Green synthesis of enzyme/metal-organic framework composites with high stability in protein denaturing solvents

Abstract

Objectives

Enzyme/metal-organic framework composites with high stability in protein denaturing solvents were reported in this study.

Results

Encapsulation of enzyme in metal-organic frameworks (MOFs) via co-precipitation process was realized, and the generality of the synthesis was validated by using cytochrome c, horseradish peroxidase, and Candida antarctica lipase B as model enzymes. The stability of encapsulated enzyme was greatly increased after immobilization on MOFs. Remarkably, when exposed to protein denaturing solvents including dimethyl sulfoxide, dimethyl formamide, methanol, and ethanol, the enzyme/MOF composites still preserved almost 100% of activity. In contrast, free enzymes retained no more than 20% of their original activities at the same condition. This study shows the extraordinary protecting effect of MOF shell on increasing enzyme stability at extremely harsh conditions.

Conclusion

The enzyme immobilized in MOF exhibited enhanced thermal stability and high tolerance towards protein denaturing organic solvents.

Keywords

Biocatalysis / Biomineralization / Immobilization

References

[1] Bordusa F. Proteases in organic synthesis. Chem Rev, 2002, 102: 4817-4868.

[2] Bornscheuer U, Huisman G, Kazlauskas R, Lutz S, Moore J, Robins K. Engineering the third wave of biocatalysis. Nature, 2012, 485: 185-194.

[3] Brady D, Jordaan J. Advances in enzyme immobilisation. Biotechnol Lett, 2009, 31: 1639-1650.

[4] Cao S-L, Yue D-M, Li X-H, Smith TJ, Li N, Zong M-H, Wu H, Ma Y-Z, Lou W-Y. Novel nano-/micro-biocatalyst: soybean epoxide hydrolase immobilized on UiO-66-NH₂ MOF for efficient biosynthesis of enantiopure (R)-1, 2-octanediol in deep eutectic solvents. ACS Sustain Chem Eng, 2016, 4: 3586-3595.

[5] Chen K, Arnold FH. Enzyme engineering for nonaqueous solvents: random mutagenesis to enhance activity of subtilisin e in polar organic media. Nat Biotechnol, 1991, 9: 1073-1077.

[6] Chen Y, Lykourinou V, Vetromile C, Hoang T, Ming L-J, Larsen RW, Ma S. How can proteins enter the interior of a MOF? Investigation of cytochrome c translocation into a MOF consisting of mesoporous cages with microporous windows. J Am Chem Soc, 2012, 134: 13188-13191.

[7] Desai UR, Klibanov AM. Assessing the structural integrity of a lyophilized protein in organic solvents. J Am Chem Soc, 1995, 117: 3940-3945.

[8] Feng D, Liu T-F, Su J, Bosch M, Wei Z, Wan W, Yuan D, Chen Y-P, Wang X, Wang K, Lian X, Gu Z-Y, Park J, Zou X, Zhou H-C. Stable metal-organic frameworks containing single-molecule traps for enzyme encapsulation. Nat Commun, 2015, 6: 5979.

[9] Ferreira L, Gil MH, Dordick JS. Enzymatic synthesis of dextran-containing hydrogels. Biomaterials, 2002, 23: 3957-3967.

[10] Fujita K, MacFarlane DR, Forsyth M, Yoshizawa-Fujita M, Murata K, Nakamura N, Ohno H. Solubility and stability of cytochrome c in hydrated ionic liquids: effect of oxo acid residues and kosmotropicity. Biomacromolecules, 2007, 8: 2080-2086.

[11] Ge J, Lei J, Zare RN. Protein-inorganic hybrid nanoflowers. Nat Nanotechnol, 2012, 7: 428-432.

[12] Hanefeld U, Gardossi L, Magner E. Understanding enzyme immobilisation. Chem Soc Rev, 2009, 38: 453-468.

[13] He H, Han H, Shi H, Tian Y, Sun F, Song Y, Li Q, Zhu G. Construction of thermophilic lipase-embedded metal-organic frameworks via biomimetic mineralization: a biocatalyst for ester hydrolysis and kinetic resolution. ACS Appl Mater Interfaces, 2016, 8: 24517-24524.

[14] Ji X, Su Z, Wang P, Ma G, Zhang S. Integration of artificial photosynthesis system for enhanced electronic energy-transfer efficacy: a case study for solar-energy driven bioconversion of carbon dioxide to methanol. Small, 2016, 12: 4753-4762.

[15] Kim J, Grate JW, Wang P. Nanobiocatalysis and its potential applications. Trends Biotechnol, 2008, 26: 639-646.

[16] Kirk O, Borchert TV, Fuglsang CC. Industrial enzyme applications. Curr Opin Biotechnol, 2002, 13: 345-351.

[17] Lee C-H, Lang J, Yen C-W, Shih P-C, Lin T-S, Mou C-Y. Enhancing stability and oxidation activity of cytochrome c by immobilization in the nanochannels of mesoporous aluminosilicates. J Phys Chem B, 2005, 109: 12277-12286.

[18] Li P, Moon SY, Guelta MA, . Encapsulation of a nerve agent detoxifying enzyme by a mesoporous zirconium metal-organic framework engenders thermal and long-term stability. J Am Chem Soc, 2016, 138(26): 8052-8055.

[19] Li S, Dharmarwardana M, Welch RP, Ren Y, Thompson CM, Smaldone RA, Gassensmith JJ. Template-directed synthesis of porous and protective core–shell bionanoparticles. Angew Chem, 2016, 128: 10849-10854.

[20] Liang K, Carbonell C, Styles MJ, Ricco R, Cui J, Richardson JJ, Maspoch D, Caruso F, Falcaro P. Biomimetic replication of microscopic metal-organic framework patterns using printed protein patterns. Adv Mater, 2015, 27: 7293-7298.

[21] Liang K, Ricco R, Doherty CM, Styles MJ, Bell S, Kirby N, Mudie S, Haylock D, Hill AJ, Doonan CJ. Biomimetic mineralization of metal-organic frameworks as protective coatings for biomacromolecules. Nat Commun, 2015, 6: 7240.

[22] Liang H, Jiang S, Yuan Q, Li G, Wang F, Zhang Z, Liu J. Co-immobilization of multiple enzymes by metal coordinated nucleotide hydrogel nanofibers: improved stability and an enzyme cascade for glucose detection. Nanoscale, 2016, 8: 6071-6078.

[23] Liang K, Richardson JJ, Cui J, Caruso F, Doonan CJ, Falcaro P. Metal-organic framework coatings as cytoprotective exoskeletons for living cells. Adv Mater, 2016, 28: 7910-7914.

[24] Liang K, Coghlan CJ, Bell SG, Doonan C, Falcaro P. Enzyme encapsulation in zeolitic imidazolate frameworks: a comparison between controlled co-precipitation and biomimetic mineralisation. Chem Commun, 2016, 52: 473-476.

[25] Liu J, Yang Q, Li C. Towards efficient chemical synthesis via engineering enzyme catalysis in biomimetic nanoreactors. Chem Commun, 2015, 51: 13731-13739.

[26] Lykourinou V, Chen Y, Wang X-S, Meng L, Hoang T, Ming L-J, Musselman RL, Ma S. Immobilization of MP-11 into a mesoporous metal-organic framework, MP-11@ mesoMOF: a new platform for enzymatic catalysis. J Am Chem Soc, 2011, 133: 10382-10385.

[27] Lyu F, Zhang Y, Zare RN, Ge J, Liu Z. One-pot synthesis of protein-embedded metal-organic frameworks with enhanced biological activities. Nano Lett, 2014, 14: 5761-5765.

[28] Nilsson K, Mosbach K. Peptide synthesis in aqueous–organic solvent mixtures with α-chymotrypsin immobilized to tresyl chloride-activated agarose. Biotechnol Bioeng, 1984, 26: 1146-1154.

[29] Novak MJ, Pattammattel A, Koshmerl B, Puglia M, Williams C, Kumar CV. “Stable-on-the-table” enzymes: engineering the enzyme–graphene oxide interface for unprecedented kinetic stability of the biocatalyst. ACS Catal, 2015, 6: 339-347.

[30] Ono T, Goto M. Peroxidative catalytic behavior of cytochrome c solubilized in reverse micelles. Biochem Eng J, 2006, 28: 156-160.

[31] Pace C, Hermans J. The stability of globular protein. CRC Crit Rev Biochem, 1975, 3: 1-43.

[32] Serre C, Millange F, Thouvenot C, Nogues M, Marsolier G, Louer D, Ferey G. Very large breathing effect in the first nanoporous chromium(III)-based solids: MIL-53 or Cr^III(OH)·{O₂C− C₆H₄− CO₂}·{HO₂C− C₆H₄− CO₂H}_x·H₂O_y. J Am Soc Chem, 2002, 124: 13159-13526.

[33] Shi J, Wang X, Zhang S, Tang L, Jiang Z. Enzyme-conjugated ZIF-8 particles as efficient and stable pickering interfacial biocatalysts for biphasic biocatalysis. J Mater Chem B, 2016, 4: 2654-2661.

[34] Shieh F-K, Wang S-C, Yen C-I, Wu C-C, Dutta S, Chou L-Y, Morabito JV, Hu P, Hsu M-H, Wu KC-W. Imparting functionality to biocatalysts via embedding enzymes into nanoporous materials by a de novo approach: size-selective sheltering of catalase in metal-organic framework microcrystals. J Am Chem Soc, 2015, 137: 4276-4279.

[35] Wen L, Gao A, Cao Y, Svec F, Tan T, Lv Y. Layer-by-layer assembly of metal-organic frameworks in macroporous polymer monolith and their use for enzyme immobilization. Macromol Rapid Commun, 2016, 37: 551-557.

[36] Wu X, Yang C, Ge J, Liu Z. Polydopamine tethered enzyme/metal-organic framework composites with high stability and reusability. Nanoscale, 2015, 7: 18883-18886.

[37] Wu X, Ge J, Yang C, Hou M, Liu Z. Facile synthesis of multiple enzyme-containing metal-organic frameworks in a biomolecule-friendly environment. Chem Commun, 2015, 51: 13408-13411.

[38] Xu K, Griebenow K, Klibanov AM. Correlation between catalytic activity and secondary structure of subtilisin dissolved in organic solvents. Biotechnol Bioeng, 1997, 56: 485-491.

[39] Zhang Y, Wang F, Ju E, Liu Z, Chen Z, Ren J, Qu X. Metal-organic-framework-based vaccine platforms for enhanced systemic immune and memory response. Adv Funct Mater, 2016, 26: 6454-6461.

Funding

National Key Research and Development Plan of China(2016YFA0204300)