ChemInformVolume 45, Issue 16 Isocyclic Compounds ChemInform Abstract: Enzymatic Aerobic Alkene Cleavage Catalyzed by a Mn3+-Dependent Proteinase A Homologue. Wolfgang Kroutil, Wolfgang Kroutil Dep. Chem., Karl-Franzens-Univ., A-8010 Graz, AustriaSearch for more papers by this authoret al. et al., et al. et al. Dep. Chem., Karl-Franzens-Univ., A-8010 Graz, AustriaSearch for more papers by this author Wolfgang Kroutil, Wolfgang Kroutil Dep. Chem., Karl-Franzens-Univ., A-8010 Graz, AustriaSearch for more papers by this authoret al. et al., et al. et al. Dep. Chem., Karl-Franzens-Univ., A-8010 Graz, AustriaSearch for more papers by this author First published: 03 April 2014 https://doi.org/10.1002/chin.201416087AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onEmailFacebookTwitterLinkedInRedditWechat No abstract is available for this article. References Wolfgang Kroutil, et al. et al., Enzymatic Aerobic Alkene Cleavage Catalyzed by a Mn3+-Dependent Proteinase A Homologue., ChemBioChem, 2013, 14, 2427–2430. DOI: 10.1002/cbic.201300601; 10.1002/cbic.201300601 CASWeb of Science®Google Scholar Volume45, Issue16April 22, 2014 ReferencesRelatedInformation

In the past decade, systems biology has revealed great metabolic and regulatory complexity even in seemingly simple microbial systems. Metabolic engineering aims to control this complexity in order to establish sustainable and economically viable production routes for valuable chemicals. Recent advances in systems-level data generation and modeling of cellular metabolism and regulation together with tremendous progress in synthetic biology will provide the tools to put biotechnologists on the fast track for implementing novel production processes. Great potential lies in the reduction of cellular complexity by orthogonalization of metabolic modules. Here, we review recent advances that will eventually enable metabolic engineers to predict, design, and build streamlined microbial cell factories with reduced time and effort.

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Metagenomics – the application of the genomics suit of technologies to uncultivated microorganisms – is coming of age. Sophisticated technologies are being developed and adapted to this promising genetic resource to make increasing use of the seemingly boundless molecular and functional diversity. Particular progress has been made in the areas of randomly proliferating limited‐source DNA, massively parallel sequencing without cloning, isolating specific target sequences from highly complex template mixtures, high‐throughput assay systems targeting metabolic pathways, artificial transcriptional regulators activating reporter genes to indicate enzymatic substrate conversion and cDNA cloning from extracted mRNA to directly clone actively expressed genes from a microbial consortium. However, challenges still lie ahead. Most prominently, the efficient heterologous expression of a plethora of potentially interesting enzymes from unknown source organisms is not readily achieved.

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The quaternary structure of mistletoe lectin I (MLI), a type II ribosome inactivating protein, has been determined by X-ray crystallography. A definitive molecular replacement solution was determined for MLI using the co-ordinates of the homologue ricin as a search model. MLI exists as an [AB]2 dimer with internal crystallographic two-fold symmetry. Domain I of the B chains is non-covalently associated through interactions involving three looped chains (alpha, beta, gamma) in each molecule of the dimer, forming a double trefoil structure. The ricin molecule which shares 52% sequence homology with MLI has a disulphide bridge between Cys20 and Cys39 in the alpha loop. An evolutionary mutation has replaced Cys39 with serine in MLI. This mutation appears to allow the alpha loop the flexibility required to take up its place at the dimer interface, and also suggests a rationale for why ricin does not form dimers. Measurement of retention times using FPLC gel filtration confirms that dimerisation also occurs in solution between MLI B chains with an association constant Ka = 10(6) M.