙烯酸羟乙酯纳晶胶，对其接枝功能单体N，N，N-三甲基乙烯基苯甲氯化铵，得到内嵌纳凝胶阴离子交换纳晶胶，用于从发酵液中分离苯乳酸。结果表明：所得纳晶胶具有良好的渗透性能和轴向分散性能，在流速为1 cm·min^-1时，不同配比纳晶胶对牛血清白蛋白的吸附容量为3.9~5.4 mg·ml^-1；将质量比为 8∶2（HEMA∶pBMA）的纳晶胶用于从转化液中分离苯乳酸，所得苯乳酸的纯度达89.24%，回收率93.74%，分离效果好，表明其在生物分离方面有良好的应用前景。

关键词: 纳凝胶, 纳晶胶, 吸附, 苯乳酸

CLC Number:

TQ 028

HE Yawei,ZHANG Songhong,HUANG Jie,LIU Liu,LI Guohua,YUN Junxian. Separation of phenyllactic acid from transformation broth by anion exchange poly(2-hydroxyethyl methacrylate) composite cryogel embedded with nanogels[J]. CIESC Journal, 2020, 71(12): 5636-5643.

贺亚维,张颂红,黄杰,刘流,李国华,贠军贤. 内嵌纳凝胶阴离子交换聚甲基丙烯酸羟乙酯复合晶胶分离苯乳酸研究[J]. 化工学报, 2020, 71(12): 5636-5643.

Figures/Tables 11

Fig.1 FT-IR spectra of BMA, EDMA, pBMA

Fig.2 Particle size distribution of pBMA nanogels

Fig.3 Scanning electron microscope photographs of anion-exchange nano-cryogels

Table 1 The porosities of nano-cryogels

Mass ratios of HEMA to pBMA nanogels	The effective porosity/%	The absolute porosity /%
9:1	82.5	88.0
8:2	81.2	88.8
7:3	79.5	84.9

Table 2 Permeability and length of the composite cryogel matrix with different mass ratios of HEMA to pBMA nanogels (column inner diameter: 10 mm)

Mass ratios of HEMA to pBMA nanogels	L/cm	k_w×10¹²/m²
9:1	5.0	1.77
8:2	4.7	1.26
7:3	5.2	0.35

Fig.4 Residence time distribution curves in nano-cryogels

Fig.5 HETP values of nano-cryogels at various liquid velocities

Fig.6 Chromatographic curves of BSA in the nano-cryogels with different mass ratios of HEMA to pBMA nanogels

Fig.7 Chromatographic adsorption process of PLA in the nano-cryogel

Fig.8 Concentrations and purities of PLA with the loading liquid volume during the chromatography process of sample solution

Fig.9 HPLC results of PLA and other contamination contents in the effluent sample for the chromatography process of sample solution

References 42

1	Dieuleveux V, van der Pyl D, Chataud J, et al. Purification and characterization of anti-Listeria compounds produced by Geotrichum candidum[J]. Applied Environmental Microbiology, 1998, 64(2): 800-803.
2	Schwenninger S M, Lacroix C, Truttmann S, et al. Characterization of low-molecular-weight antiyeast metabolites produced by a food-protective Lactobacillus-Propionibacterium coculture[J]. Journal of Food Protection, 2008, 71(12): 2481-2487.
3	Prema P, Smila D, Palavesam A, et al. Production and characterization of an antifungal compound (3-phenyllactic acid) produced by Lactobacillus plantarum strain[J]. Food and Bioprocess Technology, 2010, 3(3): 379-386.
4	Mu W M, Yu S H, Zhu L J, et al. Recent research on 3-phenyllactic acid, a broad-spectrum antimicrobial compound[J]. Applied Microbiology Biotechnology, 2012, 95(5): 1155-1163.
5	Li L, Shin S Y, Lee K W, et al. Production of natural antimicrobial compound D-phenyllactic acid using Leuconostoc mesenteroides ATCC8293 whole cells involving highly active D-lactate dehydrogenase[J]. Letters Applied Microbiology, 2014, 59(4): 404-411.
6	Ning Y W, Yan A H, Yang K, et al. Antibacterial activity of phenyllactic acid against Listeria monocytogenes and Escherichia coli by dual mechanisms[J]. Food Chemistry, 2017, 228: 533-540.
7	Fujita T, Nguyen H D, Ito T, et al. Microbial monomers custom-synthesized to build true bio-derived aromatic polymers[J]. Applied Microbiology and Biotechnology, 2013, 97(20): 8887-8894.
8	Valerio F, Di Blase M, Lattanzio V M, et al. Improvement of the antifungal activity of lactic acid bacteria by addition to the growth medium of phenylpyruvic acid, a precursor of PLA[J]. International Journal of Food Microbiology, 2016, 222: 1-7.
9	朱银龙, 贠军贤, 沈绍传, 等. 透性化干酪乳杆菌细胞转化苯丙酮酸合成苯乳酸[J]. 高校化学工程学报, 2015, 29(2): 495-500.
	Zhu Y L, Yun J X, Shen S C, et al. Biotransformation of phenylpyruvic acid into phenyllactic acid with permeabilized Lactobacillus casei cells [J]. Journal of Chemical Engineering of Chinese Universities, 2015, 29(2): 495-500.
10	Rodríguez N, Salgado J M, Cortés S, et al. Antimicrobial activity of D-3-phenyllactic acid produced by fed-batch process against Salmonella enteric[J]. Food Control, 2012, 25(1): 274-284.
11	Li X F, Jiang B, Pan B L. Biotransformation of phenylpyruvic acid to phenyllactic acid by growing and resting cells of a Lactobacillus sp[J]. Biotechnology Letters, 2007, 29(4): 593-597.
12	倪正, 关今韬, 沈绍传, 等. 苯乳酸的微生物合成及分离研究进展[J]. 化工进展, 2016, 35(11): 3627-3633.
	Ni Z, Guan J T, Shen S C. An overview of recent advances in microbial synthesis and separation of phenyllactic acid[J]. Chemical Industry and Engineering Progress, 2016, 35(11): 3627-3633.
13	Gizem E, Bo M. Cryogels-versatile tools in bioseparation[J]. Journal of Chromatography A, 2014, 1357(8): 24-35.
14	Lozinsky V I, Galaev I Y, Plieva F M, et al. Polymeric cryogels as promising materials of biotechnological interest[J]. Trends in Biotechnology, 2003, 21(10): 445-451.
15	Plieva F M , Galaev I Y, Mattlasson B. Macroporous gels prepared at subzero temperatures as novel materials for chromatography of particulate-containing fluids and cell culture applications[J]. Journal of Separation Science, 2007, 30(11): 1657-1671.
16	Plieva F M, Kirsebom H, Mattlasson B. Preparation of macroporous cryostructurated gel monoliths, their characterization and main applications[J]. Journal of Separation Science, 2011, 34(16/17): 2164-2172.
17	Yao K J, Shen S C, Yun J X, et al. Preparation of polyacrylamide-based supermacroporous monolithic cryogelbeds under freezing-temperature variation conditions[J]. Chemical Engineering Science, 2006, 61(20): 6701-6708.
18	Hanora A, Plieva F M, Hedström M, et al. Capture of bacterial endotoxins using a supermacroporous monolithic matrix with immobilized polyethyleneimine, lysozyme or polymyxin B[J]. Journal of Biotechnology, 2005, 118(4): 421-433.
19	Arvidsson P, Pleeva F M, Lozinsky V I, et al. Direct chromatographic capture of enzyme from crude homogenate using immobilized metal affinity chromatography on a continuous supermacroporous adsorbent[J]. Journal of Chromatography A, 2003, 986(2): 275-290.
20	Arvidsson P, Pleeva F M, Savina I N, et al. Chromatography of microbial cells using continuous supermacroporous affinity and ion-exchange columns[J]. Journal of Chromatography A, 2002, 977(1): 27-38.
21	Tekin K, Uzun L, Şahin Ç, et al. Preparation and characterization of composite cryogels containing imidazole group and use in heavy metal removal[J]. Reactive and Functional Polymers, 2011, 71(10): 985-993.
22	Yao K J, Yun J X, Shen S C, et al. Characterization of a novel continuous supermacroporous monolithic cryogel embedded with nanoparticles for protein chromatography[J]. Journal of Chromatography A, 2006, 1109(1): 103-110.
23	Bruve L J, Chase H A. Hydrodynamics and adsorption behavior within an expanded bed adsorption column studied using in-bed sampling[J]. Chemical Engineering Science, 2011, 56(10): 3149-3162.
24	Uygun M, Akduman B, Akgöl S, et al. A new metal-chelated cryogel for reversible immobilization of urease[J]. Applied Biochemistry and Biotechnology, 2013, 170(8): 1815-1826.
25	Savina I N, Galaev I Y, Mattiasson B.Anion-exchange supermacroporous monolithic matrices with grafted polymer brushes of N, N-dimethylaminoethyl-methacrylate[J]. Journal of Chromatography A, 2005, 1092(2): 199-205.
26	Yun J X, Cheng X H, Ye J L, et al. Chromatographic adsorption of serum albumin and antibody proteins in cryogels with benzyl-quaternary amine ligands[J]. Journal of Chromatography A, 2015, 1381: 173-183.
27	Ye J L, Yun J X, Lin D Q, et al. Poly(hydroxyethyl methacrylate)-based composite cryogel with embedded macroporous cellulose beads for the separation of human serum immunoglobulin and albumin[J]. Journal of Separation Science, 2013, 36: 3813-3820.
28	Pan M, Shen S, Chen L, et al. Separation of lactoperoxidase from bovine whey milk by cation exchange composite cryogel embedded macroporous cellulose beads[J]. Separation & Purification Technology, 2015, 147: 132-138.
29	Kangkamano T, Numnuam A, Limbut W, et al. Chitosan cryogel with embedded gold nanoparticles decorated multiwalled carbon nanotubes modified electrode for highly sensitive flow based non-enzymatic glucose sensor[J]. Sensors & Actuators B Chemical, 2017, 246: 854-863.
30	Tao S P, Wang C, Sun Y. Coating of nanoparticles on cryogel surface and subsequent double-modification for enhanced ion-exchange capacity of protein[J]. Journal of Chromatography A, 2014, 1359: 76-83.
31	Alkan H, Cömert Ş C, Gürbüz F, et al. Cu2+-attached pumice particles embedded composite cryogels for protein purification[J]. Artificial Cells, Nanomedicine, and Biotechnology, 2017, 45(1): 90-97.
32	Wang C, Dong X Y, Jiang Z, et al. Enhanced adsorption capacity of cryogel bed by incorporating polymeric resin particles[J]. Journal of Chromatography A, 2013, 1272 (11): 20-25.
33	刘杰, 沈绍传, 陈平, 等. 内嵌纳米粒阴离子交换聚甲基丙烯酸羟乙酯复合晶胶分离三磷酸胞苷[J]. 化工学报, 2014, 65(10): 3938-3945.
	Liu J, Shen S C, Chen P, et al. Separation of cytidine triphosphate from Saccharomyces cerevisiae broth by anion exchange poly(2-hydroxyethyl methacrylate) composite cryogel embedded with SiO2 nanoparticles[J]. CIESC Journal, 2014, 65(10): 3938-3945.
34	Ma Y K, Ge Y X, Li L B. Advancement of multifunctional hybrid nanogel systems: construction and application in drug co-delivery and imaging technique[J]. Materials Science and Engineering C, 2017, 71(2): 1281-1292.
35	Oh J K, Lee D I, Park J M. Biopolymer-based microgels/nanogels for drug delivery applications[J]. Progress in Polymer Science, 2009, 34 (8): 1261-1282.
36	Oh J K, Drumright R, Siegwart D J, et al. The development of microgels/nanogels for drug delivery applications[J]. Progress in Polymer Science, 2008, 33(4): 448-477.
37	Sasaki Y, Akiyoshi K. Nanogel engineering for new nanobiomaterials: from chaperoning engineering to biomedical applications[J]. Chemical Record, 2010, 10(6): 366-376.
38	Soni G, Yadav K S. Nanogels as potential nanomedicine carrier for treatment of cancer: a mini review of the state of the art[J]. Saudi Pharmaceutical Journal, 2016, 24(2): 133-139.
39	Wu H Q, Wang C C. Biodegradable smart nanogels: a new platform for targeting drug delivery and biomedical diagnostics[J]. Langmuir, 2016, 32(25): 6211-6225.
40	Branniguan R P, Khutoryanskiyv V. Synthesis and evaluation of mucoadhesive acryloyl-quaternized PDMAEMA nanogels for ocular drug delivery[J]. Colloids and Surfaces B: Biointerfaces, 2017, 155: 538-543.
41	Ahmed I N, Chang R, Tsai W B. Poly(acrylic acid) nanogel as a substrate for cellulase immobilization for hydrolysis of cellulose[J]. Colloids and Surfaces B: Biointerfaces, 2017, 152: 339-343.
42	Guan J T, Guan Y X, Yun J X, et al. Chromatographic separation of phenyllactic acid from crude broth using cryogels with dual functional groups[J]. Journal of Chromatography A, 2018, 1554(11): 92-100.

Related Articles 15

[1]	Bingchun SHENG, Jianguo YU, Sen LIN. Study on lithium resource separation from underground brine with high concentration of sodium by aluminum-based lithium adsorbent [J]. CIESC Journal, 2023, 74(8): 3375-3385.
[2]	Ruihang ZHANG, Pan CAO, Feng YANG, Kun LI, Peng XIAO, Chun DENG, Bei LIU, Changyu SUN, Guangjin CHEN. Analysis of key parameters affecting product purity of natural gas ethane recovery process via ZIF-8 nanofluid [J]. CIESC Journal, 2023, 74(8): 3386-3393.
[3]	Yan GAO, Peng WU, Chao SHANG, Zejun HU, Xiaodong CHEN. Preparation of magnetic agarose microspheres based on a two-fluid nozzle and their protein adsorption properties [J]. CIESC Journal, 2023, 74(8): 3457-3471.
[4]	Ji CHEN, Ze HONG, Zhao LEI, Qiang LING, Zhigang ZHAO, Chenhui PENG, Ping CUI. Study on coke dissolution loss reaction and its mechanism based on molecular dynamics simulations [J]. CIESC Journal, 2023, 74(7): 2935-2946.
[5]	Jie WANG, Xiaolin QIU, Ye ZHAO, Xinyang LIU, Zhongqiang HAN, Yong XU, Wenhan JIANG. Preparation and properties of polyelectrolyte electrostatic deposition modified PHBV antioxidant films [J]. CIESC Journal, 2023, 74(7): 3068-3078.
[6]	Caihong LIN, Li WANG, Yu WU, Peng LIU, Jiangfeng YANG, Jinping LI. Effect of alkali cations in zeolites on adsorption and separation of CO₂/N₂O [J]. CIESC Journal, 2023, 74(5): 2013-2021.
[7]	Chenxin LI, Yanqiu PAN, Liu HE, Yabin NIU, Lu YU. Carbon membrane model based on carbon microcrystal structure and its gas separation simulation [J]. CIESC Journal, 2023, 74(5): 2057-2066.
[8]	Shaoyun CHEN, Dong XU, Long CHEN, Yu ZHANG, Yuanfang ZHANG, Qingliang YOU, Chenglong HU, Jian CHEN. Preparation and adsorption properties of monolayer polyaniline microsphere arrays [J]. CIESC Journal, 2023, 74(5): 2228-2238.
[9]	Yu PAN, Zihang WANG, Jiayun WANG, Ruzhu WANG, Hua ZHANG. Heat and moisture performance study of Cur-LiCl coated heat exchanger [J]. CIESC Journal, 2023, 74(3): 1352-1359.
[10]	Xuanjun WU, Chao WANG, Zijian CAO, Weiquan CAI. Deep learning model of fixed bed adsorption breakthrough curve hybrid-driven by data and physical information [J]. CIESC Journal, 2023, 74(3): 1145-1160.
[11]	Xiaowan PENG, Xiaonan GUO, Chun DENG, Bei LIU, Changyu SUN, Guangjin CHEN. Modeling and simulation of CH₄/N₂ separation process with two absorption-adsorption columns using ZIF-8 slurry [J]. CIESC Journal, 2023, 74(2): 784-795.
[12]	Jinlin MENG, Yu WANG, Qunfeng ZHANG, Guanghua YE, Xinggui ZHOU. Pore network model of low-temperature nitrogen adsorption-desorption in mesoporous materials [J]. CIESC Journal, 2023, 74(2): 893-903.
[13]	Wan XU, Zhenbin CHEN, Huijuan ZHANG, Fangfang NIU, Ting HUO, Xingsheng LIU. Study on synthesis, adsorption and desorption performance of linear temperature-sensitive segment polymer regulated intelligent $R e O 4 -$ ion-imprinted polymer [J]. CIESC Journal, 2023, 74(2): 941-952.
[14]	Jiahao JIANG, Xiaole HUANG, Jiyun REN, Zhengrong ZHU, Lei DENG, Defu CHE. Qualitative and quantitative study on Pb²⁺ adsorption by biochar in solution [J]. CIESC Journal, 2023, 74(2): 830-842.
[15]	Yingxi DANG, Peng TAN, Xiaoqin LIU, Linbing SUN. Temperature swing for CO₂ capture driven by radiative cooling and solar heating [J]. CIESC Journal, 2023, 74(1): 469-478.