酶自固定化方法研究进展

doi:10.16085/j.issn.1000-6613.2018-0605

摘要/Abstract

摘要： 固定化酶有诸多优势，传统酶固定化方法通常需要外界辅助，但额外的固定化流程无疑会增加酶使用成本。近年来提出的酶自固定化则可在温和条件下实现自身固定化，简化固定流程，为降低酶的成本、促进酶的广泛应用提供了新思路。本文系统、深入介绍了无载体类型、自催化类型和自结合类型酶自固定化的方法，并指出其各自特点：无载体类型无需载体成本；自催化类型能有效降低载体成本，获得机械强度较高的固定化酶；自结合类型可在温和条件下易实现多酶固定。结合本文作者课题组的相关研究，阐述了酶自身催化的自固定化和蛋白或多肽介导的融合酶自固定化最新研究进展。最后指出当前研究的重点应致力于自固定化新方法的研发，为后期理性选择、合理组合自固定化方法奠定基础，以加快自固定化酶实际应用。

关键词: 固定化酶, 自固定化, 自组装, 自催化, 无载体固定化

Abstract: Immobilized enzymes offer many advantagesbut the traditional immobilization methods usually require external assistance. The additional immobilization process will undoubtedly increase the cost of using enzymes. The self-immobilized enzymes proposed in recent years can perform under mild conditions, which can simplify the immobilization process, and provide new ideas for reducing the cost of enzymes and broaden the application field of enzymes. In this review, many self-immobilization approaches were systematically and thoroughly presented, including carrier-free self-immobilization, the self-catalyzed carrier-based immobilization, and the self-binding immobilization. Different methods have their own characteristics. Carrier-free self-immobilization avoids the cost of the carrier, and the self-catalyzed carrier-based immobilization can effectively circumvent the cost of the carrier and obtain the enzymes with higher mechanical strength. The self-binding immobilization can easily perform multi-enzyme immobilization under a mild condition. Coupling with the current research progresses on the self-immobilization of enzymes, the most recent developments of enzyme self-immobilization were discussed including the enzyme self-catalyzed immobilization and proteins-mediated (or peptides-mediated) self-immobilization. The current research of enzyme self-immobilization should be focus on the research and development of novel methods, which will lay a foundation for later rational selection and rational combination of self-immobilization methods and accelerate the practical application of self-immobilized enzymes.

Key words: enzyme immobilization, self-immobilization, self-assembly, self-catalysis, carrier-free immobilization

中图分类号:

Q814.2

林源清, 李夏兰, 张光亚. 酶自固定化方法研究进展[J]. 化工进展, 2018, 37(12): 4523-4532.

LIN Yuanqing, LI Xialan, ZHANG Guangya. Recent research progress of enzyme self-immobilization methods[J]. Chemical Industry and Engineering Progress, 2018, 37(12): 4523-4532.

参考文献

[1] DICOSIMO R,MCAULIFFE J,POULOSE A J,et al. Industrial use of immobilized enzymes[J]. Chemical Society Reviews,2013, 42(15):6437-6474.
[2] BRADY D, JORDAAN J, SIMPSON C, et al. Spherezymes:a novel structured self-immobilisation enzyme technology[J]. BMC Biotechnology, 2008, 8:8.
[3] BRADY D, JORDAAN J. Advances in enzyme immobilisation[J]. Biotechnology Letters, 2009, 31(11):1639-1650.
[4] 柯彩霞, 范艳利, 苏枫, 等. 酶的固定化技术最新研究进展[J]. 生物工程学报, 2018, 34(2):1-17. KE Caixia, FAN Yanli, SU Feng, et al. Recent advances in enzyme immobilization[J]. Chinese Journal of Biotechnology, 2018, 34(2):1-17.
[5] SHELDON R A, VAN PELT S. Enzyme immobilisation in biocatalysis:why, what and how[J]. Chemical Society Reviews, 2013, 42(15):6223-6235.
[6] ILLANES A, CAUERHFF A, WILSON L, et al. Recent trends in biocatalysis engineering[J]. Bioresource Technology, 2012, 115:48-57.
[7] SHELDON R A. Characteristic features and biotechnological applications of cross-linked enzyme aggregates (CLEAs)[J]. Applied Microbiology and Biotechnology, 2011, 92(3):467-477.
[8] CUI J D, JIA S R. Optimization protocols and improved strategies of cross-linked enzyme aggregates technology:current development and future challenges[J]. Critical Reviews in Biotechnology, 2013, 35(1):15-28.
[9] MOLAWA L, JORDAAN J, LIMSON J, et al. Modification of alcalase SphereZyme^TM by entrapment in LentiKats^® to impart improved particle stability[J]. Biocatalysis and Biotransformation, 2013, 31(2):71-78.
[10] OVEIMAR BARBOSA A C O B, FERNANDEZ-LAFUENTE R C R D. Glutaraldehyde in biocatalysts design a useful crosslinker and a versatile tool in enzyme immobilization[J]. RSC Advances, 2014, 4(4):1583-1600.
[11] DITZLER L R, SEN A, GANNON M J, et al. Self-assembled enzymatic monolayer directly bound to a gold surface:activity and molecular recognition force spectroscopy studies[J]. Journal of the American Chemical Society, 2011, 133(34):13284-13287.
[12] DICKERSON M B, SANDHAGE K H, NAIK R R. Protein-and peptide-directed syntheses of inorganic materials[J]. Chemical Reviews, 2008, 108(11):4935-4978.
[13] 王生杰, 蔡庆伟, 杜明轩, 等. 二氧化硅的仿生矿化[J]. 化学进展, 2015, 27(2/3):229-241. WANG Shengjie, CAI Qingwei, DU Mingxuan, et al. Biomimetic mineralization of silica[J]. Progress in Chemistry, 2015, 27(2/3):229-241.
[14] LUCKARIFT H R, SPAIN J C, NAIK R R, et al. Enzyme immobilization in a biomimetic silica support[J]. Nature Biotechnology, 2004, 22(2):211-213.
[15] BETANCOR L, LUCKARIFT H R. Bioinspired enzyme encapsulation for biocatalysis[J]. Trends in Biotechnology, 2008, 26(10):566-572.
[16] LUCKARIFT H R, DICKERSON M B, SANDHAGE K H, et al. Rapid, room-temperature synthesis of antibacterial bionanocomposites of lysozyme with amorphous silica or titania[J]. Small, 2006, 2(5):640-643.
[17] SHIOMI T, TSUNODA T, KAWAI A, et al. Biomimetic synthesis of lysozyme-silica hybrid hollow particles using sonochemical treatment:influence of pH and lysozyme concentration on morphology[J]. Chemistry of Materials, 2007, 19(18):4486-4493.
[18] BASSINDALE A R, TAYLOR P G, ABBATE V, et al. Simple and mild preparation of silica-enzyme composites from silicic acid solution[J]. Journal of Materials Chemistry, 2009, 19(41):7606-7609.
[19] ZHOU L, WANG C, JIANG Y, et al. Immobilization of papain in biosilica matrix and its catalytic property[J]. Chinese Journal of Chemical Engineering, 2013, 21(6):670-675.
[20] YANG Y, WANG G, ZHU G, et al. The effect of amorphous calcium phosphate on protein protection against thermal denaturation[J]. Chemical Communications, 2015, 51(41):8705-8707.
[21] GE J, LEI J, ZARE R N. Protein-inorganic hybrid nanoflowers[J]. Nature Nanotechnology, 2012, 7(7):428-432.
[22] CUI J, JIA S. Organic-inorganic hybrid nanoflowers:a novel host platform for immobilizing biomolecules[J]. Coordination Chemistry Reviews, 2017, 352:249-263.
[23] LIU Y, ZHANG Y, LI X, et al. Self-repairing metal-organic hybrid complexes for reinforcing immobilized chloroperoxidase reusability[J]. Chemical Communications, 2017, 53(22):3216-3219.
[24] PATEL S, OTARI S V, LI J, et al. Synthesis of cross-linked protein-metal hybrid nanoflowers and its application in repeated batch decolorization of synthetic dyes[J]. Journal of Hazardous Materials, 2018, 347:442-450.
[25] CHILKOTI A, MEYER D E. Purification of recombinant proteins by fusion with thermally-responsive polypeptides[J]. Nature Biotechnology, 1999, 17(11):1112-1115.
[26] YEBOAH A, COHEN R I, RABOLLI C, et al. Elastin-like polypeptides:a strategic fusion partner for biologics[J]. Biotechnology and Bioengineering, 2016, 113(8):1617-1627.
[27] 李存存, 张光亚. 酶定向固定化方法及应用的研究进展[J]. 化工进展, 2013, 32(10):2467-2474. LI Cuncun, ZHANG Guangya. Research progress of site-specific immobilization of enzymes and application[J]. Chemical Industry and Engineering Progress, 2013, 32(10):2467-2474.
[28] LI C, ZHANG G. The fusions of elastin-like polypeptides and xylanase self-assembled into insoluble active xylanase particles[J]. Journal of Biotechnology, 2014, 177:60-66.
[29] SHANBHAG B K, LIU B, FU J, et al. Self-assembled enzyme nanoparticles for carbon dioxide capture[J]. Nano Letters, 2016, 16(5):3379-3384.
[30] LUO Q, HOU C, BAI Y, et al. Protein assembly:versatile approaches to construct highly ordered nanostructures[J]. Chemical Reviews, 2016, 116(22):13571-13632.
[31] HAUSER C A E, MAURER-STROH S, MARTINS I C. Amyloid-based nanosensors and nanodevices[J]. Chemical Society Reviews, 2014, 43(15):5326-5345.
[32] WEI G, SU Z, REYNOLDS N P, et al. Self-assembling peptide and protein amyloids:from structure to tailored function in nanotechnology[J]. Chemical Society Reviews, 2017, 46(15):4661-4708.
[33] SAMBASHIVAN S, LIU Y, SAWAYA M R, et al. Amyloid-like fibrils of ribonuclease A with three-dimensional domain-swapped and native-like structure[J]. Nature, 2005, 437(7056):266-269.
[34] GUGLIELMI F, MONTI D M, ARCIELLO A, et al. Enzymatically active fibrils generated by the self-assembly of the ApoA-I fibrillogenic domain functionalized with a catalytic moiety[J]. Biomaterials, 2009, 30(5):829-835.
[35] ZHOU X, SHIMANOVICH U, HERLING T W, et al. Enzymatically active microgels from self-assembling protein nanofibrils for microflow chemistry[J]. ACS Nano, 2015, 9(6):5772-5781.
[36] KNOWLES T P J, OPPENHEIM T W, BUELL A K, et al. Nanostructured films from hierarchical self-assembly of amyloidogenic proteins[J]. Nature Nanotechnology, 2010, 5(3):204-207.
[37] KNOWLES T P, FITZPATRICK A W, MEEHAN S, et al. Role of intermolecular forces in defining material properties of protein nanofibrils[J]. Science, 2007, 318(5858):1900-1903.
[38] HEYMAN A, LEVY I, ALTMAN A, et al. SP1 as a novel scaffold building block for self-assembly nanofabrication of submicron enzymatic structures[J]. Nano Letters, 2007, 7(6):1575-1579.
[39] BANEYX F, MUJACIC M. Recombinant protein folding and misfolding in Escherichia coli[J]. Nature Biotechnology, 2004, 22(11):1399-1408.
[40] RINAS U, GARCIA-FRUITÓS E, CORCHERO J L, et al. Bacterial inclusion bodies:discovering their better half[J]. Trends in Biochemical Sciences, 2017, 42(9):726-737.
[41] PARK S, PARK S, CHOI S. Active inclusion body formation using Paenibacillus polymyxa PoxB as a fusion partner in Escherichia coli[J]. Analytical Biochemistry, 2012, 426(1):63-65.
[42] KRAUSS U, JAGER V D, DIENER M, et al. Catalytically-active inclusion bodies-carrier-free protein immobilizates for application in biotechnology and biomedicine[J]. Journal of Biotechnology, 2017, 258:136-147.
[43] WU W, XING L, ZHOU B, et al. Active protein aggregates induced by terminally attached self-assembling peptide ELK16 in Escherichia coli[J]. Microbial Cell Factories, 2011, 10(1):9.
[44] KROGER N, DEUTZMANN R, SUMPER M. Polycationic peptides from diatom biosilica that direct silica nanosphere formation[J]. Science, 1999, 286(5442):1129-1132.
[45] POULSEN N, BERNE C, SPAIN J, et al. Silica immobilization of an enzyme through genetic engineering of the diatom Thalassiosira pseudonana[J]. Angewandte Chemie International Edition, 2007, 46(11):1843-1846.
[46] JO B H, SEO J H, YANG Y J, et al. Bioinspired silica nanocomposite with autoencapsulated carbonic anhydrase as a robust biocatalyst for CO₂ sequestration[J]. ACS Catalysis, 2014, 4(12):4332-4340.
[47] KROGER N, LORENZ S, BRUNNER E, et al. Self-assembly of highly phosphorylated silaffins and their function in biosilica morphogenesis[J]. Science, 2002, 298(5593):584-586.
[48] KROGER N, DEUTZMANN R, SUMPER M. Polycationic peptides from diatom biosilica that direct silica nanosphere formation[J]. Science, 1999, 286(5442):1129-1132.
[49] JIANG Y, SUN Q, JIANG Z, et al. The improved stability of enzyme encapsulated in biomimetic titania particles[J]. Materials Science and Engineering C, 2009, 29(1):328-334.
[50] REN H, ZHANG Y, SU J, et al. Encapsulation of amine dehydrogenase in hybrid titania nanoparticles by polyethylenimine coating and templated biomineralization[J]. Journal of Biotechnology, 2017, 241:33-41.
[51] CARE A, BERGQUIST P L, SUNNA A. Solid-binding peptides:smart tools for nanobiotechnology[J]. Trends in Biotechnology, 2015, 33(5):259-268.
[52] KACAR T, ZIN M T, SO C, et al. Directed self-immobilization of alkaline phosphatase on micro-patterned substrates via genetically fused metal-binding peptide[J]. Biotechnology and Bioengineering, 2009, 103(4):696-705.
[53] YANG M, CHOI B G, PARK T J, et al. Site-specific immobilization of gold binding polypeptide on gold nanoparticle-coated graphene sheet for biosensor application[J]. Nanoscale, 2011, 3(7):2950-2956.
[54] CARE A, NEVALAINEN H, BERGQUIST P L, et al. Effect of Trichoderma reesei proteinases on the affinity of an inorganic-binding peptide[J]. Applied Biochemistry and Biotechnology, 2014, 173(8):2225-2240.
[55] COYLE B L, BANEYX F. A cleavable silica-binding affinity tag for rapid and inexpensive protein purification[J]. Biotechnology and Bioengineering, 2014, 111(10):2019-2026.
[56] COYLE B L, BANEYX F. Direct and reversible immobilization and microcontact printing of functional proteins on glass using a genetically appended silica-binding tag[J]. Chemical Communications, 2016, 52(43):7001-7004.
[57] CHEN X, WANG Y, WANG P. Peptide-induced affinity binding of carbonic anhydrase to carbon nanotubes[J]. Langmuir, 2015, 31(1):397-403.
[58] CARE A, PETROLL K, GIBSON E S Y, et al. Solid-binding peptides for immobilisation of thermostable enzymes to hydrolyse biomass polysaccharides[J]. Biotechnology for Biofuels, 2017, 10(1):29.
[59] OLIVEIRA C, CARVALHO V, DOMINGUES L, et al. Recombinant CBM-fusion technology:applications overview[J]. Biotechnology Advances, 2015, 33(3/4):358-369.
[60] WANG S, CUI G, SONG X, et al. Efficiency and stability enhancement of cis-epoxysuccinic acid hydrolase by fusion with a carbohydrate binding module and immobilization onto cellulose[J]. Applied Biochemistry and Biotechnology, 2012, 168(3):708-717.
[61] KUMAR A, ZHANG S, WU G, et al. Cellulose binding domain assisted immobilization of lipase (GSlip-CBD) onto cellulosic nanogel:characterization and application in organic medium[J]. Colloids and Surfaces B:Biointerfaces, 2015, 136:1042-1050.
[62] LINDER M B. Hydrophobins:proteins that selfassemble at interfaces[J]. Current Opinion in Colloid & Interface Science, 2009, 14(5):356-363.
[63] PISCITELLI A, PENNACCHIO A, LONGOBARDI S, et al. Vmh2 hydrophobin as a tool for the development of "self-immobilizing" enzymes for biosensing[J]. Biotechnology and Bioengineering, 2017, 114(1):46-52.
[64] REHM B H A. Bacterial polymers:biosynthesis, modifications and applications[J]. Nature Reviews Microbiology, 2010, 8(8):578-592.
[65] BLATCHFORD P A, SCOTT C, FRENCH N, et al. Immobilization of organophosphohydrolase OpdA from Agrobacterium radiobacter by overproduction at the surface of polyester inclusions inside engineered Escherichia coli[J]. Biotechnology and Bioengineering, 2012, 109(5):1101-1108.
[66] RAN G, TAN D, DAI W, et al. Immobilization of alkaline polygalacturonate lyase from Bacillus subtilis on the surface of bacterial polyhydroxyalkanoate nano-granules[J]. Applied Microbiology and Biotechnology, 2017, 101(8):3247-3258.
[67] HAY I D, DU J, REYES P R, et al. In vivo polyester immobilized sortase for tagless protein purification[J]. Microbial Cell Factories, 2015, 14(1):190.
[68] MULLANEY J A, REHM B H A. Design of a single-chain multi-enzyme fusion protein establishing the polyhydroxybutyrate biosynthesis pathway[J]. Journal of Biotechnology, 2010, 147(1):31-36.