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

5 structured synthesis routes

Completeness describes how fully the route could be reconstructed from the main article and supporting information.

Complete recipeSource: SI

Route 1: Other

S8 · Synthesis of CAT@MCF

SolventsTris buffer (pH 7.5, 10 mM)

AdditivesCatalase enzyme solution, 0.5 mg mL-1

Atmospherenot specified

Temperature4 deg C

Time12

Work-upCollected by filtration, washed and vacuum dried.

Activationvacuum dried

Show 3 structured reagent records

Role	Reagent	Amount / concentration	Source
Other	MCF	50.0 mg	S8 · Synthesis of CAT@MCF
Additive	catalase enzyme solution	10.0 mL · 0.5 mg mL-1 in Tris buffer	S8 · Synthesis of CAT@MCF
Solvent	Tris buffer	10.0 mL enzyme solution · pH 7.5, 10 mM	S8 · Synthesis of CAT@MCF

Complete recipeSource: SI

Route 2: Other

S5 · Synthesis of CAT@ZIF-90/CAT@ZIF-8

Metal precursorsZinc nitrate hexahydrate (371.3 mg, by analogous route)

Linker precursors2-methylimidazole (2-mIm, 410.0 mg)

SolventsD.I. water, analogous to CAT@ZIF-90

AdditivesPVP (50.0 mg, by analogous route); catalase (20.0 mg, by analogous route)

Atmospherenot specified; room-temperature aqueous reaction

Temperature42 deg C dissolution by analogous route; reaction at room temperature

Timea few minutes stirring by analogous route

Work-upCollected by centrifugation at 14,000 g, washed with excess D.I. water, vacuum-dried at room temperature.

Activationvacuum-dried at room temperature

Show 5 structured reagent records

Role	Reagent	Amount / concentration	Source
Metal Source	Zinc nitrate hexahydrate	371.3 mg by analogous route	S5 · Synthesis of CAT@ZIF-90/CAT@ZIF-8
Linker	2-methylimidazole (2-mIm)	410.0 mg	S5 · Synthesis of CAT@ZIF-90/CAT@ZIF-8
Additive	polyvinylpyrrolidone (PVP)	50.0 mg by analogous route	S5 · Synthesis of CAT@ZIF-90/CAT@ZIF-8
Additive	catalase (CAT) from bovine liver	20.0 mg by analogous route	S5 · Synthesis of CAT@ZIF-90/CAT@ZIF-8
Solvent	D.I. water	analogous to CAT@ZIF-90	S5 · Synthesis of CAT@ZIF-90/CAT@ZIF-8

Complete recipeSource: SI

Route 3: Other

S5 · Synthesis of CAT@ZIF-90/CAT@ZIF-8

Metal precursorsZinc nitrate hexahydrate (371.3 mg)

Linker precursorsImidazole-2-carboxaldehyde (ICA, 480.0 mg)

SolventsD.I. water, 3.0 mL for zinc salt plus 25.0 mL for PVP/ICA/CAT solution

AdditivesPVP (50.0 mg); catalase from bovine liver (20.0 mg)

Atmospherenot specified; room-temperature aqueous reaction

Temperature42 deg C dissolution for enzyme/linker solution; reaction at room temperature

Timea few minutes stirring

Work-upCollected by centrifugation at 14,000 g, washed with excess D.I. water, vacuum-dried at room temperature.

Activationvacuum-dried at room temperature

Show 5 structured reagent records

Role	Reagent	Amount / concentration	Source
Metal Source	Zinc nitrate hexahydrate	371.3 mg	S5 · Synthesis of CAT@ZIF-90/CAT@ZIF-8
Linker	Imidazole-2-carboxaldehyde (ICA)	480.0 mg	S5 · Synthesis of CAT@ZIF-90/CAT@ZIF-8
Additive	polyvinylpyrrolidone (PVP)	50.0 mg	S5 · Synthesis of CAT@ZIF-90/CAT@ZIF-8
Additive	catalase (CAT) from bovine liver	20.0 mg	S5 · Synthesis of CAT@ZIF-90/CAT@ZIF-8
Solvent	D.I. water	3.0 mL + 25.0 mL	S5 · Synthesis of CAT@ZIF-90/CAT@ZIF-8

Complete recipeSource: SI

Route 4: Other

S8 · Synthesis of CAT@MCF

Solvents10.0 mL 37.0% HCl and 65.0 mL H2O; acetone; HCl-ethanol mixture; ethanol

AdditivesPluronic P123 (4.0 g), 1,3,5-trimethylbenzene (6.0 g), NH4F (46.0 mg), H2SO4 activation solution

Atmospherenot specified

Temperature37-40 deg C stirring; 40 deg C aging; 100 deg C treatment; 95 deg C acid activation; 60 deg C drying

Time2 h stirring; 20 h aging; 24 h at 100 deg C; 24 h acid activation; 2 h HCl-ethanol washing twice

Work-upFiltered, washed with D.I. water, dried; acid-treated, filtered, acetone washed, dried; HCl-ethanol washed twice, ethanol washed and dried.

Activation0.5 g sample treated with 500.0 mL 48.0 wt % H2SO4 at 95 deg C for 24 h

Show 6 structured reagent records

Role	Reagent	Amount / concentration	Source
Additive	Pluronic P123	4.0 g	S8 · Synthesis of CAT@MCF
Additive	1,3,5-trimethylbenzene (TMB)	6.0 g	S8 · Synthesis of CAT@MCF
Solvent	HCl/H2O acidic solution	10.0 mL of 37.0% HCl and 65.0 mL H2O	S8 · Synthesis of CAT@MCF
Other	tetraethoxysilane (TEOS)	9.2 mL	S8 · Synthesis of CAT@MCF
Additive	NH4F	46.0 mg	S8 · Synthesis of CAT@MCF
Acid	H2SO4 solution	500.0 mL for 0.5 g sample · 48.0 wt %	S8 · Synthesis of CAT@MCF

Complete recipeSource: SI

Route 5: Other

S6 · Synthesis of Urease@ZIF-90

Metal precursorsZinc nitrate hexahydrate (371.3 mg)

Linker precursorsImidazole-2-carboxaldehyde (ICA, 480.0 mg)

SolventsD.I. water, 3.0 mL for zinc salt plus 25.0 mL for ICA/PVP/urease solution

AdditivesPVP (50.0 mg); urease (8.0 mg)

Atmospherenot specified; room-temperature aqueous reaction

Temperatureroom temperature

Timea few minutes stirring

Work-upCollected by centrifugation at 14,000 g, washed with excess D.I. water, vacuum-dried at room temperature.

Activationvacuum-dried at room temperature

Show 5 structured reagent records

Role	Reagent	Amount / concentration	Source
Metal Source	Zinc nitrate hexahydrate	371.3 mg	S6 · Synthesis of Urease@ZIF-90
Linker	Imidazole-2-carboxaldehyde (ICA)	480.0 mg	S6 · Synthesis of Urease@ZIF-90
Additive	polyvinylpyrrolidone (PVP)	50.0 mg	S6 · Synthesis of Urease@ZIF-90
Additive	urease	8.0 mg	S6 · Synthesis of Urease@ZIF-90
Solvent	D.I. water	3.0 mL + 25.0 mL	S6 · Synthesis of Urease@ZIF-90