Synthesis of Hybrid Cyclopeptides through Enzymatic Macrocyclization

Acknowledgements We thank Dr. G. Mann and Dr. A. Bent for supplying the enzymes and the useful discussions, and Dr. T. Lebl for the useful NMR discussions. This work was supported by the European Research Council (339367), UK Biotechnology and Biological Sciences Research Council (K015508/1), the Wellcome Trust [TripleTOF 5600 mass spectrometer (094476), the MALDI TOF-TOF Analyzer (079272AIA), 700 NMR], and the EPSRC UK National Mass Spectrometry Facility at Swansea University. J.H.N., M.A.J., and N.J.W. are named on patents that have been filed by St. Andrews and Aberdeen Universities to commercialize the enzymes (PatG) and (LynD) used in the study. No income derives from the patent.Peer reviewedPublisher PD

Bypassing the proline/thiazoline requirement of the macrocyclase PatG

Peer reviewedPublisher PD

Characterization of the fast and promiscuous macrocyclase from plant PCY1 enables the use of simple substrates

H.L. is funded by the George and Stella Lee Scholarship and EPSRC. This project was funded by the European Research Council project 339367 NCB-TNT and by the BBSRC (J.H.N.). E.S.M. and M.A. are funded by EPSRC. S.A.S. is funded by BSRC mass spec facility.Cyclic ribosomally derived peptides possess diverse bioactivities and are currently of major interest in drug development. However, it can be chemically challenging to synthesize these molecules, hindering the diversification and testing of cyclic peptide leads. Enzymes used in vitro offer a solution to this; however peptide macrocyclization remains the bottleneck. PCY1, involved in the biosynthesis of plant orbitides, belongs to the class of prolyl oligopeptidases and natively displays substrate promiscuity. PCY1 is a promising candidate for in vitro utilization, but its substrates require an 11 to 16 residue C-terminal recognition tail. We have characterized PCY1 both kinetically and structurally with multiple substrate complexes revealing the molecular basis of recognition and catalysis. Using these insights, we have identified a three residue C-terminal extension that replaces the natural recognition tail permitting PCY1 to operate on synthetic substrates. We demonstrate that PCY1 can macrocyclize a variety of substrates with this short tail, including unnatural amino acids and nonamino acids, highlighting PCY1’s potential in biocatalysis.PostprintPeer reviewe

Characterization of the fast and promiscuous macrocyclase from plant PCY1 enables the use of simple substrates

Cyclic ribosomally derived peptides possess diverse bioactivities and are currently of major interest in drug development. However, it can be chemically challenging to synthesize these molecules, hindering the diversification and testing of cyclic peptide leads. Enzymes used in vitro offer a solution to this; however peptide macrocyclization remains the bottleneck. PCY1, involved in the biosynthesis of plant orbitides, belongs to the class of prolyl oligopeptidases and natively displays substrate promiscuity. PCY1 is a promising candidate for in vitro utilization, but its substrates require an 11 to 16 residue C-terminal recognition tail. We have characterized PCY1 both kinetically and structurally with multiple substrate complexes revealing the molecular basis of recognition and catalysis. Using these insights, we have identified a three residue C-terminal extension that replaces the natural recognition tail permitting PCY1 to operate on synthetic substrates. We demonstrate that PCY1 can macrocyclize a variety of substrates with this short tail, including unnatural amino acids and nonamino acids, highlighting PCY1's potential in biocatalysis

Green communication for next-generation wireless systems: optimization strategies, challenges, solutions, and future aspects

Wireless sensor networks (WSNs) have emerged as a backbone technology for the wireless communication era. The demand for WSN is rapidly increasing due to their major role in various applications with a wider deployment and omnipresent nature. The WSN is rapidly integrated into a large number of applications such as industrial, security, monitoring, tracking, and applications in home automation. The widespread use in many different areas attracts research interest in WSNs. Therefore, researchers are taking initiatives in exploring innovation day by day particularly towards the Internet of Things (IoT). But, WSN is having lots of challenging issues that need to be addressed, and the inherent characteristics of WSN severely affect the performance. Energy constraints are one of the primary issues that require urgent attention from the research community. Optimal energy optimization strategies are needed to counter the issue of energy constraints. Although one of the most appropriate schemes for handling energy constraints issues is the appropriate energy harvesting technique, the optimal energy optimization strategies should be coupled together for effectively utilizing the harvested energy. In this high-level systematic and taxonomical survey, we have organized the energy optimization strategies for EH-WSNs into eleven factors, namely, radio optimization schemes, optimizing the energy harvesting process, data reduction schemes, schemes based on cross-layer optimization, schemes based on cross-layer optimization, sleep/wake-up policies, schemes based on load balancing, schemes based on optimization of power requirement, optimization of communication mechanism, schemes based on optimization of battery operations, mobility-based schemes, and finally energy balancing schemes. We have also prepared the summarized view of various protocols/algorithms with their remarkable details. This systematic and taxonomy survey also provides a progressive detailed overview and classification of various optimization challenges for the EH-WSNs that require attention from the researcher followed by a survey of corresponding solutions for corresponding optimization issues. Further, this systematic and taxonomical survey also provides a deep analysis of various emerging energy harvesting technologies in the last twenty years of the era