ACS Publications. Most Trusted. Most Cited. Most Read
My Activity

Figure 1Loading Img

Immobilization of Enzymes in Mesoporous Silica Particles: Protein Concentration and Rotational Mobility in the Pores

View Author Information
Department of Chemistry and Chemical Engineering, Chalmers University of Technology, Kemivägen 10, SE-41296 Gothenburg, Sweden
*E-mail: [email protected]. Tel: +46317723052.
Cite this: J. Phys. Chem. B 2017, 121, 12, 2575–2583
Publication Date (Web):March 3, 2017
Copyright © 2017 American Chemical Society

    Article Views





    Other access options
    Supporting Info (1)»


    Abstract Image

    Enzyme immobilization in porous silica particles is used to improve enzyme function in biocatalytic applications. Here, we study the effective protein concentration and rotational mobility of lipase and bovine serum albumin in the pores, when confined in five types of mesoporous silica particles with different pore and particle sizes, exploiting the intrinsic UV–vis absorption and fluorescence anisotropy of the tryptophan residues. For all investigated combinations of proteins and particles, the steady-state anisotropy is higher than for the same protein in free solution, indicating a slower protein rotation inside the pores. The retardation is stronger in more narrow pores, but the proteins can still move, and there is no dependence on the particle size. The average number of proteins per particle, Nprot, varies with the particle diameter, D, as NprotD2.95±0.02 for both proteins, which is close to the scaling D3.0±0.1 for the available pore volume. This observation indicates that both proteins are distributed evenly throughout the particles and rules out that the proteins are only externally bound to the particle surface. Secondly, the concentration of the protein in the pores depends on the pore and protein size but not on the particle size and corresponds to volume fractions in the range of 20–60%.

    Read this article

    To access this article, please review the available access options below.

    Get instant access

    Purchase Access

    Read this article for 48 hours. Check out below using your ACS ID or as a guest.


    Access through Your Institution

    You may have access to this article through your institution.

    Your institution does not have access to this content. You can change your affiliated institution below.

    Supporting Information

    Jump To

    The Supporting Information is available free of charge on the ACS Publications website at DOI: 10.1021/acs.jpcb.7b00562.

    • Calculation of pore filling and number of proteins per particle, polarized fluorescence emission spectra in the absence and presence of particles, calculation of average pore volume per particle versus particle size, rotation of the whole MPS particle (θpart) and its contribution in anisotropy (PDF)

    Terms & Conditions

    Most electronic Supporting Information files are available without a subscription to ACS Web Editions. Such files may be downloaded by article for research use (if there is a public use license linked to the relevant article, that license may permit other uses). Permission may be obtained from ACS for other uses through requests via the RightsLink permission system:

    Cited By

    This article is cited by 33 publications.

    1. Mary Lenertz, Qiaobin Li, Zoe Armstrong, Allison Scheiwiller, Gigi Ni, Jien Wang, Li Feng, Austin MacRae, Zhongyu Yang. Magnetic Multienzyme@Metal–Organic Material for Sustainable Biodegradation of Insoluble Biomass. ACS Applied Materials & Interfaces 2024, 16 (9) , 11617-11626.
    2. Biswajit Hansda, Shailja Mishra, Ankit Ghosh, Basudev Das, Tirtha Biswas, Tanay K. Mondal, Bhavya Srivastava, Sneha Mondal, Dipika Roy, Bhabatosh Mandal. Chemically Bonded Pepsin via Its Inert Center to Diazo Functionalized Silica Gel through Multipoint Attachment Mode: A Way of Restoring Biocatalytic Sustainability over “Wider pH” Range. Langmuir 2024, 40 (4) , 2146-2164.
    3. Fanrui Sha, Haomiao Xie, Florencia A. Son, Kevin S. Kim, Wei Gong, Shengyi Su, Kaikai Ma, Xiaoliang Wang, Xingjie Wang, Omar K. Farha. Rationally Tailored Mesoporous Hosts for Optimal Protein Encapsulation. Journal of the American Chemical Society 2023, 145 (30) , 16383-16390.
    4. Nay San Lin, Kota Hirayama, Masaki Kitamura, Shinji Koide, Hiromasa Kitajima, Takunori Harada, Shigeki Mayama, Kazuo Umemura. Fabrication of a Floatable Micron-Sized Enzyme Device Using Diatom Frustules. ACS Omega 2023, 8 (23) , 21145-21153.
    5. Shailja Mishra, Biswajit Hansda, Ankit Ghosh, Sneha Mondal, Bhabatosh Mandal, Pallavi Kumari, Basudev Das, Tanay Kumar Mondal, Tirtha Biswas. Multipoint Immobilization at Inert Center of Papain on Homo-Functional Diazo-Activated Silica Support: A Way of Restoring “Above Room-Temperature” Bio-Catalytic Sustainability. Langmuir 2023, 39 (16) , 5710-5726.
    6. Yongxin Bu, Lingyan Hu, Wei Feng. d-Amino Acid Oxidase Immobilized on Pt Nanoparticle-Loaded Porous SiO2 Nanospheres Coated with a Zirconium-Based Coordination Polymer for Catalytic Deamination of d-Alanine. ACS Applied Nano Materials 2021, 4 (11) , 12373-12381.
    7. Donya Valikhani, Juan M. Bolivar, Joelle N. Pelletier. An Overview of Cytochrome P450 Immobilization Strategies for Drug Metabolism Studies, Biosensing, and Biocatalytic Applications: Challenges and Opportunities. ACS Catalysis 2021, 11 (15) , 9418-9434.
    8. Milene Zezzi do Valle Gomes, Pegah S. Nabavi Zadeh, Anders E. C. Palmqvist, Björn Åkerman. Spatial Distribution of Enzymes Immobilized in Mesoporous Silicas for Biocatalysis. ACS Applied Nano Materials 2019, 2 (11) , 7245-7254.
    9. Chengmin Hou, Nicolas Ghéczy, Daniel Messmer, Katarzyna Szymańska, Jozef Adamcik, Raffaele Mezzenga, Andrzej B. Jarzębski, Peter Walde. Stable Immobilization of Enzymes in a Macro- and Mesoporous Silica Monolith. ACS Omega 2019, 4 (4) , 7795-7806.
    10. Claudia Marschelke, Martin Müller, Dorina Köpke, Anke Matura, Marco Sallat, Alla Synytska. Hairy Particles with Immobilized Enzymes: Impact of Particle Topology on the Catalytic Activity. ACS Applied Materials & Interfaces 2019, 11 (1) , 1645-1654.
    11. Seong-Min Jo, Frederik R. Wurm, Katharina Landfester. Biomimetic Cascade Network between Interactive Multicompartments Organized by Enzyme-Loaded Silica Nanoreactors. ACS Applied Materials & Interfaces 2018, 10 (40) , 34230-34237.
    12. Pegah S. Nabavi Zadeh, Milene Zezzi do Valle Gomes, Björn Åkerman, Anders E. C. Palmqvist. Förster Resonance Energy Transfer Study of the Improved Biocatalytic Conversion of CO2 to Formaldehyde by Coimmobilization of Enzymes in Siliceous Mesostructured Cellular Foams. ACS Catalysis 2018, 8 (8) , 7251-7260.
    13. Alessandra Scano, Edmond Magner, Martina Pilloni, Luciano Atzori, Marzia Fantauzzi, Sawssen Slimani, Davide Peddis, Gonzalo Garcia Fuentes, Guido Ennas. Combining high energy ball milling and liquid crystal templating method to prepare magnetic ordered mesoporous silica. A physico-chemical investigation. Physical Chemistry Chemical Physics 2024, 26 (17) , 13020-13033.
    14. Alessandra Machado Baron, Ricardo de Sousa Rodrigues, Luis Guilherme Giannina Sante, Jocácia Muriele de Miranda Kister, Valéria Marta Gomes do Nascimento, Alesandro Bail. Metal-organic framework based on iron and terephthalic acid as a multiporous support for lipase Burkholderia lata LBBIO-BL02 and its potential for biocatalysis. Biocatalysis and Biotransformation 2023, 41 (5) , 332-343.
    15. Rocio Villa, Susana Nieto, Antonio Donaire, Pedro Lozano. Direct Biocatalytic Processes for CO2 Capture as a Green Tool to Produce Value-Added Chemicals. Molecules 2023, 28 (14) , 5520.
    16. Karen Rodríguez-Núñez, Claudia Bernal, Juan M. Bolívar. The enzyme, the support, and the immobilization strategy: The key findings to a desirable biocatalyst. 2023, 1-16.
    17. Sahin Demirci, Nurettin Sahiner. Thermo‐responsive macroporous p( NIPAM ) cryogel affords enhanced thermal stability and activity for ɑ‐glucosidase enzyme by entrapping in situ. The Canadian Journal of Chemical Engineering 2022, 100 (12) , 3575-3587.
    18. Aleksandrina M. Sulman, Angela K. Haskell, Boris B. Tikhonov, Olga V. Grebennikova, Aleksandr I. Sidorov, Lyudmila M. Bronstein, Valentina G. Matveeva. Larger pores dramatically enhance activity of an immobilized enzyme in mesoporous magnetic silica. Microporous and Mesoporous Materials 2022, 341 , 112092.
    19. Saadat Majeed, Rahat Nawaz, Tahir Rasheed, Muhammad Bilal. Silica-based nanomaterials in biocatalysis. 2022, 171-188.
    20. Ivalina Trendafilova, Hristina Lazarova, Ralitsa Chimshirova, Boryana Trusheva, Neli Koseva, Margarita Popova. Novel kaempferol delivery systems based on Mg-containing MCM-41 mesoporous silicas. Journal of Solid State Chemistry 2021, 301 , 122323.
    21. Maria Fecková, Judit Tóth, Petr Šálek, Alena Španová, Daniel Horák, Quazi T. H. Shubhra, Aleš Kovařík, János Gyenis, Bohuslav Rittich. Capture of DNAs by magnetic hypercrosslinked poly(styrene-co-divinylbenzene) microspheres. Journal of Materials Science 2021, 56 (9) , 5817-5829.
    22. Carminna Ottone, Oscar Romero, Paulina Urrutia, Claudia Bernal, Andrés Illanes, Lorena Wilson. Enzyme Biocatalysis and Sustainability. 2021, 383-413.
    23. Shohreh Ariaeenejad, Farzaneh Jokar, Parvin Hadian, Leila Ma'mani, Sajjad Gharaghani, Masood Fereidoonnezhad, Ghasem Hosseini Salekdeh. An efficient nano-biocatalyst for lignocellulosic biomass hydrolysis: Xylanase immobilization on organically modified biogenic mesoporous silica nanoparticles. International Journal of Biological Macromolecules 2020, 164 , 3462-3473.
    24. Ivalina Trendafilova, Judith Mihály, Denitsa Momekova, Ralitsa Chimshirova, Hristina Lazarova, Georgi Momekov, Margarita Popova. Antioxidant activity and modified release profiles of morin and hesperetin flavonoids loaded in Mg- or Ag-modified SBA-16 carriers. Materials Today Communications 2020, 24 , 101198.
    25. Agnieszka Chrzanowska, Anna Derylo-Marczewska, Malgorzata Wasilewska. Mesocellular Silica Foams (MCFs) with Tunable Pore Size as a Support for Lysozyme Immobilization: Adsorption Equilibrium and Kinetics, Biocomposite Properties. International Journal of Molecular Sciences 2020, 21 (15) , 5479.
    26. Nur Fathiah Mokhtar, Raja Noor Zaliha Raja Abd. Rahman, Noor Dina Muhd Noor, Fairolniza Mohd Shariff, Mohd Shukuri Mohamad Ali. The Immobilization of Lipases on Porous Support by Adsorption and Hydrophobic Interaction Method. Catalysts 2020, 10 (7) , 744.
    27. Sukriti Gakhar, Subhash H. Risbud, Marjorie L. Longo. Structure retention of silica gel-encapsulated bacteriorhodopsin in purple membrane and in lipid nanodiscs. Colloids and Surfaces B: Biointerfaces 2020, 186 , 110680.
    28. Ameneh Ahmadi, Tahereh Sedaghat, Roya Azadi, Hossein Motamedi. Magnetic Mesoporous Silica Nanocomposite Functionalized with Palladium Schiff Base Complex: Synthesis, Characterization, Catalytic Efficacy in the Suzuki–Miyaura Reaction and α-Amylase Immobilization. Catalysis Letters 2020, 150 (1) , 112-126.
    29. Yan Li, Jia-Qin Wu, Neng-Bing Long, Rui-Feng Zhang. Efficient immobilization of phospholipase D on novel polymer supports with hierarchical pore structures. International Journal of Biological Macromolecules 2019, 141 , 60-67.
    30. Hui Li, Yanxiong Pan, Jasmin Farmakes, Feng Xiao, Guodong Liu, Bingcan Chen, Xiao Zhu, Jiajia Rao, Zhongyu Yang. A sulfonated mesoporous silica nanoparticle for enzyme protection against denaturants and controlled release under reducing conditions. Journal of Colloid and Interface Science 2019, 556 , 292-300.
    31. Zahra Fathi, Esmail Doustkhah, Golamhossein Ebrahimipour, Farshad Darvishi. Noncovalent Immobilization of Yarrowia lipolytica Lipase on Dendritic-Like Amino Acid-Functionalized Silica Nanoparticles. Biomolecules 2019, 9 (9) , 502.
    32. Diganta Saikia, Juti Rani Deka, Cheng-En Wu, Yung-Chin Yang, Hsien-Ming Kao. pH responsive selective protein adsorption by carboxylic acid functionalized large pore mesoporous silica nanoparticles SBA-1. Materials Science and Engineering: C 2019, 94 , 344-356.
    33. Pegah S. Nabavi Zadeh, Milene Zezzi do Valle Gomes, Maria Abrahamsson, Anders E. C. Palmqvist, Björn Åkerman. Measuring viscosity inside mesoporous silica using protein-bound molecular rotor probe. Physical Chemistry Chemical Physics 2018, 20 (36) , 23202-23213.

    Pair your accounts.

    Export articles to Mendeley

    Get article recommendations from ACS based on references in your Mendeley library.

    Pair your accounts.

    Export articles to Mendeley

    Get article recommendations from ACS based on references in your Mendeley library.

    You’ve supercharged your research process with ACS and Mendeley!

    STEP 1:
    Click to create an ACS ID

    Please note: If you switch to a different device, you may be asked to login again with only your ACS ID.

    Please note: If you switch to a different device, you may be asked to login again with only your ACS ID.

    Please note: If you switch to a different device, you may be asked to login again with only your ACS ID.

    Your Mendeley pairing has expired. Please reconnect