Home Bioinformatics • Download PDF by Miguel Alcalde: Directed Enzyme Evolution: Advances and Applications

Download PDF by Miguel Alcalde: Directed Enzyme Evolution: Advances and Applications

By Miguel Alcalde

ISBN-10: 3319504118

ISBN-13: 9783319504117

ISBN-10: 3319504134

ISBN-13: 9783319504131

This ebook specializes in the most major advances in enzyme engineering which were completed via directed evolution and hybrid methods. at the twenty fifth anniversary of the invention of directed evolution, this quantity is a tribute to the pioneers of this exciting study box, and while offers a finished assessment of present examine and the country of the art.

Directed molecular evolution has turn into the main trustworthy and powerful strategy to tailor enzymes, metabolic pathways or perhaps entire microorganisms with more suitable characteristics. via mirroring the Darwinian set of rules of ordinary choice on a laboratory scale, new biomolecules of precious biotechnological curiosity can now be engineered in a fashion that surpasses the bounds of nature.

The quantity is split into sections, the 1st of which supplies an replace on contemporary winning instances of enzyme ensembles from various components of the biotechnological spectrum, together with tryptophan synthases, unspecific peroxygenases, phytases, healing enzymes, stereoselective enzymes and CO2-fixing enzymes. This part additionally presents info at the directed evolution of complete cells. the second one component of the ebook summarizes various the main appropriate tools for library construction, including the long run developments aimed toward bringing jointly directed evolution and in silico/computational enzyme layout and ancestral resurrection.

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Extra info for Directed Enzyme Evolution: Advances and Applications

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Lutz et al. 1 Asparagine-Degrading Enzymes Asparaginase II from E. coli (EcAII) is currently a key component in the treatment of childhood acute lymphoblastic leukemia (ALL). Marked under the trade name Elspar [88], EcAII hydrolyzes Asn to Asp and ammonia, which results in depletion of serum levels of Asn and triggers lymphoblast apoptosis. Consequently, overall survival rates of childhood acute lymphoblastic leukemia are relatively high at close to 90% [89], yet immunogenicity of the bacterial enzyme can cause undesirable side effects [90] and renders some patients ineligible or unable to continue treatment.

8 Nitroreductases A prominent non-nucleoside-based GDEPT system is the use of bacterial nitroreductase enzymes to activate DNA-damaging nitroaromatic prodrugs [177]. Bacterial nitroreductases are a class of flavin-dependent oxidoreductases with broad substrate specificities that have made them of interest for a number of biotechnological applications including bioremediation [178] and transgenic cell labeling and ablation [179]. Most GDEPT work has focused on the nitroreductase NfsB from E. coli and its activation of the prodrug 5-(aziridin-1-yl)-2,4-dinitrobenzamide (CB1954), which when reduced is converted to a highly cytotoxic DNA cross-linker [180].

Kinetic analysis suggested that these functional gains could largely be attributed to the enzyme’s loss of function for native 2′-deoxynucleosides while leaving kinetic parameters for NAs mostly unchanged. A rationale for the observed changes was not immediately obvious; both Asn are surface residues and distal to the active site. A subsequent crystallographic study suggested that N64D destabilizes the neighboring LID region, a conformationally flexible segment that contributes to substrate binding via hydrogen-bonding interactions with the substrates 3′-OH group [144].

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Directed Enzyme Evolution: Advances and Applications by Miguel Alcalde

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