Structure revision of spiroleucettadine, a sponge alkaloid with a bicyclic core meager in H-atoms

Our 2004 disclosure of the amino hemiketal-containing spiroleucettadine was met with keen interest by the natural products and synthetic communities. As repeated efforts to synthesize spiroleucettadine failed and questions regarding the original structure elucidation process arose, evidence mounted against the validity of the proposed structure. The low ratio of H/C in the core of spiroleucattadine complicated the original structure elucidation process. Speculation prompted a reisolation of spiroleucettadine from an untouched portion of the original Luecetta collection and a thorough analysis of analytical data. In addition, a systematic analysis of candidate structures was performed via density functional theory (DFT) calculations; a favored high scoring structure 1b was ultimately confirmed to be spiroleucettadine via X-ray analysis of crystalline spiroleucettadine and reinforced the validity of DFT calculations in structure elucidation. We present the revised structure of spiroleucettadine, a bicyclic sponge alkaloid with a scarcity of H-atoms in its core.     

Total Biosynthesis of Melleolides from Basidiomycota Fungi: Mechanistic Analysis of the Multifunctional GMC Oxidase Mld7

Mushroom terpenoids are biologically and chemically diverse fungal metabolites. Among them, melleolides are representative sesquiterpenoids with a characteristic protoilludane skeleton. In this study, we applied a recently established hot spot knock-in method to elucidate the biosynthetic pathway leading to 1α-hydroxymelleolide. The biosynthesis of the sesquiterpene core involves the cytochrome P450 catalyzing stepwise hydroxylation of the Δ⁶-protoilludene framework and a stereochemical inversion process at the C5 position catalyzed by short-chain dehydrogenase/reductase family proteins. The highlight of the biosynthesis is that the flavoprotein Mld7 catalyzes an oxidation-triggered double-bond shift accompanying dehydration and acyl-group-assisted substitution with two different nucleophiles at the C6 position to afford the Δ⁷-protoilludene derivatives, such as melleolide and armillarivin. The complex reaction mechanism was proposed by DFT calculations. Of particular importance is that product distribution is regulated by interaction with the cell membrane.     

Population study of sizes and components of self-reproducing giant multilamellar vesicles

Population analysis of a system of self-reproducing giant multilamellar vesicles (GMVs) was carried out by means of flow cytometry. The multidimensional distribution of forward light scattering (FS), side light scattering (SS), and fluorescence (FL) intensities originating from each GMV provided information about changes in a population composed of 104 vesicles. FS-FL dot plots indicated that, after the addition of the membrane precursor, the size distribution of the newly generated vesicles was nearly the same as that of the original, but the catalyst content was reduced. This result can be interpreted as evidence for the occurrence of the self-reproduction of GMVs. Moreover, the new GMVs recovered the amount of catalyst to the initial value, keeping their size distribution constant, when a solution of the catalyst was added to the new GMVs. These results are the first experimental evidence for a novel phenomenon on GMV size distribution during their self-reproducing cycle.     

Carbohydrate-appended curdlans as a new family of glycoclusters with binding properties both for a polynucleotide and lectins

Beta-1,3-glucans having carbohydrate-appendages (alpha-D-mannoside, N-acetyl-beta-D-glucosaminide and beta-lactoside) at the C6-position of every repeating unit can be readily prepared from curdlan (a linear beta-1,3-glucan) through regioselective bromination/azidation to afford 6-azido-6-deoxycurdlan followed by chemo-selective Cu(i)-catalyzed [3 + 2]-cycloaddition with various carbohydrate modules having a terminal alkyne. The resultant carbohydrate-appended curdlans can interact with polycytosine to form stable macromolecular complexes consistent with two polysaccharide strands and one polycytosine strand. Furthermore, these macromolecular complexes show strong and specific affinity toward carbohydrate-binding proteins (lectins). Therefore, one can utilize these carbohydrate-appended curdlans as a new family of glycoclusters.     

Fatty acid chemistry at the oil-water interface: self-propelled oil droplets

Fatty acids have been investigated as boundary structures to construct artificial cells due to their dynamic properties and phase transitions. Here we have explored the possibility that fatty acid systems also demonstrate movement. An oil phase was loaded with a fatty acid anhydride precursor and introduced to an aqueous fatty acid micelle solution. The oil droplets showed autonomous, sustained movement through the aqueous media. Internal convection created a positive feedback loop, and the movement of the oil droplet was sustained as convection drove fresh precursor to the surface to become hydrolyzed. As the system progressed, more surfactant was produced and some of the oil droplets transformed into supramolecular aggregates resembling multilamellar vesicles. The oil droplets also moved directionally within chemical gradients and exhibited a type of chemotaxis.     

Characterization of a mesophilic aliphatic-aromatic copolyester-degrading fungus

We isolated 5 mesophilic microorganisms that form clear zones around the colony on an opaque medium containing the aliphatic-aromatic copolyester poly(60 mol% butylene adipate-co-40 mol% butylene terephthalate) (PBAT). Among all strains, the fungal strain NKCM1712 degraded PBAT at the fastest rate (3.5 ± 0.3 μg cm⁻² h⁻¹). Genetic and morphological analyses revealed that this strain was closely related to Isaria fumosorosea (phylum Ascomycota). Mass spectroscopic analysis revealed that the degradation products were T, AB, TB, BAB, and ABT (T, terephthalic acid unit; A, adipic acid unit; B, 1,4-butanediol unit)] in the culture of the strain that used PBAT as the sole carbon source. Furthermore, the PBAT degradation ability of this strain in terms of BOD suggested that it could utilize the PBAT degradation products as growth substrates. This is the first report of a mesophilic strain that can mineralize an aliphatic-aromatic polyester into carbon dioxide on its own.     

Stability of certain minimal submanifolds in compact symmetric spaces of rank two

In [T. Kimura, M.S. Tanaka, Totally geodesic submanifold in compact symmetric spaces of rank two, Tokyo J. Math., in press], the authors obtained the global classification of the maximal totally geodesic submanifolds in compact connected irreducible symmetric spaces of rank two. In this paper, we determine their stability as minimal submanifolds in compact symmetric spaces of rank two.