Titlebook: Engineering Natural Product Biosynthesis; Methods and Protocol Elizabeth Skellam Book 2022 The Editor(s) (if applicable) and The Author(s),

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Engineering Modular Polyketide Biosynthesis in , Using CRISPR/Cas: A Practical Guide,In this case, it is notably applied to rationally modify the biosynthetic pathways giving rise to the polyketide natural products, which are heavily exploited in the medical and agricultural arenas. Our aim here is to provide the potential user with a practical guide to exploit this approach for man

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CRISPR/Cas9-Based Methods for Inactivating Actinobacterial Biosynthetic Genes and Elucidating Functwe report a standard experimental protocol for the deletion of a biosynthetic gene in a . species, using the vector pCRISPomyces-2 developed by Huimin Zhao and collaborators. We also describe how carrying out metabolite analysis can reveal the putative biosynthetic function of the inactivated gene.

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Understanding and Manipulating Assembly Line Biosynthesis by Heterologous Expression in ,products. Due to the sequential biochemistry processed in each domain, the domain architecture of the assembly line enzymes strictly correlates with the product molecule. This colinearity makes assembly line enzymes an ideal target for rational reprogramming. Although many of the past engineering at

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Heterologous Expression, Purification, and Characterization of Type II Polyketide Synthase Acyl Carable cost-effective and sustainable access to a range of pharmaceutically relevant molecules. Progress toward this goal hinges on gaining ample access to materials for in vitro characterizations and structural analysis of the components of these synthases. A central component of PKSs is the acyl car

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Expression of Cyanobacterial Biosynthetic Gene Clusters in ,e clusters (BGCs) that encode cyanobacterial natural products are known, the slow growth and lack of genetic tools in the native producers hampers their modification, characterization, and large-scale production. By engineering heterologous hosts for the expression of cyanobacterial BGCs, sufficient

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