Optimizing the formation of in vitro sponge primmorphs from the Chinese sponge Stylotella agminata (Ridley)

The establishment and optimization of in vitro primmorph formation from a Chinese sponge, Stylotella agminata (Ridley), collected from the South China Sea, were investigated. Our aims were to identify the key factors affecting primmorph formation in this species and to optimize the technique for developing an in vitro primmorph culture system. The size of dissociated cells from S. agminata is relatively small, in the range between 5 and 10 mum. Round-shaped primmorphs of less than 100 gm were formed 3 days after transferring the dissociated cells into seawater containing Ca2+ and Mg2+. The effect of various cell dissociation conditions, inoculum. cell density, concentration of antibiotics, pH, and temperature was further investigated upon the formation of primmorphs. The time required for primmorph formation, primmorph size distribution, and the proliferating capability were microscopically documented. Healthy sponge S. agminata, inoculum. cell density and culture temperature play a critical role for the successful formation of primmorphs and that the microbial contamination will have to be controlled. (C) 2002 Elsevier Science B.V. All rights reserved.

Advanced precursors in marine biosynthetic study. Part 3: The biosynthesis of dichloroimines in the tropical marine sponge Stylotella aurantium

The biosynthetic origins of the dichloroimine group in the stylotellanes A and B 1,2 have been investigated by incorporation of [C-14]-labeled farnesyl isocyanide 7 and farnesyl isothiocyanate 3 into the sponge Stylotella aurantium. (C) 2002 Elsevier Science Ltd. All rights reserled.

Biosynthetic pathways to dichloroimines; precursor incorporation studies on terpene metabolites in the tropical marine sponge Stylotella aurantium

The biosynthetic origin of the dichloroimine functional group in the marine sponge terpene metabolites stylotellanes A ( 3) and B ( 4) was probed by the use of [C-14]-labelled precursor experiments. Incubation of the sponge Stylotella aurantium with [C-14]-labelled cyanide or thiocyanate resulted in radioactive terpenes in which the radiolabel was shown by hydrolytic chemical degradation to be associated specifically with the dichloroimine carbons. Additionally, label from both precursors was incorporated into farnesyl isothiocyanate ( 2). A time course experiment with [ 14C]cyanide revealed that the specific activity for farnesyl isothiocyanate decreases over time, but increases for stylotellane B ( 4), consistent with the rapid formation of farnesyl isothiocyanate ( 2) from inorganic precursors followed by a slower conversion to stylotellane B ( 4). The advanced precursors farnesyl isothiocyanate ( 2) and farnesyl isocyanide ( 5) were supplied to S. aurantium, and shown to be incorporated efficiently into stylotellane A ( 3) and B ( 4). Feeding of [C-14]-farnesyl isothiocyanate ( 2) resulted in a higher incorporation of label than with [C-14]-farnesyl isocyanide ( 5). Farnesyl isocyanide was incorporated into farnesyl isothiocyanate in agreement with labelling studies in other marine sponges. Both farnesyl isocyanide and isothiocyanate were further incorporated into axinyssamide A ( 11) as well as the cyclized dichloroimines (12)-(14), ( 16) that represent more advanced biosynthetic products of this pathway. These results identify the likely biosynthetic pathway leading to the major metabolites of S. aurantium.

Marine isocyanides and related natural products structure, biosynthesis and ecology

This review highlights structural and biosynthetic work on a group of nitrogen-functionalised terpenes that are almost exclusively found in marine invertebrates and the animals that feed on them. The chemical functionality reviewed includes isocyanides, isothiocyanates, formamides, thiocyanates, isocyanates, and dichloroimines. The literature through mid 2003 is reviewed and there are 143 citations.