Structure, electronic properties and catalytic behaviour of an activity-enhancing CYP102A1 (P450(BM3)) variant. by Whitehouse, C, Yang, W, Yorke, J, Tufton, H, Ogilvie, L, Bell, S, Zhou, W, Bartlam, M, Rao, Z, Wong, L

“…The substrate-free crystal structure of a five-mutation directed evolution variant of CYP102A1 (P450(BM3)) with generic activity-enhancing properties ("KT2") has been determined to 1.9-Å resolution. …”

Growth-coupled enzyme engineering through manipulation of redox cofactor regeneration by Nielsen, JR, Weusthuis, RA, Huang, WE

“…These properties can be introduced using various enzyme engineering techniques, with random mutagenesis and directed evolution (DE) often being chosen when there is a lack of structural information -or mechanistic understanding- of the enzyme. …”

Development of a robust nanoscaffold as a modular vaccine platform by Bruun, T

“…In the first part of this thesis I establish a methodology for the directed evolution of SpyCatcher-VLPs. I evolve the AP205 coat protein to attempt to address limitations in solubility and yield of the SpyCatcher-AP205 platform. …”

Engineering robust enzymes via head-to-tail cyclization using spontaneous isopeptide bond formation by Schoene, C

“…To start addressing the limitations of the current approach, I developed a phage display-based strategy for the directed evolution of SpyTag/SpyCatcher reactivity. Isopeptide-mediated cyclization is a potentially powerful approach for the enhancement of protein resilience.…”

High-value wood aroma compounds via P450BM3 catalysed sesquiterpene oxidation by Wong, XJ

“…Rational design and directed evolution are the main strategies that have been applied to engineer CYP102A1 to increase its activity for the oxidation of unnatural substrates.…”

Utilising anhydride reactivity to target endogenous human proteins by Lim, SYT

“…I developed a bacterial surface display system for directed evolution of SPM, towards improving the cleavage yield and stability of the domain. …”

Development of a clean and simple biocatalysed flavin regeneration system to aid sustainable chemical synthesis by Joseph Srinivasan, S

“…With the advent of directed evolution and enzyme engineering, new reactivities previously unknown in nature are also being discovered by scientists. …”

Insight into improved thermostability of cold-adapted staphylococcal lipase by glycine to cysteine mutation by Veno, Jiivittha, Raja Abdul Rahman, Raja Noor Zaliha, Masomian, Malihe, Mohamad Ali, Mohd Shukuri, Ahmad Kamarudin, Nor Hafizah

“…Previously, the thermostability of C-terminal truncated cold-adapted lipase from Staphylococcus epidermidis AT2 (rT-M386) was markedly enhanced by directed evolution. The newly evolved mutant, G210C, demonstrated an optimal temperature shift from 25 to 45 °C and stability up to 50 °C. …”
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Peptide Variant Detection by a Living Yeast Biosensor via an Epitope-Selective Protease by Tea Crnković, Benjamin J. Bokor, Mead E. Lockwood, Virginia W. Cornish

“…With further engineering by computational modeling and directed evolution, the biosensor could likely distinguish a wide variety of peptide sequences beyond the alanine scanning carried out here. …”
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A Highly Glyphosate-Resistant EPSPS Mutant from Laboratory Evolution by Yuan Yuan, Zhengfu Zhou, Yuhua Zhan, Xiubin Ke, Yongliang Yan, Min Lin, Pengcheng Li, Shijie Jiang, Jin Wang, Wei Lu

“…These results provide more evidence for the combination of directed evolution and rational design of protein engineering.…”
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Cryptic genetic variation shapes the adaptive evolutionary potential of enzymes by Florian Baier, Nansook Hong, Gloria Yang, Anna Pabis, Charlotte M Miton, Alexandre Barrozo, Paul D Carr, Shina CL Kamerlin, Colin J Jackson, Nobuhiko Tokuriki

“…Here, we performed comparative directed evolution of four orthologous metallo-β-lactamases toward a new function and found that different starting genotypes evolved to distinct evolutionary outcomes. …”
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Applications of Nanobodies in Mycotoxin Detection by Shaowen WU, Xinlu ZOU, Jintao XU, Shijuan YAN

“…Moreover, we point out the future directions of applying rational design, directed evolution, and peptide screen and design methods to develop nanobody variants and propose potential avenues for the future development of nanobody-based mycotoxin detection technologies.…”
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Broadening the Biocatalytic Toolbox—Screening and Expression of New Unspecific Peroxygenases by Sebastian Bormann, Harald Kellner, Johanna Hermes, Robert Herzog, René Ullrich, Christiane Liers, Roland Ulber, Martin Hofrichter, Dirk Holtmann

“…This work demonstrated that directed evolution is not necessarily required to produce UPOs in <i>P. pastoris</i> at respective titers. …”
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Biocatalytic Silylation: The Condensation of Phenols and Alcohols with Triethylsilanol by Emily I. Sparkes, Chisom S. Egedeuzu, Billie Lias, Rehana Sung, Stephanie A. Caslin, S. Yasin Tabatabaei Dakhili, Peter G. Taylor, Peter Quayle, Lu Shin Wong

“…These results suggest that Silα is a potential candidate for directed evolution toward future application as a robust and selective biocatalyst for organosiloxane chemistry.…”
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A non-canonical nucleophile unlocks a new mechanistic pathway in a designed enzyme by Amy E. Hutton, Jake Foster, Rebecca Crawshaw, Florence J. Hardy, Linus O. Johannissen, Thomas M. Lister, Emilie F. Gérard, Zachary Birch-Price, Richard Obexer, Sam Hay, Anthony P. Green

“…Abstract Directed evolution of computationally designed enzymes has provided new insights into the emergence of sophisticated catalytic sites in proteins. …”
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Structure-Functional Characteristics of the Svx Protein—The Virulence Factor of the Phytopathogenic Bacterium <i>Pectobacterium atrosepticum</i> by Natalia Tendiuk, Tatiana Konnova, Olga Petrova, Elena Osipova, Timur Mukhametzyanov, Olga Makshakova, Vladimir Gorshkov

“…As such, the Svx-like proteins of phytopathogenic species form a distinct clade, pointing to the directed evolution of these proteins to provide effective interactions with plants. …”
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Identification, heterologous expression and characterization of a new unspecific peroxygenase from Marasmius fiardii PR-910 by Xin Fu, Kexin Lin, Xiaodong Zhang, Zhiyong Guo, Lixin Kang, Aitao Li

“…With its high expression level, rMfiUPO is a promising candidate as an excellent parental template for enhancing desirable traits such as increased stability and selectivity through directed evolution, thereby meeting the necessary criteria for practical application. …”
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Improved N‐acetylneuraminic acid bioproduction by optimizing pathway for reducing intermediate accumulation by Haoyu Guo, Rongzhen Tian, Chenyun Wang, Runzhi Zhao, Xueqin Lv, Long Liu, Yanfeng Liu

“…In addition, to reduce another accumulated byproduct N‐acetylmannosamine (ManNAc), the directed evolution of N‐acetylneuraminic acid synthase and the enhancement of phosphoenolpyruvate supply was implemented. …”
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Biosensor and machine learning-aided engineering of an amaryllidaceae enzyme by Simon d’Oelsnitz, Daniel J. Diaz, Wantae Kim, Daniel J. Acosta, Tyler L. Dangerfield, Mason W. Schechter, Matthew B. Minus, James R. Howard, Hannah Do, James M. Loy, Hal S. Alper, Y. Jessie Zhang, Andrew D. Ellington

“…Here, we propose an efficient biosensor-machine learning technology stack for biocatalyst development, which we apply to engineer an Amaryllidaceae enzyme in Escherichia coli. Directed evolution is used to develop a highly sensitive (EC50 = 20 μM) and specific biosensor for the key Amaryllidaceae alkaloid branchpoint 4’-O-methylnorbelladine. …”
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Applications of phage-derived RNA-based technologies in synthetic biology by Wenhui Zhang, Qiong Wu

“…Studies on phages provide rich sources of essential elements for synthetic circuit design as well as powerful support for the improvement of directed evolution platforms. Therefore, phages play a vital role in the development of new technologies and central scientific concepts. …”
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