Enzymatic reductions with dihydrogen via metal catalyzed cofactor regeneration
A cofactor, metal technology used in the field of enzymatic reduction with hydrogen through metal-catalyzed cofactor regeneration
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Embodiment 1
[0112] Example 1 shows that in the presence of complexes containing ruthenium, TBADH and NADP + Under the conditions of the catalyst, H 2 Can be used to convert 2-butanone to 2-butanol. NADP + (28.0 mg, 36.6 μmol) was placed in 2 ml of 0.1 M phosphate buffer, and the pH value was adjusted to 8.1 with NaOH. To this solution was added TBADH (1.2 mg, 8.8 units) and placed in an 80 ml Fisher-Porter bottle. Evacuate the Fisher-Porter bottle and refill it with argon three times, then add [Cl 2 Ru(TPPTS) 2 ] 2 (6.4 mg, 5.0 μmol) and 2-butanone (5 μl). Evacuate the Fisher-Porter bottle and fill with 70psi of H 2 , and then heated to 40°C for two hours. The gas was released from the bottle, and GLC analysis of a liquid sample indicated the production of 9.5 μmol of 2-butanol.
Embodiment 2
[0117]Example 2 shows that in complexes containing ruthenium, TBADH and NADP + In the presence of catalyst, H 2 2-Heptanone can be converted to 2-heptanol with remarkable symmetric isomerism.
[0118] Under argon flow, fill 20ml of 0.1M phosphate buffer (pH7.0), 2-heptanone (100μl, 718μmol), TPPTS (74mg, 130.7μmol), NADP + (10 mg, 13.1 μmol) and TBADH (2.7 mg, 19.8 units). Finally add the ruthenium catalyst [Cl 2 Ru(TPPTS) 2 ] 2 (17mg, 6.5μmol) and sealed Fisher-Porter bottle, evacuated, filled with 70psiH 2 . Repeat the evacuation and filling of H twice 2 , and then heat the above apparatus to 60°C with an oil bath. GC analysis of a trace sample taken by the liquid sampler described above after 10 hours indicated the production of 227.8 μmol of 2-heptanol. An additional 13.0 μmol (240.8 μmol) had accumulated after 4 hours. The pressure was released from the above bottle and 2-heptanol was 70.5% (S) and 29.5% (R) (41.0% ee) by chiral GC. Cycle number of catalyst co...
Embodiment 3
[0120] Example 3 shows that the ruthenium catalyst [Cl 2 Ru(TPPTS) 2 ] 2 is through H 2 Restore NAD(P) + catalyst precursors.
[0121] NADP + (36.0mg, 47.0μmol) was placed in 2ml of 0.1M phosphate buffer. The pH of the solution was adjusted to 8.3 with NaOH and placed in an 80 ml Fisher-Porter bottle. Evacuate the Fisher-Porter bottle and refill it with argon three times, then add [Cl 2 Ru(TPPTS) 2 ] 2 (9.7 mg, 3.7 μmol). Evacuate the Fisher-Porter bottle and fill with 70psi of H 2 , and then heated to 40°C for two hours. The volatile gases were removed under reduced pressure, and the sample was dissolved in D 2 O middle. Add 20 μl of trimethylsilyl propionic acid standard solution to the above sample. 1 H NMR integration indicated that 29.0 μmol of NADPH was produced under these conditions. NADP + 1 H(D 2 O): δ9.35(s)N 2 , 9.17(d, 0.6Hz)N 6 , 8.84(d,1.0)N 4 , 8.42(s)A 2 , 8.21(t, 0.7)
[0122] N 5 , 8.11(s)A 8 , 6.10(d, 5.2Hz)N 1’ , 6.02(d,)A 1...
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