01978nas a2200229   4500008004100000022001400041245011200055210006900167260007300236300001200309490000700321520111200328653002201440653002501462653003001487653000901517653001401526100002301540700003201563700002701595856012601622       2013                        eng d  a0141-813000aBiochemical characterization of recombinant cinnamoyl CoA reductase 1 (Ll-CCRH1) from Leucaena leucocephala0 aBiochemical characterization of recombinant cinnamoyl CoA reduct  aPO BOX 211, 1000 AE AMSTERDAM, NETHERLANDSbELSEVIER SCIENCE BVcJUL  a154-1590 v583 a<p>Recombinant cinnamoyl CoA reductase 1 (Ll-CCRH1) protein from Leucaena leucocephala was overexpressed in Escherichia coli BL21 (DE3) strain and purified to apparent homogeneity. Optimum pH for forward and reverse reaction was found to be 6.5 and 7.8 respectively. The enzyme was most stable around pH 6.5 at 25 degrees C for 90 min. The enzyme showed k(cat)/k(m) for feruloyl, caffeoyl, sinapoyl, coumaroyl CoA, coniferaldehyde and sinapaldehyde as 4.6, 2.4, 2.3, 1.7, 1.9 and 1.2 (x10(6) M-1 s(-1)), respectively, indicating affinity of enzyme for feruloyl CoA over other substrates and preference of reduction reaction over oxidation. Activation energy, E-a for various substrates was found to be in the range of 20-50 kJ/mol. Involvement of probable carboxylate ion, histidine, lysine or tyrosine at the active site of enzyme was predicted by pH activity profile. SAXS studies of protein showed radius 3.04 nm and volume 49.25 nm(3) with oblate ellipsoid shape. Finally, metal ion inhibition studies revealed that Ll-CCRH1 is a metal independent enzyme. (C) 2013 Elsevier B.V. All rights reserved.</p>10aActivation energy10aCinnamoyl CoA esters10aCinnamoyl CoA reductase 110aSAXS10astability1 aSonawane, Prashant1 aVishwakarma, Rishi, Kishore1 aKhan, Bashir, Mohammad  uhttps://library.ncl.res.in/content/biochemical-characterization-recombinant-cinnamoyl-coa-reductase-1-ll-ccrh1-leucaena-0