In the mol-ecule of the title compound, C(12)H(13)N(3)O(3)S, the pyrrolidine ring adopts a half-chair conformation and the dihedral angle formed by the nitro group with the benzene ring is 15.18 (18)°. In the crystal, mol-ecules are linked by N-H⋯S and C-H⋯O inter-molecular hydrogen bonds into chains parallel to the c axis.
The title compound, C13H18N2O2S, adopts a cis conformation between the methyl-benzoyl and thiono groups across their thio-urea C-N bond. However, the methyl-benzoyl group and N2CS thio-urea moiety are twisted by 15.03 (3)°. In the molecule there is an N-H⋯O hydrogen bond. In the crystal, mol-ecules are linked by O-H⋯O inter-actions, generating chains extending along the c-axis direction.
In the title compound, C12H15FN2O2S, the mol-ecule adopts a cis configuration of the fluoro-benzoyl group with respect to the thiono group about their C-N bond. The dihedral angle between the fluoro-benzoyl group and the thio-urea N2CS fragment is 69.60 (11)°. An intra-molecular N-H⋯O hydrogen bond occurs. In the crystal, mol-ecules form chains along the b-axis direction via O-H⋯S and C-H⋯O hydrogen bonds.
Refluxing a mixture of 1,10-phenanthroline, (4-fluoro-phen-yl)thio-urea and cadmium(II) chloride did not produce the expected mixed-ligand complex but formed a co-crystal of the two ligands, C12H8N2·C7H7FN2S. The asymmetric unit consists of two pairs of the co-crystal mol-ecules. In each (4-fluoro-phen-yl)thio-urea mol-ecule, the planes of the N2CS thio-urea units are almost perpendicular to the corresponding fluoro-benzene rings, subtending angles of 76.53 (7) and 85.25 (7)°. In the crystal, N-H⋯N and N-H⋯S hydrogen bonds form inversion dimers from the co-crystal pairs. A weak π-π inter-action between the phenanthroline rings [centroid-centroid distance = 3.7430 (15)Å] is also observed.
In the title compound, C10H10Cl2N2OS, the mol-ecule adopts a trans-cis conformation with respect to the position of the carbonyl group and the chloro-phenyl groups relative to the thiono group across the C-N bonds. The mol-ecule is stabilized by an N-H⋯O hydrogen bond. In the crystal, mol-ecules are linked by N-H⋯S and C-H⋯O hydrogen bonds, forming zigzag chains along the b-axis direction. C-H⋯π inter-actions are also present.
The title compound, C10H9Cl2N3O3S, adopts a trans-cis conformation with respect to the position of chloropropionyl and chloronitrobenzene groups respectively, against the thiono about their C-N bonds. The conformation is stabilized by an intra-molecular N-H⋯O hydrogen bond. In the crystal, there is a short Cl⋯Cl contact with a distance of 3.386 (13) Å.
In the title compound, C(6)H(7)ClN(+)·NCS(-), the benzene ring and the protonated amine and chloro substituents are nearly planar, with a maximum deviation of 0.002 (2) Å for the N atom. In the crystal, the mol-ecules are linked by N-H⋯N and N-H⋯S hydrogen bonds into a chain along the b axis.
In the title compound, C15H11F2N3O2S, the dihedral angle between the fluoro-benzene rings is 88.43 (10)° and that between the central semithiocarbazide grouping is 47.00 (11)°. The dihedral angle between the amide group and attached fluoro-benzene ring is 50.52 (11)°; the equivalent angle between the carbonyl-thio-amide group and its attached ring is 12.98 (10)°. The major twists in the mol-ecule occur about the C-N-N-C bonds [torsion angle = -138.7 (2)°] and the Car-Car-C-N (ar = aromatic) bonds [-132.0 (2)°]. An intra-molecular N-H⋯O hydrogen bond occurs, which generates an S(6) ring. In the crystal, the mol-ecules are linked by N-H⋯O and N-H⋯S hydrogen bonds, generating (001) sheets. Weak C-H⋯O and C-H⋯F inter-actions are also observed.
Conventional refolding methods are associated with low yields due to misfolding and high aggregation rates or very dilute proteins. In this study, we describe the optimization of the conventional methods of reverse dilution and affinity chromatography for obtaining high yields of a cysteine rich recombinant glycoside hydrolase family 19 chitinase from Streptomyces griseus HUT6037 (SgChiC). SgChiC is a potential biocontrol agent and a reference enzyme in the study and development of chitinases for various applications. The overexpression of SgChiC was previously achieved by periplasmic localization from where it was extracted by osmotic shock and then purified by hydroxyapatite column chromatography. In the present study, the successful refolding and recovery of recombinant SgChiC (r-SgChiC) from inclusion bodies (IB) by reverse dilution and column chromatography methods is respectively described. Approximately 8 mg of r-SgChiC was obtained from each method with specific activities of 28 and 52 U/mg respectively. These yields are comparable to that obtained from a 1 L culture volume of the same protein isolated from the periplasmic space of E. coli BL21 (DE3) as described in previous studies. The higher yields obtained are attributed to the successful suppression of aggregation by a stepwise reduction of denaturant from high, to intermediate, and finally to low concentrations. These methods are straight forward, requiring the use of fewer refolding agents compared with previously described refolding methods. They can be applied to the refolding of other cysteine rich proteins expressed as inclusion bodies to obtain high yields of actively folded proteins. This is the first report on the recovery of actively folded SgChiC from inclusion bodies.