Free-Standing Metal Oxide Nanoparticle Superlattices Constructed with Engineered Protein Containers Show in Crystallo Catalytic Activity

Article abstract of DOI:10.1002/chem.201705061

The construction of defined nanostructured catalysts is challenging. In previous work, we established a strategy to assemble binary nanoparticle superlattices with oppositely charged protein containers as building blocks. Here, we show that these free-standing nanoparticle superlattices are catalytically active. The metal oxide nanoparticles inside the protein scaffold are accessible for a range of substrates and show oxidase-like and peroxidase-like activity. The stable superlattices can be reused for several reaction cycles. In contrast to bulk nanoparticle-based catalysts, which are prone to aggregation and difficult to characterize, nanoparticle superlattices based on engineered protein containers provide an innovative synthetic route to structurally defined heterogeneous catalysts with control over nanoparticle size and composition.

Full text of DOI:10.1002/chem.201705061

A Journal of
Accepted Article
Title: Free-standing metal oxide nanoparticle superlattices constructed
with engineered protein containers show in crystallo catalytic
activity
Authors: Tobias Beck, Marcel Lach, and Matthias Künzle
This manuscript has been accepted after peer review and appears as an
Accepted Article online prior to editing, proofing, and formal publication
of the final Version of Record (VoR). This work is currently citable by
using the Digital Object Identifier (DOI) given below. The VoR will be
published online in Early View as soon as possible and may be different
to this Accepted Article as a result of editing. Readers should obtain
the VoR from the journal website shown below when it is published
to ensure accuracy of information. The authors are responsible for the
content of this Accepted Article.
To be cited as: Chem. Eur. J. 10.1002/chem.201705061
Link to VoR: http://dx.doi.org/10.1002/chem.201705061
Supported by

10.1002/chem.201705061
Chemistry - A European Journal
COMMUNICATION
Free-standing metal oxide nanoparticle superlattices constructed
with engineered protein containers show in crystallo catalytic
activity
Marcel Lach, Matthias Künzle, and Tobias Beck*
Abstract: The construction of defined nanostructured catalysts is
challenging. In previous work, we established a strategy to assemble
binary nanoparticle superlattices with oppositely charged protein
containers as building blocks. Here, we show that these
free-standing nanoparticle superlattices are catalytically active. The
metal oxide nanoparticles inside the protein scaffold are accessible
for a range of substrates and show oxidase-like and peroxidase-like
activity. The stable superlattices can be reused for several reaction
cycles. In contrast to bulk nanoparticle-based catalysts, which are
prone to aggregation and difficult to characterize, nanoparticle
superlattices based on engineered protein containers provide an
innovative synthetic route to structurally defined heterogeneous
catalysts with control over nanoparticle size and compositions.
Protein scaffolds for nanoparticle superlattice assembly
offer an alternative route to catalytically active and well-defined
nanoparticle-based catalysts. Proteins can be assembled with
atomic precision into highly structured materials, either on
surfaces to form 2D films,[5] or as free-standing 3D crystals.[6]
Furthermore, the protein scaffold can be readily stabilized by
cross-linking of the building blocks in the protein crystals.[7]
Importantly, protein crystals show a high solvent content, thus a
high porosity, which enables ‘in crystallo’ enzymatic reactions,[8]
soaking of heavy-metal compounds for protein structure
determination,[9] or co-crystallization with insoluble dyes to
produce singlet oxygen.[10] In this work, we use protein
containers as building blocks to construct catalytically active
nanoparticle superlattices based on protein crystals. The pores
inherent to the protein containers and the channels inside the
protein crystal enable access to the nanoparticles, thus
overcoming previous challenges such as surface accessibility in
nanoparticle-based catalysts. Moreover, for the first time, large-
scale nanoparticle superlattices with dimensions up to several
hundred micrometers were prepared and utilized in catalysis.
We have recently established engineered ferritin
containers as building blocks for the construction of highly
ordered binary nanoparticle superlattices.[11] In the first step, the
protein containers are equipped with a number of charged amino
acids on the outer surface,[12] to effort two oppositely charged
protein containers.[11] Subsequently, metal oxide nanoparticle
synthesis inside these engineered containers, which have an
inner diameter of 7 nm and outer diameter of 12 nm, can be
readily carried out by exploiting the container pores (Fig. 1A).
For this work, we synthesized cerium oxide and iron oxide
nanoparticles inside the engineered protein containers. In the
final step, these nanoparticle-protein composites are used for
the construction of binary nanoparticle superlattices (Fig. 1B).
For the application of nanostructured materials in catalysis,
optics, electronics and sensor technology, a fine control over
material composition and morphology is required because of the
strong interplay between functionality and material structure.[1]
This holds particularly true for catalysis, where access to the
catalytically active surface is mandatory and
a defined
morphology and composition is required for reaction control.
However, it remains difficult to construct well-defined
nanomaterials, with precise distances between particles and
sufficient reaction space around them. Towards this end,
bottom-up approaches offer a solution. Here, nanoscale building
blocks such as nanoparticles are assembled into three-
dimensional materials with defined composition. Generally,
organic ligands or DNA linkers are used to construct
nanoparticle superlattices with diverse crystal symmetry, lattice
parameters and compositions.[2] However, certain drawbacks
exist. For nanoparticle assembly, the aforementioned
approaches require ligands that block the access to the
nanoparticles for catalysis, thus passivate the surface. This
could explain why there have been a number of reports on the
synthesis of nanoparticle superlattices,[2-3] but only a few studies
on their catalytic properties have been published, necessitating
the removal of the ligand scaffold by thermal decomposition.[4]
Moreover, nanoparticle superlattices based on DNA linkers
show only limited stability, which can be improved by elaborate
post-processing.[4c]
[*]
Marcel Lach, Matthias Künzle, Dr. Tobias Beck
RWTH Aachen University, Institute of Inorganic Chemistry;
JARA-SOFT (Researching Soft Matter); and I3TM
52074 Aachen (Germany)
E-mail: tobias.beck@ac.rwth-aachen.de
Figure 1. Strategy for the construction of binary metal oxide nanoparticles
superlattices. A) Nanoparticle synthesis inside the ferritin container cavity,
utilizing the container pores. B) Assembly of the nanoparticle-protein
composites to form highly ordered binary nanoparticle superlattices.
Supporting information for this article is given via a link at the end of
the document.
This article is protected by copyright. All rights reserved.

10.1002/chem.201705061
Chemistry - A European Journal
COMMUNICATION
For the current study, we mainly focused on cerium oxide
nanoparticles, which have over the years found application in
catalytic converters, solid oxide fuels cells, ultraviolet absorbers
and oxygen sensors.[13] Recently, cerium oxide nanoparticles,
also referred to as nanoceria, have been shown to have catalytic
properties similar to enzymes.[14] Due to its high biocompatibility,
nanoceria has been discussed as enzyme surrogate.[15] In
general, nanoparticles can be superior to enzymes in terms of
thermal stability and resistance against degradation. Moreover,
catalytic nanoparticles can introduce orthogonal reactivity, not
found in natural enzymatic reactions.[16] Therefore, the
construction of multifunctional nanoparticle catalysts could
produce materials with unparalleled catalytic activity and
diversity.
to investigate the oxidase-like activity, with oxygen as the
oxidant. For peroxidase-like activity, hydrogen peroxide was
added to the reaction mixture. Cerium oxide nanoparticles do
not show any strong absorption in the visible range, preventing
any spectral overlap with the dye molecule. The reaction was
followed by monitoring the absorbance maximum of the oxidized
TMB at 645 nm. Reaction without hydrogen peroxide shows a
turnover to the blue colored product within hours (Fig. 3A and B),
which is also observed in the corresponding UV-Vis spectra for
CeFtn(pos) and CeFtn(neg) (Fig. 3C). No background reaction was
observed without catalyst. Peroxidase-like activity in the
presence of hydrogen peroxide proceeds at a faster rate: the
blue color is visible within minutes (Fig. 3D).
As catalyst, cerium oxide nanoparticles were synthesized
inside the positively charged ferritin container Ftn(pos) and the
negatively charged container Ftn(neg) (see supporting information
for procedures) to yield CeFtn(pos) and CeFtn(neg). Powder X-ray
diffraction shows that the particles inside the containers are
CeO2, with the same crystal structure as commercially available
CeO2 nanoparticles (Fig. S1). The engineered protein containers
serve a threefold purpose: a) The ferritin containers are a
reaction template: The inner protein container cavity provides a
size-constraining vessel for the synthesis of nanoparticles, as
shown in several examples.[17] Moreover, protein containers with
encapsulated nanoparticles or enzymes have been applied for a
number of catalytic conversions.[18] b) The engineered ferritin
Figure 3. Catalysis in solution. Solution with 1 mM TMB and catalyst at 0 min
(a) and after one day (b). Absorption spectra of TMB after catalytic reaction
with CeO2 in Ftn(pos) and Ftn(neg): oxidase-like activity after 1 day (c) and
peroxidase-like activity with hydrogen peroxide present. Incubation time
15 min (d).
containers have
a charged outer surface, which enables
assembly of binary nanoparticles lattices[11] (Fig. 1), for the
construction of multifunctional materials. c) The containers’
inherent pores enable in situ synthesis of nanoparticles inside
the cavity. Importantly, after formation of the container-
nanoparticle composite, these pores provide access to the
nanoparticles for catalytic turnover. Although a number of
Kinetic parameters for the oxidase-like and peroxidase-like
activity of CeFtn(pos) and CeFtn(neg) in solution were determined
by varying the concentrations of TMB and hydrogen peroxide
(see supporting information for details, Figs. S2 and S3), and
are shown in Tab. 1.
protein
container-nanoparticle
composites
have
been
synthesized,[17] only very few catalytic systems have been
Table 1. Comparison of the kinetic parameters for oxidase-like (no
H2O2) or peroxidase-like activity (with H2O2) using the cerium
nanoparticles synthesized within the two protein containers, CeFtn.
kinetically characterized in solution.[17e,
Therefore, we
17f]
investigated the reactivity of the container-nanoparticle
composite both in solution and in the nanoparticle superlattice.
Building block
CeFtn(pos)
Substrate
TMB
vmax [µM s-1]
2.2 × 10-3
1.2 × 10-2
Km [mM]
1.54
CeFtn(pos)
TMB + H2O2
15.39
CeFtn(neg)
CeFtn(neg)
TMB
7.3 × 10-4
3.3 × 10-3
0.83
4.05
TMB + H2O2
For CeFtn(pos)
a higher activity is observed compared to
Figure 2. Catalytic oxidation of 3,3’,5,5’-tetramethylbenzidine (TMB) in
solution and in the nanoparticle superlattice using encapsulated cerium oxide
nanoparticles to form the oxidized substrate with blue color.
CeFtn(neg). This difference can be attributed to the fact that the
nanoparticle loading is higher for CeFtn(pos): After the sucrose
gradient centrifugation, the peak for the maximum absorption at
322 nm in the CeFtn(pos) sample is found at a higher fraction
number compared to CeFtn(neg) (Fig. S4). This shift indicates that
the CeO2 nanoparticles for Ftn(pos) have a higher mass and thus
3,3’,5,5’-tetramethylbenzidine (TMB),
a substrate for horse-
radish peroxidase commonly used in bioassays,[19] was used to
investigate the catalytic oxidation to the readily traceable blue
oxidation product (Fig. 2). The reaction was started by mixing
catalyst, either CeFtn(pos) or CeFtn(neg), with the TMB substrate.
Reactions with TMB under aerobic atmosphere were conducted
Ftn(pos) has a higher loading efficiency compared with Ftn(neg)
The higher loading efficiency in the final samples (same protein
concentration) can also be observed with UV-Vis absorption
spectroscopy (Fig. S5).[11] Previous syntheses of the cerium
.
This article is protected by copyright. All rights reserved.

10.1002/chem.201705061
Chemistry - A European Journal
COMMUNICATION
oxide nanoparticles also showed using transmission electron
microscopy that CeFtn(pos) yielded slightly larger nano-
particles.[11] Notably, the reaction velocity in the presence of
hydrogen peroxide (peroxidase-like activity) is faster for
CeFtn(pos) and CeFtn(neg) each, possibly due to different reaction
pathways. For peroxidase-like activity, the cerium oxide
20]
nanoparticles likely catalyze a Fenton-like reaction,[14,
with
conversion of hydrogen peroxide to hydroxyl and superoxide
radicals that readily diffuse out of the container and react with
the substrate. For the oxidase-like activity (no hydrogen
peroxide present), the substrate needs to diffuse into the
container to be oxidized on the particle’s surface. A similar trend
in reaction velocity and Km with and without hydrogen peroxide
was observed for ferritin loaded with platinum nanoparticles.[17f]
Comparing the kinetic parameters for CeFtn composites with
cerium oxide nanoparticles coated with polymer ligands[21]
shows that the oxidase-like reaction velocity for the protein-
nanoparticle composites is considerable slower (factor 103),
because the substrates need to diffuse through the container
pores into the cavity for turnover, and out after the reaction.
Interestingly, the Michaelis constants are rather similar
compared to polymer-coated nanoparticles, indicating a similar
binding affinity to the nanoparticle surface for both types of
catalysts.[21] We also characterized the activity of the negatively
charged container loaded with iron oxide nanoparticles,
FeFtn(neg), towards catalytic oxidation of TMB. As expected,
these protein-nanoparticle composites do not show any oxidase-
like activity. In contrast to CeFtn, additional hydrogen peroxide is
required for the oxidation of TMB with FeFtn(neg) (Figs. S6 and
S7). Kinetic parameters are shown in Tab. S1 and show a
Figure 4. a) CeFtn(pos) / CeFtn(neg) crystals b) CeFtn(pos) / FeFtn(neg) crystal
c) CeFtn(pos) / CeFtn(neg) crystal after 20 minutes incubation with TMB substrate
d) CeFtn(pos) / FeFtn(neg) crystal after 20 minutes incubation with TMB substrate.
Scale bar is 100 µm. e) UV-Vis absorption spectrum of a CeFtn(pos) / CeFtn(neg)
crystal cut into thin slices.
Further investigations on in crystallo activity were carried out
with nanoparticle superlattices constructed with iron oxide
nanoparticles, FeFtn(neg). Incubation with TMB under aerobic
atmosphere did not show any reaction, thus no oxidase-like
activity (Fig. S11A), which is in accordance with the experiments
with the protein-nanoparticle composites in solution (see above).
Incubation of the iron oxide nanoparticle superlattice with TMB in
presence of H2O2 showed oxidation to the colored product, thus
peroxidase activity (Fig. S11B), as observed in solution.
As
a
step towards multifunctional materials, we
constructed nanoparticle superlattices with cerium and iron
oxide nanoparticles. These binary superlattices showed as
expected oxidase-like activity due to the cerium oxide
nanoparticles (Fig. 4B and D). The iron oxide nanoparticles do
not hamper the activity of the cerium oxide nanoparticles. Here,
a synergistic effect is not present, because only CeFtn shows
oxidase activity whereas FeFtn does not (see also Fig. 3). The
combination of two different nanoparticles in the protein scaffold
does not influence the activity of the single nanoparticle type. A
more detailed analysis of the kinetic parameters requires a
larger amount of materials, which will be produced using batch
crystallization techniques. Importantly, by measuring the product
concentration in the supernatant, we could show that the
crystalline samples are stable and enable several cycles of
catalytic turnover with high activity remaining in each cycle (Fig.
S12). Moreover, this is an advancement compared to previous
reports on catalytically active nanoparticle superlattices.[4c] In the
protein scaffold, the nanoparticles are not passivated nor is the
particle superlattice inactivated by the substrates.
To investigate the substrate scope of the oxidation reaction,
several dyes with different size were tested for conversion with
CeFtn(pos) / CeFtn(neg) crystals as catalyst without hydrogen
peroxide present (see supporting information for details). Here,
FeFtn crystals were not used due to the strong color of iron
oxide nanoparticle crystals. The dye molecules are readily
converted inside the nanoparticle superlattice as evidenced by
deep coloration of the crystals incubated with substrate solution
(Fig. S13-S17). Interestingly, the substrates are slightly larger in
thickness than the container pore size at the three-fold channel
(TMB 5 Å vs. pore 4 Å, Fig. S18). Obviously, the flexibility of the
ferritin pores, observed already for incorporation of large
molecules such as organometallic complexes into the ferritin
cavity,[22] enables the substrates accessing the nanoparticles. By
similar reaction rate compared with CeFtn(pos) and CeFtn(neg)
.
We were particularly interested if the catalytic activity
observed in solution is preserved within the nanoparticle
superlattices. With the oppositely charged protein containers as
building blocks, nanoparticles were assembled (Fig. 1B) to yield
crystalline samples with dimensions up to several hundred
micrometers (Fig. S8), using crystallization conditions previously
established.[11] Before the materials were subjected to catalytic
investigations, the protein scaffold was stabilized by incubation
with a dialdehyde that cross-links the side chains inside the
crystals. The nanoparticle crystals prepared in this way can be
readily manipulated. Moreover, they are stable in reaction
solutions for testing catalytic activity. As first samples, we used
nanoparticle superlattices with cerium oxide nanoparticles.
Either Ftn(pos) or Ftn(neg), or both containers are filled with
nanoparticles and assembled into highly ordered superlattices.
Importantly, incubation of the crystalline materials with a solution
containing TMB shows that the cerium oxide nanoparticles are
catalytically active within the protein matrix and show a deep
coloration within minutes for the CeFtn(pos) / CeFtn(neg) crystals
(Fig. 4A, C, E). This can also be observed for crystals with either
container filled with CeO2 (Fig. S9). Peroxidase activity is also
observed for all three nanoparticle superlattices (Fig. S10). The
crystal concentrates the catalytically active nanoparticles in a
confined space. Therefore, the turnover is more quickly visible in
the nanoparticle superlattice compared to solution. After the
reaction, the converted substrate can diffuse out of the crystal.
This article is protected by copyright. All rights reserved.

10.1002/chem.201705061
Chemistry - A European Journal
COMMUNICATION
Qin, S. Li, X. Chen, Nanoscale 2012, 4, 66-75; d) V. F. Puntes, P. Gorostiza, D.
M. Aruguete, N. G. Bastus, a. P. Alivisatos, Nat. Mater. 2004, 3, 263-268; e) A.
K. Gupta, M. Gupta, Biomaterials 2005, 26, 3995-4021.
a) E. V. Shevchenko, D. V. Talapin, N. A. Kotov, S. O'Brien, C. B. Murray,
Nature 2006, 439, 55-59; b) X. Ye, J. Chen, C. B. Murray, J. Am. Chem. Soc.
2011, 133, 2613-2620; c) R. J. Macfarlane, M. R. Jones, B. Lee, E. Auyeung,
C. A. Mirkin, Science 2013, 341, 1222-1225; d) C. Zhang, R. J. Macfarlane, K.
L. Young, C. H. J. Choi, L. Hao, E. Auyeung, G. Liu, X. Zhou, C. A. Mirkin,
Nat. Mater. 2013, 12, 741-746.
a) S. Y. Park, A. K. R. Lytton-Jean, B. Lee, S. Weigand, G. C. Schatz, C.
Mirkin, Nature 2008, 451, 553-556; b) D. Nykypanchuk, M. M. Maye, D. van
der Lelie, O. Gang, Nature 2008, 451, 549-552; c) M. A. Kostiainen, P.
Hiekkataipale, A. Laiho, V. Lemieux, J. Seitsonen, J. Ruokolainen, P. Ceci,
Nat. Nanotechnol. 2013, 8, 52-56.
a) Y. Kang, X. Ye, J. Chen, L. Qi, R. E. Diaz, V. Doan-Nguyen, G. Xing, C. R.
Kagan, J. Li, R. J. Gorte, et al., J. Am. Chem. Soc. 2013, 135, 1499-1505; b) J.
Li, Y. Wang, T. Zhou, H. Zhang, X. Sun, J. Tang, L. Zhang, A. M. Al-Enizi, Z.
Yang, G. Zheng, J. Am. Chem. Soc. 2015, 137, 14305-14312; c) E. Auyeung,
W. Morris, J. E. Mondloch, J. T. Hupp, O. K. Farha, C. A. Mirkin, J. Am.
Chem. Soc. 2015, 137, 1658-1662.
comparing the reaction rate of the substrates to the reaction rate
of TMB in the crystals, it can be concluded that
3,3’-diaminobenzidin and 3-amino-9-ethylcarbazole (Fig. S16
and S17) show a similar reaction rate as TMB, as coloration of
the crystals is observed within minutes. O-phenyldiamine,
5-aminosalicylic acid and o-dianisidine react considerably slower
(Fig. S13 – S15), because coloration is only clearly visible after
one day. Because there is no correlation of substrate size and
reactivity, the reactivity does apparently not depend on the size
of the substrates used in this study. Nevertheless, the container
pores and the crystal channels (diameter of about 10 Å, Fig.
S19) could offer a potential filter effect, e.g. with regard to the
size or polarity of the substrates, further exploited by
engineering the containers’ pores and crystal lattice channels.
Our results demonstrate that engineered protein
containers can function as building blocks for well-defined
heterogeneous catalysts based on metal oxide nanoparticles.
The nanoparticles show oxidase-like and peroxidase-like activity
inside the crystalline material. The nanoparticle materials are
stable in solution and can be produced with lateral dimensions
up to several hundred micrometers. The protein scaffold
provides stability to the nanoparticles but ensures access to the
nanoparticles via the channels within the crystal and the protein
[2]
[3]
[4]
[5]
[6]
a) P. Ringler, Science 2003, 302, 106-109; b) S. Gonen, F. DiMaio, T. Gonen,
D. Baker, Science 2015, 348, 1365-1368.
a) P. A. Sontz, J. B. Bailey, S. Ahn, F. A. Tezcan, J. Am. Chem. Soc. 2015, 137,
11598-11601; b) C. J. Lanci, C. M. Macdermaid, S.-g. Kang, R. Acharya, B.
North, X. Yang, Proc. Natl. Acad. Sci. U.S.A. 2012, 109, 7304-7309.
J. J. Roy, T. E. Abraham, Chem. Rev. 2004, 104, 3705-3721.
a) A. Mozzarelli, G. L. Rossi, Annu. Rev. Biophys. Biomol. Struct. 1996, 25,
343-365; b) A. L. Margolin, M. A. Navia, Angew. Chem. Int. Ed. 2001, 40,
2204-2222.
a) T. Beck, A. Krasauskas, T. Gruene, G. M. Sheldrick, Acta Crystallogr. Sect.
D 2008, 64, 1179-1182; b) A. C. W. Pike, E. F. Garman, T. Krojer, F. von
Delft, E. P. Carpenter, Acta Crystallogr. Sect. D 2016, 72, 303-318.
[7]
[8]
[9]
[10] J. Mikkilä, E. Anaya-Plaza, V. Liljeström, J. R. Caston, T. Torres, A. de la
Escosura, M. A. Kostiainen, ACS Nano 2016, 10, 1565-1571.
container pores. In this way,
a long-standing challenge,
nanoparticle surface accessibility in nanoparticle superlattices, is
overcome. In addition to providing stability to the nanoparticle
lattice, the protein scaffold can also impart biocompatibility of the
material, important for applications in the biomedical field. Here,
the utilization of nanoparticles is still hampered by their
compatibility and stability in biological systems. The presented
binary system enables a fine control over the composition of the
heterogeneous catalyst. Along these lines, binary superlattices
built from nanoparticles, also other than the ones used in the
current study, with different or complementary catalytic
functionality could catalyze cascade reactions within the protein
scaffold.
[11] M. Künzle, T. Eckert, T. Beck, J. Am. Chem. Soc. 2016, 138, 12731-12734.
[12] T. Beck, S. Tetter, M. Künzle, D. Hilvert, Angew. Chem. Int. Ed. 2015, 54,
937-940.
[13] a) M. Lei, T. Z. Yang, W. J. Wang, K. Huang, R. Zhang, X. L. Fu, H. J. Yang,
Y. G. Wang, W. H. Tang, Int. J. Hydrogen Energy 2013, 38, 205-211; b) M.
Aguirre, M. Paulis, J. R. Leiza, Journal of Materials Chemistry A 2013, 1,
3155-3155; c) I. Shitanda, S. Mori, M. Itagaki, Anal. Sci. 2011, 27, 1049-1052;
d) J. Kašpar, P. Fornasiero, N. Hickey, Catal. Today 2003, 77, 419-449.
[14] C. Xu, X. Qu, NPG Asia Mater. 2014, 6, e90.
[15] X. Liu, W. Wei, Q. Yuan, X. Zhang, N. Li, Y. Du, G. Ma, C. Yan, D. Ma,
Chem. Commun. 2012, 48, 3155-3157.
[16] L. L. Chng, N. Erathodiyil, J. Y. Ying, Acc. Chem. Res. 2013, 46, 1825-1837.
[17] a) F. C. Meldrum, B. R. Heywood, S. Mann, Science 1992, 257, 522-523; b) R.
M. Kramer, C. Li, D. C. Carter, M. O. Stone, R. R. Naik, J. Am. Chem. Soc.
2004, 126, 13282-13286; c) M. Uchida, M. L. Flenniken, M. Allen, D. A.
Willits, B. E. Crowley, S. Brumfield, A. F. Willis, L. Jackiw, M. Jutila, M. J.
Young, T. Douglas, J. Am. Chem. Soc. 2006, 128, 16626-16633; d) C. A. Butts,
J. Swift, S.-G. Kang, L. Di Costanzo, D. W. Christianson, J. G. Saven, I. J.
Dmochowski, Biochemistry 2008, 47, 12729-12739; e) Z. Varpness, J. W.
Peters, M. Young, T. Douglas, Nano Lett. 2005, 5, 2306-2309; f) J. Fan, J.-J.
Yin, B. Ning, X. Wu, Y. Hu, M. Ferrari, G. J. Anderson, J. Wei, Y. Zhao, G.
Nie, Biomaterials 2011, 32, 1611-1618.
[18] a) A. Liu, C. H. H. Traulsen, J. J. L. M. Cornelissen, ACS Catalysis 2016, 6,
3084-3091; b) A. Liu, L. Yang, C. H. H. Traulsen, J. J. L. M. Cornelissen,
Chem. Commun. 2017, 53, 7632-7634; c) P. C. Jordan, D. P. Patterson, K. N.
Saboda, E. J. Edwards, H. M. Miettinen, G. Basu, M. C. Thielges, T. Douglas,
Nature Chemistry 2015, 8, 1-7; d) Y. Azuma, R. Zschoche, M. Tinzl, D.
Hilvert, Angew. Chem. Int. Ed. 2016, 55, 1531-1534; e) S. Tetter, D. Hilvert,
Angew. Chem. Int. Ed. 2017, 10.1002/anie.201708530, 1-5.
Acknowledgements
We thank Prof. Ulrich Schwaneberg for generous support with
regard to protein production and Prof. Ulrich Simon for general
support and helpful discussions. This work was generously
supported by
Chemischen Industrie),
a
Liebig scholarship to T.B. (Fonds der
doctoral scholarship to M.L.
a
(Cusanuswerk), a doctoral scholarship to M.K. (Fonds der
Chemischen Industrie), and the Excellence Initiative of the
German federal and state governments (I3TM Seed Fund grant
and I3TM Step2Project grant to T.B.).
[19] A. Frey, B. Meckelein, D. Externest, M. A. Schmidt, J. Immunol. Methods
2000, 233, 47-56.
[20] E. G. Heckert, S. Seal, W. T. Self, Environ. Sci. Technol. 2008, 42, 5014-5019.
[21] A. Asati, S. Santra, C. Kaittanis, S. Nath, J. M. Perez, Angew. Chem. Int. Ed.
2009, 48, 2308-2312.
[22] a) S. Abe, K. Hirata, T. Ueno, K. Morino, N. Shimizu, M. Yamamoto, M.
Takata, E. Yashima, Y. Watanabe, J. Am. Chem. Soc. 2009, 131, 6958-6960;
b) S. Zhang, J. Zang, H. Chen, M. Li, C. Xu, G. Zhao, Small 2017, 13,
1701045-1701045.
Keywords: protein engineering • protein container •
nanocatalysis • nanoparticles • nanoparticle superlattice
[1]
a) D. V. Talapin, J.-S. Lee, M. V. Kovalenko, E. V. Shevchenko, Chem. Rev.
2010, 110, 389-458; b) A.-H. Lu, W. Schmidt, N. Matoussevitch, H.
Bönnemann, B. Spliethoff, B. Tesche, E. Bill, W. Kiefer, F. Schüth, Angew.
Chem. Int. Ed. 2004, 43, 4303-4306; c) L. Jiang, Y. Sun, F. Huo, H. Zhang, L.
This article is protected by copyright. All rights reserved.

10.1002/chem.201705061
Chemistry - A European Journal
COMMUNICATION
COMMUNICATION
Marcel, Lach, Matthias Künzle, and
Tobias Beck *
Page No. – Page No.
Free-standing metal oxide
nanoparticle superlattices
constructed with engineered protein
containers show in crystallo catalytic
activity
Nanoparticle superlattices for catalysis: Engineered protein containers can
assemble metal oxide nanoparticles into crystalline three-dimensional materials.
The nanoparticles, which are accessible within the protein scaffold through the
protein container pores and crystal channels, catalyze the conversion of a range of
dye substrates.
This article is protected by copyright. All rights reserved.