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Indigo is a natural dye extensively used in the global textile industry. However, the conventional synthesis of indigo using toxic compounds like aniline, formaldehyde, and hydrogen cyanide has led to environmental pollution and health risks for workers. This method also faces growing economic, sustainability, and environmental challenges. To address these issues, the concept of bio-indigo or indigo biosynthesis has been proposed as an alternative to aniline-based indigo synthesis. Among various enzymes, Flavin-containing Monooxygenases (FMOs) have shown promise in achieving a high yield of bio-indigo. However, the industrialization of indigo biosynthesis still encounters several challenges. This review focuses on the historical development of indigo biosynthesis mediated by FMOs. It highlights several factors that have hindered industrialization, including the use of unsuitable chassis (Escherichia coli), the toxicity of indole, the high cost of the substrate L-tryptophan, the water-insolubility of the product indigo, the requirement of reducing reagents such as sodium dithionite, and the relatively low yield and high cost compared to chemical synthesis. Additionally, this paper summarizes various strategies to enhance the yield of indigo synthesized by FMOs, including redundant sequence deletion, semi-rational design, cheap precursor research, NADPH regeneration, large-scale fermentation, and enhancement of water solubility of indigo.
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http://dx.doi.org/10.1016/j.biotechadv.2024.108374 | DOI Listing |
Toxicol Lett
September 2025
Department of Pathology and Laboratory Medicine, Brown University, Providence, RI, USA; Center for Alternatives to Animals in Testing, Brown University, Providence, RI, USA. Electronic address:
For the in vitro determination of toxicity on target organs in the presence of physiologically relevant human metabolism, we recently developed a two-chamber liver-target organ co-culture system in a medium-throughput 96-well format. Our proof-of-concept study using human HepaRG microtissues cultured in three-dimension (3D) and AR-CALUX reporter cells demonstrated the significantly reduced testosterone (T)-mediated androgen receptor (AR) responses in the presence of human liver metabolism. The present study further increased the scientific confidence in this two-chamber co-culture system as a flexible and robust tool to capture androgen-mediated responses by incorporating alternate AR reporter cell systems as the target and examining additional androgenic compounds.
View Article and Find Full Text PDFBiomacromolecules
September 2025
Petru Poni Institute of Macromolecular Chemistry, 41A Grigore Ghica Voda Alley, Iasi 700487, Romania.
Polymer/enzyme nanoassemblies are a class of nanomaterials of increasing interest for the medical, industrial, or environmental fields. This study comprises a thorough scrutiny of the formation of nanoassemblies between laccase and a polysaccharide, chitosan, employing various experimental and theoretical methods. The binding mechanism was investigated through fluorescence quenching and molecular dynamics simulations, which identified key amino acids involved in the interaction.
View Article and Find Full Text PDFInt J Biol Macromol
September 2025
Petru Poni Institute of Macromolecular Chemistry, Iasi, Romania; Theoretical and Physical Chemistry Institute, National Hellenic Research Foundation, Athens, Greece. Electronic address:
Hybrid nanostructures from synthetic and biological macromolecules emerged as a significant research topic as they can find applications in biomedicine, catalysis, and environmental remediation. Herein, innovative nanostructures in the form of electrostatic complexes were prepared by the interaction of laccase with a hybrid graft copolymer, CHI-g-PNIPAM, obtained by grafting poly(N-isopropylacrylamide) containing carboxylate end-groups onto chitosan chains. The size, charge, morphology, and stability of the obtained nanostructures were investigated through light scattering measurements and scanning transmission electron microscopy, demonstrating the effect of the ratio between the components on the properties of the obtained co-assembled nanostructures.
View Article and Find Full Text PDFEnzyme Microb Technol
December 2025
State Key Laboratory of Food Science and Resources, Jiangnan University, Wuxi, Jiangsu 214122, China; School of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu 214122, China. Electronic address:
Indigo is an important blue pigment widely used in textile, food, and medicine industries. Biological production of indigo attracts increasing attention recently. Cell factory production of indigo encounters the problem of the toxicity of the precursor indole.
View Article and Find Full Text PDFFront Pharmacol
July 2025
Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing, China.
Background: Leukemia is a malignant hematologic disease that poses a serious threat to human health. Realgar-Indigo Naturalis formula (RIF), a traditional Chinese medicine formula, have demonstrated anti-tumor activity, but its mechanisms of action remain unclear.
Objective: This study aimed to establish a zebrafish HL-60 tumor transplantation model to investigate the anti-leukemic effects of RIF and explore its underlying mechanisms.