Reactions between halogen substituted terpyridine ligands F- (L¹), Cl- (L²), Br- (L³) and I-4′-phenyl-terpyridine (L⁴) and ZnBr₂ or ZnI₂ led to the formation of complexes [Zn(Br)₂L¹] (1), [Zn(I)₂L¹] (2), [Zn(Br)₂L²] (3), [Zn(I)₂L²] (4), [Zn(Br)₂L³] (5), [Zn(I)₂L³] (6), [Zn(Br)₂L⁴] (7), and [Zn(I)₂L⁴] (8), respectively, which were characterized by elemental analysis, ¹H NMR, IR and single crystal X-ray diffraction. Their photoluminescence properties, solution stabilities, hydrophilicity/lipophilicity properties and anti-proliferative activity against three cancer cell lines as well as structure–function relationships are analyzed. All the complexes, which show high solution stabilities and lipophilicity under determination conditions, reveal much higher antiproliferative activities than cisplatin against the human lung carcinoma cell line (A549), human hepatocellular carcinoma cell line (Bel-7042) and human breast cancer cell line (MCF-7) in vitro. Fluorescence spectroscopy and circular dichroic analysis reveal that the compounds have strong affinity binding with ctDNA and stack onto the base pairs of the ctDNA. The results obtained are further explained in terms of the electronic properties of the compounds by density functional theory (DFT) calculations, and it is found that the halogen anions play critical roles in the antitumor activity of these compounds. Their affinities to HSP90 proteins, ALK, EGFR and HER2 kinases and the binding sites on these proteins have been determined by computational docking analysis, and the results indicate that the only form of their binding is van der Waals forces.

In order to better utilize chitinolytic enzymes to produce high-value N-acetyl-D-glucosamine (GlcNAc) from chitinous waste, there is an urgent need to explore bi-functional chitinases with exceptional properties of temperature, pH and metal tolerance. In this study, we cloned and characterized a novel bi-functional cold-adaptive chitinase called CaChi18A from a newly isolated strain, Chitinilyticum aquatile CSC-1, in Bama longevity village of Guangxi Province, China. The activity of CaChi18A at 50 °C was 4.07 U/mg. However, it exhibited significant catalytic activity even at 5 °C. Its truncated variant CaChi18A_ΔChBDs, containing only catalytic domain, demonstrated significant activity levels, exceeding 40 %, over a temperature range of 5-60 °C and a pH range of 3 to 10. It was noteworthy that it displayed tolerance towards most metal ions at a final concentration of 0.1 mM, including Fe³⁺ and Cu²⁺ ions, retaining 122.52 ± 0.17 % and 116.42 ± 1.52 % activity, respectively. Additionally, it exhibited favorable tolerance towards organic solvents with the exception of formic acid. Interestedly, CaChi18A and CaChi18A_ΔChBDs had a low K_m value towards colloidal chitin (CC), 0.94 mg mL^-1 and 2.13 mg mL^-1, respectively. Both enzymes exhibited chitobiosidase and N-acetyl-D-glucosaminidase activities, producing GlcNAc as the primary product when hydrolyzing CC. The high activities across a broader temperature and pH range, strong environmental adaptability, and hydrolytic properties of CaChi18A_ΔChBDs suggested that it could be a promising candidate for GlcNAc production.

A series of ZnCl₂ complexes (compounds 1–10) with 4′-(substituted-phenyl)-2,2′:6′,2′′-terpyridine that bears hydrogen (L¹), p-methyl (L²), p-methoxy (L³), p-phenyl (L⁴), p-tolyl (L⁵), p-hydroxyl (L⁶), m-hydroxyl (L⁷), o-hydroxyl (L⁸), p-carboxyl (L⁹), or p-methylsulfonyl (L¹⁰) were prepared and then characterized by ¹H NMR, electrospray mass-spectra (ESI-MS), IR, elemental analysis, and single crystal X-ray diffraction. In vitro cytotoxicity assay was used to monitor the antiproliferative activities against tumor cells. Absorption spectroscopy, fluorescence titration, circular dichroism spectroscopy, and molecular modeling studied the DNA interactions. All of the compounds display interesting photoluminescent properties and different maximal emission peaks due to the difference of the substituent groups. The cell viability studies indicate that the compounds have excellent antiproliferative activity against four human carcinoma cell lines, A549, Bel-7402, MCF-7, and Eca-109, with the lowest IC₅₀ values of 0.33 (10), 0.66 (6), 0.37 (7), and 1.05 (7) μM, respectively. The spectrophotometric results reveal that the compounds have strong affinity binding with DNA as intercalator and induce DNA conformational transition. Molecular docking studies indicate that the binding is contributed by the π…π stacking and hydrogen bonds, providing an order of nucleotide sequence binding selectivity as ATGC > ATAT > GCGC. These compounds intercalate into the base pairs of the DNA of the tumor cells to affect their replication and transcription, and the process is supposed to play an important role in the anticancer mechanism.

Eleven copper chloride coordination compounds (1–11) with 4′-(4′-substituted-phenyl)-2,2′:6′,2′′-terpyridine ligands bearing hydrogen (L¹), cyano (L²), p-hydroxyl (L³), m-hydroxyl (L⁴), o-hydroxyl (L⁵), methoxyl (L⁶), iodo (L⁷), bromo (L⁸), chloro (L⁹), fluoro (L¹⁰) or methylsulfonyl (L¹¹) were prepared and characterized by IR spectroscopy, elemental analysis and single crystal X-ray diffraction. Antiproliferative activities against tumor cells were investigated and DNA interactions were studied by circular dichroism spectroscopy and molecular modeling methods. In vitro data demonstrate that all the compounds exhibit higher antiproliferative activities as compared to cisplatin against five human carcinoma cell lines: A549, Bel-7402, Eca-109, HeLa and MCF-7. Compound 6 with methoxyl shows the best anti-proliferation activity. Spectrophotometric results reveal the strong affinity of the compounds for binding with DNA as intercalators and induce DNA conformational transitions. The results of molecular docking studies show that the compounds interact with DNA through π–π stacking, van der Waals forces, hydrophobic interactions and hydrogen bonds. The binding energies between compound 11 and three macromolecules, including DNA duplex, oligonucleotide and DNA–Topo I complex, are the lowest. The binding stability of compounds containing hydroxyl, methoxy and methylsulfonyl groups with biological macromolecules mainly relies on the hydrogen bonds. The ability of a compound to form hydrogen bonds can promote its binding to biological targets, thereby exhibiting high antiproliferative activity.

In order to discover a broad-specificity and high stability chitinase, a marine fungus, Aspergillus fumigatus df347, was identified in the sediments of mangrove wetlands in Qinzhou Bay, China. The chitinase gene (AfChi28) from A. fumigatus df347 was cloned and heterologously expressed in Escherichia coli, and the recombinant enzyme AfChi28 was purified and characterized. AfChi28 is an acido-halotolerant- and temperature-resistant bifunctional enzyme with both endo- and exo-cleavage functions. Its enzymatic products are mainly GlcNAc, (GlcNAc)₂, (GlcNAc)₃ and (GlcNAc)₄. Na⁺, Mg²⁺, K⁺, Ca²⁺ and Tris at a concentration of 50 mM had a strong stimulatory effect on AfChi28. The crude enzyme and pure enzyme exhibited the highest specific activity of 0.737 mU/mg and 52.414 mU/mg towards colloidal chitin. The DxDxE motif at the end of strand β5 and with Glu154 as the catalytic residue was verified by the AlphaFold2 prediction and sequence alignment of homologous proteins. Moreover, the results of molecular docking showed that molecular modeling of chitohexaose was shown to bind to AfChi28 in subsites −4 to +2 in the deep groove substrate-binding pocket. This study demonstrates that AfChi28 is a promising chitinase for the preparation of desirable chitin oligosaccharides, and provides a foundation for elucidating the catalytic mechanism of chitinases from marine fungi.