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The upside-down jellyfish Cassiopea xamachana (Scyphozoa: Rhizostomeae) has been predominantly studied to understand its interaction with the endosymbiotic dinoflagellate algae Symbiodinium. As an easily culturable and tractable cnidarian model, it is an attractive alternative to stony corals to understanding the mechanisms driving establishment and maintenance of symbiosis. Cassiopea is also unique in requiring the symbiont in order to complete its transition to the adult stage, thereby providing an excellent model to understand symbiosis-driven development and evolution. Recently, the Cassiopea research system has gained interest beyond symbiosis in fields related to embryology, climate ecology, behavior, and more. With these developments, resources Ohdera et al. Cassiopea xamachana System Review including genomes, transcriptomes, and laboratory protocols are steadily increasing. This review provides an overview of the broad range of interdisciplinary research that has utilized the Cassiopea model and highlights the advantages of using the model for future research.

Kopi Luwak, an exotic Indonesian coffee, is made from coffee berries that have been eaten by the Asian palm civet ( Paradoxurus hermaphroditus ). Despite being known as the world's most expensive coffee, there is no reliable, standardized method for determining its authenticity. GC-MS-based multimarker profiling was employed to explore significant metabolites as discriminant markers for authentication. Extracts of 21 coffee beans ( Coffea arabica and Coffea canephora ) from three cultivation areas were analyzed and subjected to multivariate analyses, principal component analysis, and orthogonal projection to latent structures discriminant analysis. Citric acid, malic acid, and the inositol/pyroglutamic acid ratio were selected for further verification by evaluating their differentiating abilities against various commercial coffee products. The markers demonstrated potential application in the differentiation of original, fake Kopi Luwak, regular coffee, and coffee blend samples with 50 wt % Kopi Luwak content. This is the first report to address the selection and successful validation of discriminant markers for the authentication of Kopi Luwak.

A B S T R A C THigh titer 1-butanol production in Escherichia coli has previously been achieved by overexpression of a modified clostridial 1-butanol production pathway and subsequent deletion of native fermentation pathways. This strategy couples growth with production as 1-butanol pathway offers the only available terminal electron acceptors required for growth in anaerobic conditions. With further inclusion of other well-established metabolic engineering principles, a titer of 15 g/L has been obtained. In achieving this titer, many currently existing strategies have been exhausted, and 1-butanol toxicity level has been surpassed. Therefore, continued engineering of the host strain for increased production requires implementation of alternative strategies that seek to identify non-obvious targets for improvement. In this study, a metabolomics-driven approach was used to reveal a CoA imbalance resulting from a pta deletion that caused undesirable accumulation of pyruvate, butanoate, and other CoA-derived compounds. Using metabolomics, the reduction of butanoyl-CoA to butanal catalyzed by alcohol dehydrogenase AdhE2 was determined as a rate-limiting step. Fine-tuning of this activity and subsequent release of free CoA restored the CoA balance that resulted in a titer of 18.3 g/L upon improvement of total free CoA levels using cysteine supplementation. By enhancing AdhE2 activity, carbon flux was directed towards 1-butanol production and undesirable accumulation of pyruvate and butanoate was diminished. This study represents the initial report describing the improvement of 1-butanol production in E. coli by resolving CoA imbalance, which was based on metabolome analysis and rational metabolic engineering strategies.