Novel Alkaloids from Marine Actinobacteria: Discovery and Characterization

The marine environment is an excellent resource for natural products with therapeutic potential. Its microbial inhabitants, often associated with other marine organisms, are specialized in the synthesis of bioactive secondary metabolites. Similar to their terrestrial counterparts, marine Actinobacteria are a prevalent source of these natural products. Here, we discuss 77 newly discovered alkaloids produced by such marine Actinobacteria between 2017 and mid-2021, as well as the strategies employed in their elucidation. While 12 different classes of alkaloids were unraveled, indoles, diketopiperazines, glutarimides, indolizidines, and pyrroles were most dominant. Discoveries were mainly based on experimental approaches where microbial extracts were analyzed in relation to novel compounds. Although such experimental procedures have proven useful in the past, the methodologies need adaptations to limit the chance of compound rediscovery. On the other hand, genome mining provides a different angle for natural product discovery. While the technology is still relatively young compared to experimental screening, significant improvement has been made in recent years. Together with synthetic biology tools, both genome mining and extract screening provide excellent opportunities for continued drug discovery from marine Actinobacteria.     

Improved solvent extraction procedure and high-performance liquid chromatography-evaporative light-scattering detector method for analysis of polar lipids from dairy materials

A normal-phase high-performance liquid chromatography-evaporative light-scattering detector method employing dichloromethane, methanol, and acetic acid/triethylamine buffer as the mobile phase was developed for analysis of polar lipids (PLs). This method was applicable for analysis of PLs from both dairy materials and soy lecithin. All of the PLs of interest such as glycolipids, phospholipids, and sphingomyelin were well separated with a total run time of 22.5 min and without necessitating the removal of neutral lipids beforehand. Peak retention times were stable, and the method was reproducible. In this study, a modified method of using solvents for extraction of PLs from dairy matrices was also investigated. The modified method offered higher extraction efficiency, consumed less time, and in some cases saved solvent use.     

24(S)-Saringosterol Prevents Cognitive Decline in a Mouse Model for Alzheimer’s Disease

We recently found that dietary supplementation with the seaweed Sargassum fusiforme, containing the preferential LXRβ-agonist 24(S)-saringosterol, prevented memory decline and reduced amyloid-β (Aβ) deposition in an Alzheimer’s disease (AD) mouse model without inducing hepatic steatosis. Here, we examined the effects of 24(S)-saringosterol as a food additive on cognition and neuropathology in AD mice. Six-month-old male APPswePS1ΔE9 mice and wildtype C57BL/6J littermates received 24(S)-saringosterol (0.5 mg/25 g body weight/day) (APPswePS1ΔE9 n = 20; C57BL/6J n = 19) or vehicle (APPswePS1ΔE9 n = 17; C57BL/6J n = 19) for 10 weeks. Cognition was assessed using object recognition and object location tasks. Sterols were analyzed by gas chromatography/mass spectrometry, Aβ and inflammatory markers by immunohistochemistry, and gene expression by quantitative real-time PCR. Hepatic lipids were quantified after Oil-Red-O staining. Administration of 24(S)-saringosterol prevented cognitive decline in APPswePS1ΔE9 mice without affecting the Aβ plaque load. Moreover, 24(S)-saringosterol prevented the increase in the inflammatory marker Iba1 in the cortex of APPswePS1ΔE9 mice (p < 0.001). Furthermore, 24(S)-saringosterol did not affect the expression of lipid metabolism-related LXR-response genes in the hippocampus nor the hepatic neutral lipid content. Thus, administration of 24(S)-saringosterol prevented cognitive decline in APPswePS1ΔE9 mice independent of effects on Aβ load and without adverse effects on liver fat content. The anti-inflammatory effects of 24(S)-saringosterol may contribute to the prevention of cognitive decline.     

Improved performance of an intensive rotifer culture system by using a nitrifying inoculum (ABIL)

Abstract A dense nitrifying culture (ABIL) has been examined for its capacity to stimulate rotifer growth in a labscale culture system. The nitrifiers were applied in different ways. When ABIL was added directly to rotifer batch cultures, it gave rise to significantly higher population densities (factor 1.5–2.5 higher, P  < 0.05). The nitrifiers were subsequently examined for their capacity to enhance the start-up of bioreactors, commonly installed in aquaculture rearing tanks. Of the different carrier materials used in these bioreactors, i.e. CaCO₃, gravel and a PVC matrix (Bionet), CaCO₃ gave by far the best results. In a third set of experiments, effectively nitrifying bioreactor systems were connected to rotifer culture tanks and operated over a period of up to 10 days. It was demonstrated that the ABIL inoculated CaCO₃-based bioreactor allowed excellent rotifer growth reaching rotifer densities up to 5500 rotifers per mL. Moreover, a new system in which the ABIL culture was recirculated through hollow fibres was developed and demonstrated to be effective for supporting rotifer growth up to 3500 rotifers per mL. Overall, the use of the dense nitrifying culture either in seed batch cultures, conventional bioreactors or hollow fibre bioreactor systems in support of rotifer cultures was demonstrated to be effective for improving the water quality and the rotifer growth.     

Quantitation of bovine immunoglobulin G2 by a competitive fluorescence immunoassay

Bovine immunoglobulin G2 (IgG2) was quantitated by fluoroimmunoassay (FIA) based on competition with fluorescein-conjugated IgG2 for Sepharose-bound anti-IgG2 antibodies. The optimal range was 0.1 to 1 microgram IgG2. The FIA was compared with the single radial immunodiffusion (SRD) test. Both methods gave comparable means and standard errors for IgG2 in serum. The FIA method was 30 times more sensitive.