Natural products are prolific producers of diverse chemical scaffolds, which have yielded several clinically useful drugs. However, the complex features of natural products present challenges for identifying bioactive molecules using high-throughput screens. For most assays, measured endpoints are either colorimetric or luminescence based. Thus, the presence of the major metabolites, tannins, and chlorophylls, in natural products could potentially interfere with these measurements to give either false-positive or false-negative hits. In this context, zebrafish phenotypic assays provide an alternative approach to bioprospect naturally occurring bioactive compounds. Whether tannins and/or chlorophylls interfere in zebrafish phenotypic assays, is unclear. In this study, we evaluated the interference potential of tannins and chlorophylls against efficacy of known small-molecule inhibitors that are known to cause phenotypic abnormalities in developing zebrafish embryos. First, we fractionated tannin-enriched fraction (TEF) and chlorophyll-enriched fraction (CEF) from Camellia sinensis and cotreated them with PD0325901 [mitogen-activated protein kinase-kinase (MEK) inhibitor] and sunitinib malate (SM; anti-[lymph]angiogenic drug). While TEF and CEF did not interfere with phenotypic or molecular endpoints of PD0325901, TEF at 100 μg/mL partially masked the antiangiogenic effect of SM. On the other hand, CEF (100 μg/mL) was toxic when treated up to 6 dpf. Furthermore, CEF at 100 μg/mL potentially enhanced the activity of γ-secretase inhibitors, resulting in toxicity of treated embryos. Our study provides evidence that the presence of tannin and/or chlorophyll in natural products do interfere with zebrafish phenotype assays used for identifying potential hits. However, this may be target/assay dependent and thus requiring additional optimization steps to assess interference potential of tannins and chlorophylls before performing any screening assay.
The escalation of the global population has accelerated the demand for sustainable energy sources such as bioethanol. Traditionally, bioethanol was obtained by the fermentation of sugar from agricultural crops and grains. However, this technique creates serious threats on the global food supplies, thus hindering the commercial production of bioethanol. Therefore, there is a need to develop new technologies and low-cost raw materials in order to ensure that bioethanol is economically comparable to the first generation of bioethanol. Solid-state fermentation (SSF) has been in the limelight within the scientific community because of its efficiency, cost-effectiveness, and promising technology to produce bioethanol. SSF involves the cultivation of microorganisms on a solid substrate in the absence of free-flowing water, which eliminates the need for sugar extraction and reduces wastewater production. This systematic review provides an overview of the applications of SSF in bioethanol production while presenting recent studies and advancements of this technology for producing sustainable and cost-effective bioethanol.