Other — Shielding against Unfolding by Embedding Enzymes in Metal-Organic Frameworks via a de Novo Approach

9 measurement groups · 38 results

Reported values remain attached to the sample, method, conditions, extraction quality and source location that produced them.

Catalase H2O2 decomposition by FOX assay

CAT@MCF · Powder

CAT@MCF incubated under analogous FOX assay conditions with urea or heat treatment.

Context: Non-MOF loose-confinement comparison
Measurement source: S10 · Examinations of catalytic activities · Table S1

Property	Reported value	Normalised value	Uncertainty	Origin and quality	Source
kobs after 0.05 M urea	(1.10 +/- 0.14) x 10^-3 s^-1	—	+/- 0.00014 s-1	SI Table Exact Reported	S16 · Table Section · Table S1
kobs after 6 M urea	(1.00 +/- 0.90) x 10^-6 s^-1	—	+/- 0.0000009 s-1	SI Table Exact Reported	S16 · Table Section · Table S1
kobs after 80 deg C treatment	(4.50 +/- 1.40) x 10^-5 s^-1	—	+/- 0.000014 s-1	SI Table Exact Reported	S16 · Table Section · Table S1
kobs after 8 M urea	(3.70 +/- 0.20) x 10^-6 s^-1	—	+/- 0.0000002 s-1	SI Table Exact Reported	S16 · Table Section · Table S1
kobs, buffer without urea	(1.53 +/- 0.04) x 10^-2 s^-1	—	+/- 0.0004 s-1	SI Table Exact Reported	S16 · Table Section · Table S1
Catalase loading in CAT@MCF	~8.5 wt %	—	—	Text Approximate	S8 · Synthesis of CAT@MCF

Catalase H2O2 decomposition by FOX assay

CAT@ZIF-8 · Powder

CAT@ZIF-8 in 6 and 8 M urea; Tris buffer pH 7.5, 50 mM.

Context: ZIF-8 enzyme-in-MOF comparison
Measurement source: S30 · Figure Section · Figure S12

Property	Reported value	Normalised value	Uncertainty	Origin and quality	Source
kobs for CAT@ZIF-8 in 6 M urea	(1.25 +/- 0.03) x 10^-3 s^-1	—	+/- 0.00003 s-1	Figure Axis Rounded Reported	S30 · Figure Section · Figure S12
kobs for CAT@ZIF-8 in 8 M urea	(8.34 +/- 0.01) x 10^-4 s^-1	—	+/- 0.000001 s-1	Figure Axis Rounded Reported	S30 · Figure Section · Figure S12

Catalase H2O2 decomposition by FOX assay

CAT@ZIF-90 · Powder

CAT@ZIF-90 incubated in Tris buffer (pH 7.5, 50 mM) with 0, 0.05, 6 or 8 M urea, 80% DMF, 0.01 M 3-AT, or 80 deg C heat treatment; H2O2 introduced at 0.1 mM.

Context: Enzyme-in-MOF target sample
Measurement source: S10 · Examinations of catalytic activities · Table S1

Property	Reported value	Normalised value	Uncertainty	Origin and quality	Source
kobs after 0.05 M urea	(2.10 +/- 0.40) x 10^-2 s^-1	—	+/- 0.0040 s-1	SI Table Exact Reported	S16 · Table Section · Table S1
kobs after 3-AT	(7.10 +/- 1.80) x 10^-6 s^-1	—	+/- 0.0000018 s-1	SI Table Exact Reported	S16 · Table Section · Table S1
kobs after 6 M ureaMarked as a best value within this paper	(1.30 +/- 0.05) x 10^-3 s^-1	—	+/- 0.00005 s-1	SI Table Exact Reported	1 · Abstract · Figure 1 / Table S1
kobs after 80 deg C treatmentMarked as a best value within this paper	(1.05 +/- 0.07) x 10^-3 s^-1	—	+/- 0.00007 s-1	SI Table Exact Reported	3 · Heat treatment · Figure 4 / Table S1
kobs after 80% DMF	(2.86 +/- 0.39) x 10^-3 s^-1	—	+/- 0.00039 s-1	SI Table Exact Reported	S16 · Table Section · Table S1
kobs after 8 M urea	(6.00 +/- 0.17) x 10^-4 s^-1	—	+/- 0.000017 s-1	SI Table Exact Reported	S16 · Table Section · Table S1
kobs, buffer without urea	(2.49 +/- 0.04) x 10^-2 s^-1	—	+/- 0.0004 s-1	SI Table Exact Reported	S16 · Table Section · Table S1

Catalase H2O2 decomposition by FOX assay

Free CAT · Unknown

Free CAT incubated in Tris buffer (pH 7.5, 50 mM) with denaturants/inhibitors; H2O2 introduced at 0.1 mM; FOX absorbance at 560 nm used for kobs.

Context: Free enzyme control
Measurement source: S10 · Examinations of catalytic activities · Table S1

Property	Reported value	Normalised value	Uncertainty	Origin and quality	Source
kobs after 0.05 M urea	(3.10 +/- 0.20) x 10^-2 s^-1	—	+/- 0.0020 s-1	SI Table Exact Reported	S16 · Table Section · Table S1
kobs after 3-AT	(1.95 +/- 0.16) x 10^-6 s^-1	—	+/- 0.00000016 s-1	SI Table Exact Reported	S16 · Table Section · Table S1
kobs after 6 M urea	(2.60 +/- 0.06) x 10^-5 s^-1	—	+/- 0.0000006 s-1	SI Table Exact Reported	S16 · Table Section · Table S1
kobs after 80 deg C treatment	(6.70 +/- 1.60) x 10^-5 s^-1	—	+/- 0.000016 s-1	SI Table Exact Reported	S16 · Table Section · Table S1
kobs after 80% DMF	(6.23 +/- 0.70) x 10^-3 s^-1	—	+/- 0.00070 s-1	SI Table Exact Reported	S16 · Table Section · Table S1
kobs after 8 M urea	(1.90 +/- 0.60) x 10^-6 s^-1	—	+/- 0.0000006 s-1	SI Table Exact Reported	S16 · Table Section · Table S1
kobs, buffer without urea	(8.90 +/- 1.7) x 10^-1 s^-1	—	+/- 0.17 s-1	SI Table Exact Reported	S16 · Table Section · Table S1

Michaelis-Menten enzyme kinetics by FOX assay

CAT@ZIF-90 · Powder

Initial rates for CAT@ZIF-90 in Tris buffer and in 0.05, 6 and 8 M urea; nonlinear fitting to y = Ax/(B+x).

Context: Target enzyme-in-MOF sample
Measurement source: S18 · Table Section · Table S3

Property	Reported value	Normalised value	Uncertainty	Origin and quality	Source
KM in 0.05 M urea	0.376 mM	—	—	SI Table Exact Reported	S18 · Table Section · Table S3
KM in 6 M urea	0.085 mM	—	—	SI Table Exact Reported	S18 · Table Section · Table S3
KM in 8 M urea	0.087 mM	—	—	SI Table Exact Reported	S18 · Table Section · Table S3
KM in Tris buffer	2.868 mM	—	—	SI Table Exact Reported	S18 · Table Section · Table S3
Vmax in 0.05 M urea	2.012 uM/s	—	—	SI Table Exact Reported	S18 · Table Section · Table S3
Vmax in 6 M urea	0.386 uM/s	—	—	SI Table Exact Reported	S18 · Table Section · Table S3
Vmax in 8 M urea	0.460 uM/s	—	—	SI Table Exact Reported	S18 · Table Section · Table S3
Vmax in Tris bufferMarked as a best value within this paper	31.068 uM/s	—	—	SI Table Exact Reported	S18 · Table Section · Table S3

Maintained activity from FOX assay

CAT@ZIF-90 · Powder

Maintained activity after incubation with 0.05 M urea or 0.1 M 3-AT, compared among free CAT, CAT@MCF and CAT@ZIF-90.

Context: Composite and free-enzyme comparison
Measurement source: 2 · Catalytic testing · Figure 2a

Property	Reported value	Normalised value	Uncertainty	Origin and quality	Source
Maintained activity after 0.05 M urea	7.1%	—	—	Figure Axis Rounded Reported	2 · Catalytic testing · Figure 2a
Maintained activity after 0.05 M ureaMarked as a best value within this paper	85.5%	—	—	Text Exact Reported	2 · Catalytic testing · Figure 2a
Maintained activity after 0.1 M 3-AT	0%	—	—	Figure Axis Rounded Reported	2 · 3-AT control · Figure 2a
Maintained activity after 0.05 M urea	3.5%	—	—	Text Exact Reported	2 · Catalytic testing · Figure 2a

Thermogravimetric analysis and Bradford assay for protein loading

Thermogravimetric analysis

CAT@ZIF-90 · Powder

TGA ramp rate 10 deg C min-1 in nitrogen; loading also determined by Bradford assay/SDS-PAGE controls.

Atmosphere: nitrogen
Context: Composite compared to free CAT and ZIF-90
Measurement source: 1 · Results · Figures S3 and S4

Property	Reported value	Normalised value	Uncertainty	Origin and quality	Source
Catalase loading in CAT@ZIF	6.0 wt %	—	—	Text Rounded Reported	1 · Results · Figures S3 and S4

Urea uptake by mass change and urease control assay

ZIF-90 microcrystals · Powder

ZIF-90 particles incubated with 6 M urea for various times; urea amount determined by mass difference; urease@ZIF-90 phenol-red assay verifies urea access.

Context: Pristine ZIF-90 host diffusion control
Measurement source: 2 · Diffusion control · Figure 2b

Property	Reported value	Normalised value	Uncertainty	Origin and quality	Source
Urea diffusion equilibrium time	after a 20 min incubation period	—	—	Text Rounded Reported	2 · Diffusion control · Figure 2b

Phenol red urease activity assay

Urease@ZIF-90 · Powder

Urease@ZIF-90 dispersed in Tris buffer pH 7.0, 10 mM with 0.5 M urea; 200 rpm shaking at 37 deg C; phenol red readout after 30 min.

Temperature: 310
Context: Diffusion-control enzyme-in-MOF sample
Measurement source: S10 · Examinations of catalytic activities · Figure S10

Property	Reported value	Normalised value	Uncertainty	Origin and quality	Source
Urease activity after Proteinase-K treatment	Urease@ZIF-90 composites treated with Proteinase-K retained urea decomposition functionality	—	—	Text Qualitative	S6 · Synthesis of Urease@ZIF-90 · Figure S10a
Urease loading	3.5 wt %	—	—	Text Rounded Reported	S6 · Synthesis of Urease@ZIF-90