02010nas a2200229   4500008004100000022001400041245011000055210006900165260007300234300001200307490000700319520113100326653004101457653001801498653002601516100001701542700001601559700002501575700002401600700002701624856012901651       2014                        eng d  a0141-813000aLeucaena sp recombinant cinnamyl alcohol dehydrogenase: purification and physicochemical characterization0 aLeucaena sp recombinant cinnamyl alcohol dehydrogenase purificat  aPO BOX 211, 1000 AE AMSTERDAM, NETHERLANDSbELSEVIER SCIENCE BVcFEB  a254-2600 v633 a<p>Cinnamyl alcohol dehydrogenase is a broad substrate specificity enzyme catalyzing the final step in monolignol biosynthesis, leading to lignin formation in plants. Here, we report characterization of a recombinant CAD homologue (LICAD2) isolated from Leucaena leucocephala. LICAD2 is 80 kDa homodimer associated with non-covalent interactions, having substrate preference toward sinapaldehyde with K-cat/K-m of 11.6 x 10(6) (M-1 s(-1)), and a possible involvement of histidine at the active site. The enzyme remains stable up to 40 C, with the deactivation rate constant (K-d*) and half-life (t(1/2)) of 0.002 and 5 h, respectively. LICAD2 showed optimal activity at pH 6.5 and 9 for reduction and oxidation reactions, respectively, and was stable between pH 7 and 9, with the deactivation rate constant (K-d*) and half-life (t(1/2)) of 7.5 x 10(-4) and 15 h, respectively. It is a Zn-metalloenzyme with 4 Zn2+ per dimer, however, was inhibited in presence of externally supplemented Zn2+ ions. The enzyme was resistant to osmolytes, reducing agents and non-ionic detergents. (C) 2013 Elsevier B.V. All rights reserved.</p>10aCinnamyl alcohol dehydrogenase (CAD)10aMetalloenzyme10aSubstrate specificity1 aPatel, Parth1 aGupta, Neha1 aGaikwad, Sushama, M.1 aAgrawal, Dinesh, C.1 aKhan, Bashir, Mohammad  uhttps://library.ncl.res.in/content/leucaena-sp-recombinant-cinnamyl-alcohol-dehydrogenase-purification-and-physicochemical-002188nas a2200217   4500008004100000022001500041245008900056210006900145260000800214300001200222490000800234520147700242100002001719700001701739700001901756700001901775700002501794700001501819700002101834856011501855       2018                        eng d  a 0141-813000aThe Leishmania donovani IMPACT-like protein possesses non-specific nuclease activity0 aLeishmania donovani IMPACTlike protein possesses nonspecific nuc  cNOV  a962-9730 v1193 aIMPACT (Imprinted and Ancient)-like proteins are known to be regulators of GCN2 (General control non-derepressible 2) kinases involved in translation regulation. Here, we report on cloning and characterization of an IMPACT-like protein, LdIMPACT from Leishmania donovani which harbours two domains. 'RWD domain' at the N-terminal end that mediates GCN2 regulation, while a conserved 'ancient domain' lies at the C-terminal end whose function remains elusive. Interestingly, our observations indicated that LdIMPACT has a novel nonspecific nuclease activity. In silico analysis further revealed the resemblance of ancient domain of LdIMPACT to RNase PH domain (known to bind to nucleic acids). The recombinant LdIMPACT exhibited a Mg2+-dependent nuclease activity. Moreover, thermostability and pH stability assays of the protein suggest it to be a stress responsive protein. Circular dichroism studies elucidated the conformational transitions of the enzyme in response to various temperature and pH conditions which correlated well with the activity profiles. Thus, the current study highlights the structural and functional characteristics of LdIMPACT which interestingly also possesses a novel nuclease activity. With its physiological relevance unresolved, the multifaceted LdIMPACT might therefore lie in a hitherto unknown network, whose perturbation could be an attractive therapeutic approach for treating leishmaniasis. (C) 2018 Elsevier B.V. All rights reserved.1 aRao, Shilpa, J.1 aShukla, Ekta1 aBhatia, Varsha1 aLohiya, Bharat1 aGaikwad, Sushama, M.1 aKar, Anita1 aPal, Jayanta, K.  uhttps://library.ncl.res.in/content/leishmania-donovani-impact-protein-possesses-non-specific-nuclease-activity