改性聚乙烯膜和氧化硅固定化碳酸酐酶

doi:10.11949/0438-1157.20230348

摘要/Abstract

摘要：

分别以改性聚乙烯（PE）膜和氧化硅（SiO₂）为载体，考察了不同方法固定化碳酸酐酶（CA）的活性，然后以聚多巴胺/聚乙烯亚胺（PDA/PEI）改性的PE和SiO₂固定化CA为研究对象，考察了最适反应条件及稳定性。结果表明：在相同的反应条件下，PDA/PEI-SiO₂固定化CA的酶活回收率最高，为58.8%，PDA/PEI-PE固定化CA的酶活回收率为17.1%，使用10次后保留活性分别为84.8%和90.2%；固定化酶的最适反应条件均为35℃、pH 8.5，与游离酶相同；在较高温度（55~65℃）和较高酸浓度（>0.010 mol/L）下的稳定性高于游离酶；Mn²⁺对游离酶和固定化酶活性均有显著的促进作用，K⁺、Mg²⁺无明显影响；PDA/PEI-SiO₂和PDA/PEI-PE固定化CA在4℃下贮存10 d，保留活性分别为95.2%和92.4%；当用于CO₂水合反应时，CaCO₃的生成量分别是游离CA的120%和70%。固定化CA在CO₂捕集的工业化应用方面具有很大潜力。

关键词: 碳酸酐酶, 固定化, 聚乙烯, 氧化硅, 稳定性, 酶活

Abstract:

The activity of carbonic anhydrase (CA) immobilized by different methods was investigated using modified polyethylene (PE) membrane and silica (SiO₂) as carriers, and then modified with polydopamine/polyethyleneimine (PDA/PEI) PE- and SiO₂-immobilized CA were used as research objects. The optimum reaction conditions and stability were investigated. The results show that under the same reaction conditions, the activity recovery of PDA/PEI-SiO₂ immobilized CA was the highest, which was 58.8%, and the activity recovery of PDA/PEI-PE immobilized CA was 17.1%. Their retentive activities were 84.8% and 90.2%, respectively, after 10 use recycles. The optimum reaction conditions of immobilized enzymes were 35℃ and pH 8.5, which were the same as those of free enzymes. The stability of the two immobilized enzymes at higher temperature (55—65℃) and higher acid concentration (>0.010 mol/L) was better than that of free enzyme. Mg²⁺ could significantly promote the activity of free enzyme and immobilized enzyme, while K⁺ and Mg²⁺ had no obvious effect. PDA/PEI-SiO₂ and PDA/PEI-PE immobilized CA retained 95.2%和92.4% activity after stored at 4℃ for 10 d. When used in the CO₂ hydration reaction, the amount of CaCO₃ produced was 120% and 70% of free CA. The immobilized CA has great potential in the industrial application of CO₂ capture.

Key words: carbonic anhydrase, immobilization, polyethylene, silica, stability, enzyme activity

中图分类号:

TQ 814.2

孟令玎, 崇汝青, 孙菲雪, 孟子晖, 刘文芳. 改性聚乙烯膜和氧化硅固定化碳酸酐酶[J]. 化工学报, 2023, 74(8): 3472-3484.

Lingding MENG, Ruqing CHONG, Feixue SUN, Zihui MENG, Wenfang LIU. Immobilization of carbonic anhydrase on modified polyethylene membrane and silica[J]. CIESC Journal, 2023, 74(8): 3472-3484.

图/表 12

图1 CA在不同载体上的固定化路线

Fig.1 Immobilization routes of CA on different supports

图2 改性前后PE膜的SEM图

Fig.2 SEM images of PE films before and after modification

图3 改性前后SiO2的SEM图

Fig.3 SEM images of SO2 before and after modification

图4 载体改性前后的FTIR谱图

Fig.4 FTIR spectra of carriers before and after modification

表1 SiO2改性前后的表面Zeta电位

Table 1 Surface Zeta potential of SiO2 before and after modification

Samples	Zeta potential/mV
SiO₂	-51.5
PDA/PEI-SiO₂	+71.6
KH550-SiO₂	+20.5
KH560-SiO₂	-24.1

图5 不同载体和路线固定化CA的催化性能

Fig.5 Catalytic performance of CA immobilized by different carriers and different routes

图6 反应温度和pH对固定化CA活性的影响

Fig.6 Effects of reaction temperature and pH on the activity of immobilized CA

图7 固定化CA的热稳定性

Fig.7 Thermal stability of immobilized CA

图8 固定化CA的耐酸性

Fig.8 Acid resistance of immobilized CA

图9 固定化CA的离子稳定性

Fig.9 Ions stability of immobilized CA

图10 固定化CA的贮存稳定性

Fig.10 Storage stability of immobilized CA

表2 固定化CA的固碳性能

Table 2 CO2 sequestration performance of immobilized CA

Supports	$m_{C a C O_{3}}$ /mg	m_CA/mg	( $m_{C a C O_{3}}$ /m_CA)/ (mg/mg)	Ref.
Free CA	12.84	0.2	64.2	this work
PDA/PEI-SiO₂	15.68	0.2	78.4	this work
PDA/PEI-PE	9.33	0.2	46.6	this work
Mesoporous aluminosilicate	16.14	1	16.1	[51]
Alumino-siloxane aerogel beads	12	0.017	705.9	[52]
KH550-PVDF	~8	0.8	10.0	[13]
KH560-PVDF	~6	0.8	7.5	[13]

表2 固定化CA的固碳性能

Table 2 CO2 sequestration performance of immobilized CA

Supports	$m_{C a C O_{3}}$ /mg	m_CA/mg	( $m_{C a C O_{3}}$ /m_CA)/ (mg/mg)	Ref.
Free CA	12.84	0.2	64.2	this work
PDA/PEI-SiO₂	15.68	0.2	78.4	this work
PDA/PEI-PE	9.33	0.2	46.6	this work
Mesoporous aluminosilicate	16.14	1	16.1	[51]
Alumino-siloxane aerogel beads	12	0.017	705.9	[52]
KH550-PVDF	~8	0.8	10.0	[13]
KH560-PVDF	~6	0.8	7.5	[13]

参考文献 52

1	Talekar S, Jo B H, Dordick J S, et al. Carbonic anhydrase for CO₂capture, conversion and utilization[J]. Current Opinion in Biotechnology, 2022, 74: 230-240.
2	Asif M, Suleman M, Haq I, et al. Post-combustion CO₂capture with chemical absorption and hybrid system: current status and challenges[J]. Greenhouse Gases: Science and Technology, 2018, 8(6): 998-1031.
3	Saeed I M, Alaba P, Mazari S A, et al. Opportunities and challenges in the development of monoethanolamine and its blends for post-combustion CO₂ capture[J]. International Journal of Greenhouse Gas Control, 2018, 79: 212-233.
4	Bernhardsen I M, Knuutila H K. A review of potential amine solvents for CO₂ absorption process: absorption capacity, cyclic capacity and pK_a [J]. International Journal of Greenhouse Gas Control, 2017, 61: 27-48.
5	Aghel B, Janati S, Wongwises S, et al. Review on CO₂ capture by blended amine solutions[J]. International Journal of Greenhouse Gas Control, 2022, 119: 103715.
6	梁珊, 宗敏华, 娄文勇. 酶法催化二氧化碳制备高附加值化学品研究进展[J]. 化学学报, 2019, 77(11): 1099-1114.
	Liang S, Zong M H, Lou W Y. Recent advances in enzymatic catalysis for preparation of high value-added chemicals from carbon dioxide[J]. Acta Chimica Sinica, 2019, 77(11): 1099-1114.
7	Sun M K, Alkon D L. Carbonic anhydrase gating of attention: memory therapy and enhancement[J]. Trends in Pharmacological Sciences, 2002, 23(2): 83-89.
8	Sharma T, Sharma S, Kamyab H, et al. Energizing the CO₂ utilization by chemo-enzymatic approaches and potentiality of carbonic anhydrases: a review[J]. Journal of Cleaner Production, 2020, 247: 119138.
9	孟令玎, 毛梦雷, 廖奇勇等. 碳酸酐酶和甲酸脱氢酶的稳定性研究进展[J]. 化工进展, 2022, 41(S1): 436-447.
	Meng L D, Mao M L, Liao Q Y, et al. Recent advance in stability of carbonic anhydrase and formate dehydrogenase[J]. Chemical Industry and Engineering Progress, 2022, 41(S1): 436-447.
10	De Oliveira Maciel A, Christakopoulos P, Rova U, et al. Carbonic anhydrase to boost CO₂ sequestration: improving carbon capture utilization and storage (CCUS)[J]. Chemosphere, 2022, 299: 134419.
11	Molina-Fernández C, Luis P. Immobilization of carbonic anhydrase for CO₂ capture and its industrial implementation: a review[J]. Journal of CO₂ Utilization, 2021, 47: 101475.
12	Rasouli H, Iliuta I, Bougie F, et al. Enzyme-immobilized flat-sheet membrane contactor for green carbon capture[J]. Chemical Engineering Journal, 2021, 421: 129587.
13	Sun J, Wei L, Wang Y, et al. Immobilization of carbonic anhydrase on polyvinylidene fluoride membranes[J]. Biotechnology and Applied Biochemistry, 2018, 65(3): 362-371.
14	Shen J, Yuan Y, Salmon S. Carbonic anhydrase immobilized on textile structured packing using chitosan entrapment for CO₂ capture[J]. ACS Sustainable Chemistry & Engineering, 2022, 10(23): 7772-7785.
15	Rasouli H, Iliuta I, Bougie F, et al. Enhanced CO₂ capture in packed-bed column bioreactors with immobilized carbonic anhydrase[J]. Chemical Engineering Journal, 2022, 432: 134029.
16	Chang S, He Y, Li Y, et al. Study on the immobilization of carbonic anhydrases on geopolymer microspheres for CO₂ capture[J]. Journal of Cleaner Production, 2021, 316: 128163.
17	Fei X, Chen S, Liu D, et al. Comparison of amino and epoxy functionalized SBA-15 used for carbonic anhydrase immobilization[J]. Journal of Bioscience and Bioengineering, 2016, 122(3): 314-321.
18	Xu J, Zhang Z, Pang S, et al. Accelerated CO₂ capture using immobilized carbonic anhydrase on polyethyleneimine/dopamine co-deposited MOFs[J]. Biochemical Engineering Journal, 2022, 189: 108719.
19	Wang Y, Chen Y, Wang C, et al. Polyethylenimine-modified membranes for CO₂ capture and in situ hydrogenation[J]. ACS Applied Materials & Interfaces, 2018, 10(34): 29003-29009.
20	Sun J, Wang C, Wang Y, et al. Immobilization of carbonic anhydrase on polyethylenimine/dopamine codeposited membranes[J]. Journal of Applied Polymer Science, 2019, 136(29): 47784.
21	王彩红, 孙婧, 季书馨等. 聚乙烯亚胺/多巴胺改性氧化硅固定碳酸酐酶[J]. 化工学报, 2019, 70(5): 1887-1893.
	Wang C H, Sun J, Ji S X, et al. Immobilization of carbonic anhydrase on polyethylenimine/dopamine co-deposited SiO₂ [J]. CIESC Journal, 2019, 70(5): 1887-1893.
22	Zhang Q, Huang R, Guo L H. One-step and high-density protein immobilization on epoxysilane-modified silica nanoparticles[J]. Chinese Science Bulletin, 2009, 54(15): 2620-2626.
23	Ozdemir E. Biomimetic CO₂ sequestration: 1. Immobilization of carbonic anhydrase within polyurethane foam[J]. Energy & Fuels, 2009, 23(11): 5725-5730.
24	Yang H C, Liao K J, Huang H, et al. Mussel-inspired modification of a polymer membrane for ultra-high water permeability and oil-in-water emulsion separation[J]. Journal of Materials Chemistry A, 2014, 2(26): 10225-10230.
25	Lü Y, Lu G, Wang Y, et al. Functionalization of cubic Ia3d mesoporous silica for immobilization of penicillin G acylase[J]. Advanced Functional Materials, 2007, 17(13): 2160-2166.
26	Bamane P B, Jagtap R N. Synthesis of the hydrophilic additive by grafting glycidyloxypropyl trimethoxysilane on hydrophilic nanosilica and its modification by using dimethyl propionic acid for self-cleaning coatings[J]. Colloid and Interface Science Communications, 2021, 43: 100444.
27	Hou J, Ji C, Dong G, et al. Biocatalytic Janus membranes for CO₂ removal utilizing carbonic anhydrase[J]. Journal of Materials Chemistry A, 2015, 3(33): 17032-17041.
28	Jun S H, Yang J, Jeon H, et al. Stabilized and immobilized carbonic anhydrase on electrospun nanofibers for enzymatic CO₂ conversion and utilization in expedited microalgal growth[J]. Environmental Science & Technology, 2020, 54(2): 1223-1231.
29	Lee H, Dellatore Shara M, Miller William M, et al. Mussel-inspired surface chemistry for multifunctional coatings[J]. Science, 2007, 318(5849): 426-430.
30	Dai M, Huang T, Chao L, et al. Horseradish peroxidase-catalyzed polymerization of l-DOPA for mono-/bi-enzyme immobilization and amperometric biosensing of H₂O₂and uric acid[J]. Talanta, 2016, 149: 117-123.
31	Hou J, Dong G, Xiao B, et al. Preparation of titania based biocatalytic nanoparticles and membranes for CO₂conversion[J]. Journal of Materials Chemistry A, 2015, 3(7): 3332-3342.
32	Shao P, Chen H, Ying Q, et al. Structure–activity relationship of carbonic anhydrase enzyme immobilized on various silica-based mesoporous molecular sieves for CO₂ absorption into a potassium carbonate solution[J]. Energy & Fuels, 2020, 34(2): 2089-2096.
33	Cantone S, Ferrario V, Corici L, et al. Efficient immobilisation of industrial biocatalysts: criteria and constraints for the selection of organic polymeric carriers and immobilisation methods[J]. Chemical Society Reviews, 2013, 42(15): 6262-6276.
34	Jing G, Pan F, Lv B, et al. Immobilization of carbonic anhydrase on epoxy-functionalized magnetic polymer microspheres for CO₂ capture[J]. Process Biochemistry, 2015, 50(12): 2234-2241.
35	Wanjari S, Prabhu C, Yadav R, et al. Immobilization of carbonic anhydrase on chitosan beads for enhanced carbonation reaction[J]. Process Biochemistry, 2011, 46(4): 1010-1018.
36	Lavecchia R, Zugaro M. Thermal denaturation of erythrocyte carbonic anhydrase[J]. FEBS Letters, 1991, 292(1): 162-164.
37	Sarraf N S, Saboury A A, Ranjbar B, et al. Structural and functional changes of bovine carbonic anhydrase as a consequence of temperature[J]. Acta Biochimica Polonica, 2004, 51(3): 665-671.
38	Pocker Y, Stone J T. The catalytic versatility of erythrocyte carbonic anhydrase(Ⅲ): Kinetic studies of the enzyme-catalyzed hydrolysis of p-nitrophenyl acetate[J]. Biochemistry, 1967, 6(3): 668-678.
39	Reetz Manfred T, Sun Z, Qu G. Enzyme Engineering: Selective Catalysts for Applications in Biotechnology, Organic Chemistry, and Life Science [M]. Wiley-VCH GmbH, 2023.
40	李宇彤, 白姝, 余林玲. PEI接枝纳米二氧化硅载体的构建及碳酸酐酶的固定化[J]. 离子交换与吸附, 2019, 35(4): 300-311.
	Li Y T, Bai S, Yu L L. Synthesis of polyethyleneimine-grafted silica nanoparticles and carbonic anhydrase immobilization[J]. Ion Exchange and Adsorption, 2019, 35(4): 300-311.
41	Zhu Y, Li W, Sun G, et al. Enzymatic properties of immobilized carbonic anhydrase and the biocatalyst for promoting CO₂ capture in vertical reactor[J]. International Journal of Greenhouse Gas Control, 2016, 49: 290-296.
42	Mao M, Zhai T, Meng L, et al. Controllable preparation of mesoporous silica and its application in enzyme-catalyzed CO₂ reduction[J]. Chemical Engineering Journal, 2022, 437: 135479.
43	Ren S, Li C, Tan Z, et al. Carbonic anhydrase@ZIF-8 hydrogel composite membrane with improved recycling and stability for efficient CO₂ capture[J]. Journal of Agricultural and Food Chemistry, 2019, 67(12): 3372-3379.
44	Gitlin I, Carbeck J D, Whitesides G M. Why are proteins charged? Networks of charge-charge interactions in proteins measured by charge ladders and capillary electrophoresis[J]. Angewandte Chemie International Edition, 2006, 45(19): 3022-3060.
45	Schofield K. Mercury emission control from coal combustion systems: a modified air preheater solution[J]. Combustion and Flame, 2012, 159(4): 1741-1747.
46	朱轶林. 固定化碳酸酐酶的酶学性质和催化吸收CO₂特性的实验研究[D]. 重庆: 重庆大学, 2015.
	Zhu Y L. Research on enzymatic proprities of immobilized carbonic anhydrase and CO₂ catalytic absorption[D]. Chongqing: Chongqing University, 2015.
47	Wu Y, Zhao X, Li P, et al. Impact of Zn, Cu, and Fe on the activity of carbonic anhydrase of erythrocytes in ducks[J]. Biological Trace Element Research, 2007, 118(3): 227-232.
48	Tan S I, Han Y L, Yu Y J, et al. Efficient carbon dioxide sequestration by using recombinant carbonic anhydrase[J]. Process Biochemistry, 2018, 73: 38-46.
49	Ramanan R, Kannan K, Sivanesan S D, et al. Bio-sequestration of carbon dioxide using carbonic anhydrase enzyme purified from Citrobacter freundii [J]. World Journal of Microbiology and Biotechnology, 2009, 25(6): 981-987.
50	Kim Y K, Lee S Y, Oh B K. Enhancement of formic acid production from CO₂ in formate dehydrogenase reaction using nanoparticles[J]. RSC Advances, 2016, 6(111): 109978-109982.
51	Wanjari S, Prabhu C, Satyanarayana T, et al. Immobilization of carbonic anhydrase on mesoporous aluminosilicate for carbonation reaction[J]. Microporous and Mesoporous Materials, 2012, 160: 151-158.
52	Shamna I, Kwan Jeong S, Margandan B. Covalent immobilization of carbonic anhydrase on amine functionalized alumino-siloxane aerogel beads for biomimetic sequestration of CO₂ [J]. Journal of Industrial and Engineering Chemistry, 2021, 100: 288-295.

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