Robin B. Kinnel

Combining Mass Spectrometric Metabolic Profiling with Genomic Analysis: A Powerful Approach for Discovering Natural Products from Cyanobacteria.

An innovative approach was developed for the discovery of new natural products by combining mass spectrometric metabolic profiling with genomic analysis and resulted in the discovery of the columbamides, a new class of di- and trichlorinated acyl amides with cannabinomimetic activity. Three species of cultured marine cyanobacteria, Moorea producens 3L, Moorea producens JHB, and Moorea bouillonii PNG, were subjected to genome sequencing and analysis for their recognizable biosynthetic pathways, and this information was then compared with their respective metabolomes as detected by MS profiling. By genome analysis, a presumed regulatory domain was identified upstream of several previously described biosynthetic gene clusters in two of these cyanobacteria, M. producens 3L and M. producens JHB. A similar regulatory domain was identified in the M. bouillonii PNG genome, and a corresponding downstream biosynthetic gene cluster was located and carefully analyzed. Subsequently, MS-based molecular networking identified a series of candidate products, and these were isolated and their structures rigorously established. On the basis of their distinctive acyl amide structure, the most prevalent metabolite was evaluated for cannabinomimetic properties and found to be moderate affinity ligands for CB1.

Kakelokelose, a sulfated mannose polysaccharide with anti-HIV activity from the Pacific tunicate Didemnum molle

An unusual sulfated mannose homopolysaccharide (1) showing in vitro anti-HIV activity has been isolated from the mucous secretion of the Pacific tunicate Didemnum molle. Analysis of 1H and 13C NMR data of the polysaccharide and its desulfated derivative 2 revealed that it consists of a sequence of 2,3-disulfated mannose units joined through β (1,6) glycosidic linkages.

Minor and trace sterols in marine invertebrates 38: isolation, structure elucidation and partial synthesis of papakusterol, a new biosynthetically unusual marine sterol with a cyclopropyl-containing side chain☆

Abstract A new cyclopropyl-containing sterol papakusterol (22-dehydro-24,26-cyclocholesterol), arising by a hitherto unknown biosynthetic process, was isolated from six “deep sea” gorgonians and its structure elucidated by 1 H-NMR analysis and partial synthesis. Traces of the corresponding Δ 5, 7 -diene have also been encountered.

Food Processing by Animals: do Beavers Leach Tree Bark to Improve Palatability?

Beavers store and consume tree parts in the bodies of water where they live. We examined whether such soaking renders food more palatable by leaching out undesirable compounds. In experiment 1, saplings of red maple, Acer rubrum (RM), were first soaked in a pond for periods of 2, 18, and 36 days, then offered to free-ranging beavers. Soaking for two days rendered RM slightly more acceptable to beavers. To further examine the time window around two days, RM sticks were soaked in distilled water in the laboratory for 1, 2, 4, and 6 days before presenting them to beavers (experiment 2). In experiment 3, twigs of three species were placed on land. Beavers placed RM in the water for 1 to 3 days before consuming the twigs. In experiment 4, sticks were provided in the water at Cranberry Lake Biological Station (CLBS). Most quaking aspen (QA) was consumed during the first night, and most witch hazel, Hamamelis virginiana (WH), during the third night. At Allegany State Park (ASP), no such difference was found. Twigs were provided in the water in experiment 5. At ASP, WH was taken after three days in the water, and at CLBS little WH was consumed, and only during the third night. A meta-analysis of all experiments shows that relatively more WH is consumed after two days than any other species. Experiment 6 traced the time beavers left their own harvested branches in the water. Unlike other tree species, WH remained in the water for two to four days before being consumed. Experiment 7 measured the phenolics leached into water from RM twigs and small pieces of bark soaked for 10 and 8 days, respectively. Shredded bark lost 50–60% of leachable phenolics into the water, and twigs 70–80%. We conclude that beavers can use water to leach undesirable compounds from their food. Although this effect was not robust, our study is the first of its kind.

A Maldiisotopic Approach to Discover Natural Products: Cryptomaldamide, a Hybrid Tripeptide from the Marine Cyanobacterium Moorea producens

Genome sequencing of microorganisms has revealed a greatly increased capacity for natural products biosynthesis than was previously recognized from compound isolation efforts alone. Hence, new methods are needed for the discovery and description of this hidden secondary metabolite potential. Here we show that provision of heavy nitrogen 15N-nitrate to marine cyanobacterial cultures followed by single-filament MALDI analysis over a period of days was highly effective in identifying a new natural product with an exceptionally high nitrogen content. The compound, named cryptomaldamide, was subsequently isolated using MS to guide the purification process, and its structure determined by 2D NMR and other spectroscopic and chromatographic methods. Bioinformatic analysis of the draft genome sequence identified a 28.7 kB gene cluster that putatively encodes for cryptomaldamide biosynthesis. Notably, an amidinotransferase is proposed to initiate the biosynthetic process by transferring an amidino group from arginine to serine to produce the first residue to be incorporated by the hybrid NRPS-PKS pathway. The maldiisotopic approach presented here is thus demonstrated to provide an orthogonal method by which to discover novel chemical diversity from Nature.