Article
Chemistry, Applied
Yosuke Sumida, Masayuki Yamasaki, Yoshiaki Nishiya, Shinya Kumagai, Toshihide Yamada, Masayuki Azuma
Summary: This study screened DSA mutants for improved enantioselectivity and thermal stability through directed evolution, with the L182E mutant showing superior performance. The crystal structure of the L182E mutant was solved, providing insight into its improved thermostability. The coupled enzymatic method involving L182E DSA and NSAR demonstrated high conversion rates and enantiomeric excesses for d-phenylalanine and d-tryptophan production, suggesting the potential industrial application of L182E DSA as a biocatalyst for d-amino acids production.
ADVANCED SYNTHESIS & CATALYSIS
(2021)
Review
Biotechnology & Applied Microbiology
Zhongdi Song, Qunfeng Zhang, Wenhui Wu, Zhongji Pu, Haoran Yu
Summary: The strategy of rational design is to predict potential mutants based on the understanding of protein structure-function relationships and introduce mutations using site-directed mutagenesis. Rational design methods are universal, fast, and have the potential for quantitative prediction of designed sequence performance. Designing enzymes with improved activity or selectivity is challenging due to the complexity of enzyme structure and limited understanding of structure-function relationships. However, with computational advancement and deeper understanding of enzyme catalysis, rational design can simplify enzyme engineering and its application is increasing.
FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY
(2023)
Article
Chemistry, Physical
Yu-Fei Ao, Hui-Juan Hu, Cheng-Xin Zhao, Peng Chen, Tingting Huang, Hui Chen, Qi-Qiang Wang, De-Xian Wang, Mei-Xiang Wang
Summary: This study demonstrates efficient biocatalytic desymmetrization of meso O-heterocyclic dicarboxamides for synthesizing both antipodes of functionalized cyclic motifs, with yields up to 99.5% ee. The engineered biocatalyst shows wide substrate promiscuity, high efficiency, and excellent enantioselectivity.
Article
Chemistry, Applied
Hui-Juan Hu, Qi-Qiang Wang, De-Xian Wang, Yu-Fei Ao
Summary: Both enantiomers of cis-1,2-disubstituted cyclohexane were obtained enantioselectively through engineered amidase-catalyzed desymmetrization of meso carbocyclic 1,2-dicarboxamides under mild conditions. The amplification and reversal of enantioselectivity of amidase was quickly achieved through generating and testing only 8 variants based on the suggested enzyme-substrate binding model by molecular docking. The synthetic application of biocatalytic meso-desymmetrization was demonstrated by straightforward transformation of the products to enantiomerically pure heterocyclic compounds.
ADVANCED SYNTHESIS & CATALYSIS
(2021)
Article
Chemistry, Physical
Ningning Zhang, Zhiyong Sun, Changzhu Wu
Summary: The polymeric approach of combining proteins with organocatalytic polymers to generate artificial enzymes expands the applications of asymmetric organocatalysis and provides new avenues for combining proteins/enzymes with well-designed organocatalytic polymers for useful synthetic reactions.
Article
Chemistry, Multidisciplinary
Changtong Zhu, Zhenbo Yuan, Zhiwei Deng, Dejing Yin, Yan Zhang, Jingwen Zhou, Yijian Rao
Summary: By engineering the enzyme GluER and combining it with photoenzymatic synthesis, the efficient asymmetric synthesis of oxygen-containing benzo-fused heterocycles has been achieved, providing a green approach for the synthesis of important chiral motifs in pharmaceuticals.
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
(2023)
Article
Chemistry, Multidisciplinary
Douglas J. Fansher, David R. J. Palmer
Summary: In this article, a biocatalytic nitro-Michael addition reaction catalyzed by NahE was reported. The reaction proceeds at room temperature with low catalyst loading, providing moderate to excellent enantioselectivity and high yields. A variety of β-nitrostyrenes reacted with pyruvate in the presence of NahE, giving β-aryl-γ-nitrobutyric acids in up to 99% yield after oxidative decarboxylation. This reaction represents the first example of an aldolase displaying promiscuous Michaelase activity and enables the use of nitroalkenes as substrates for aldolases.
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
(2023)
Article
Chemistry, Physical
Ahmed R. Aoun, Nagaraju Mupparapu, Diem N. Nguyen, Tae Ho Kim, Christopher M. Nguyen, Zhengfeiyue Pan, Sherif I. Elshahawi
Summary: The chemical properties of indole-containing molecules can be modified by functionalizing the C-H bond, and the prenylation of indole-derived compounds by indole prenyltransferases (IPTs) expands their chemical space. This study investigates the key catalytic residues that control the regiospecificity of all characterized regiospecific C6 IPTs using structure-guided site-directed mutagenesis, in vitro enzymatic reactions, kinetics, and structural elucidation of analogs. The results demonstrate that substitution of PriB_His312 to Tyr leads to the synthesis of analogs prenylated at different positions than C6. This research contributes to understanding how certain IPTs can target challenging positions in indole-derived compounds.
Article
Biochemistry & Molecular Biology
Nadim Massad, Scott Banta
Summary: Protein engineering has been utilized to enhance the activities and stabilities of enzymes by introducing synthetic swing arms and utilizing modular assembly methods. Increasing NAD(H)-PEG swing arms has been shown to improve the performance of synthetic oxidoreductases, while also enabling the prediction of K-M values for new enzymes.
Article
Chemistry, Applied
Balaji Sundara Sekar, Jiwei Mao, Benedict Ryan Lukito, Zilong Wang, Zhi Li
Summary: Enzyme cascades were developed to convert styrene into enantiopure (R)- and (S)-2-phenylglycinols, as well as other substituted styrenes and indene into substituted phenylglycinols and 1-amino-2-indanol with high enantioselectivity. Enantioselective enzyme cascades were also developed to produce (R)- and (S)-2-phenylglycinols from bio-based L-phenylalanine. Combining L-phenylalanine biosynthesis pathway with enantioselective cascade in one strain or coupled strains enabled the synthesis of (R)- and (S)-2-phenylglycinols from renewable feedstocks, enhancing production from glucose and glycerol.
ADVANCED SYNTHESIS & CATALYSIS
(2021)
Review
Biochemistry & Molecular Biology
Marek Bucko, Katarina Kaniakova, Helena Hronska, Peter Gemeiner, Michal Rosenberg
Summary: This review discusses the state of the art and development potential of epoxide hydrolases as biocatalysts, focusing on their application as chiral building blocks for bioactive compounds and drugs. The review covers new approaches to discover epoxide hydrolases using genome mining and enzyme metagenomics, as well as techniques to improve enzyme activity, enantioselectivity, enantioconvergence, and thermostability. The study also explores possibilities for expanding the synthetic capabilities of epoxide hydrolases through non-natural enzyme cascade reactions.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2023)
Review
Chemistry, Multidisciplinary
Johannes Grosskopf, Thorsten Bach
Summary: Upon irradiation with a suitable chiral catalyst, racemic compounds can be converted into enantiopure compounds through a process called photochemical deracemization. This review comprehensively covers the research and current developments in this field, discussing the reactions and mechanistic details underlying the presented reaction. Photochemical deracemization offers a unique opportunity to directly obtain enantiopure compounds from racemates, and short-lived intermediates play a key role in determining the enantioselectivity.
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
(2023)
Article
Chemistry, Physical
Matteo Capone, Gianluca Dell'Orletta, Bryce T. Nicholls, Gregory D. Scholes, Todd K. Hyster, Massimiliano Aschi, Isabella Daidone
Summary: The origin of enantioselectivity in the photoinduced radical cyclization reaction catalyzed by ene-reductases, specifically Gluconobacter oxidans ene-reductase and Old Yellow Enzyme 1, is demonstrated through molecular simulations and mutational studies. The results indicate that enantioselectivity is controlled by transition-state stability, but the relative stability of prochiral transition states is not determined by direct protein interaction, but rather by an intrinsic degree of freedom within the substrate.
Article
Chemistry, Multidisciplinary
Guanghao Huang, Regis Guillot, Cyrille Kouklovsky, Boris Maryasin, Aurelien de la Torre
Summary: A broadly applicable diastereo- and enantioselective inverse-electron-demand Diels-Alder reaction of 2-pyrones and acyclic enol ethers is reported, leading to the synthesis of bridged bicyclic lactones with high yields and enantioselectivities using a copper(II)-BOX catalytic system. Mechanistic studies suggest a stepwise mechanism, and the synthetic potential of the bridged bicyclic lactones is demonstrated through the enantioselective synthesis of polyfunctional cyclohexenes and cyclohexadienes, as well as a carbasugar unit.
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
(2022)
Article
Chemistry, Multidisciplinary
Qinglin Zhang, Kang Liang, Chang Guo
Summary: A novel electrolytic system has been introduced to divert standard ionic reactivity, providing a brand-new pattern of reactivity-electricity-driven asymmetric catalysis, which serves as a privileged chiral platform for enantioselective radical allylation.
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
(2022)
Article
Biochemistry & Molecular Biology
Douglas Weber, David Patsch, Annika Neumann, Margit Winkler, Dorte Rother
Summary: The challenge of accessing aldehydes from carboxylate moieties was overcome by utilizing carboxylate reductases (CARs) with optimized cultivation conditions, post-translational modification, and appropriate expression strain selection to successfully increase the specific activity of Nocardia otitidiscaviarum (NoCAR) by 110-fold. This highly active NoCAR could reduce sodium benzoate to benzaldehyde with high selectivity in just 2 hours.
Article
Chemistry, Physical
Tim Gerlach, David Limanhadi Nugroho, Doerte Rother
Summary: PLP-dependent enzymes are versatile biocatalysts with various industrial applications, but they can be sensitive to light conditions in laboratory settings. One variant of amine transaminases appeared to be unaffected by light exposure and even showed increased activity within 6 hours.
Editorial Material
Chemistry, Physical
Dorte Rother, Stephan Malzacher
Article
Chemistry, Analytical
Ines Reinartz, Mona Sarter, Julia Otten, Henning Hoefig, Martina Pohl, Alexander Schug, Andreas M. Stadler, Joerg Fitter
Summary: This study investigated the construction and performance of a glucose sensor equipped with fluorescent proteins by attaching different linker sequences to a glucose binding protein. The research found that the careful design of attachment of the donor fluorescent protein is crucial for the proper transfer of the glucose induced conformational change in the sensor.
Article
Chemistry, Multidisciplinary
Moritz Doeker, Laura Grabowski, Dorte Rother, Andreas Jupke
Summary: Amine transaminase catalyzed reaction systems can efficiently produce metaraminol via extraction-based in situ product removal (ISPR), leading to higher yields and improved enzymatic production. By using l-alanine as an alternative amine donor and oleic acid as a sustainable reactant, a greener production route for metaraminol was established, overcoming thermodynamic limitations and achieving higher conversions.
Article
Chemistry, Physical
Julia Hindges Nee Bramski, Johannes Doebber, Marc R. Hayes, Thomas Classen, Martina Pohl, Joerg Pietruszka
Summary: This study presents a selective synthesis method for the production of 3-hydroxyaldehydes as single aldol products of DERA. The immobilization of suitable E. coli DERA variants enabled their continuous use in synthesis. Precise reaction control using flow chemistry made the selective formation of 3-hydroxyoctanal possible. Optimization using design of experiment resulted in a 90% conversion rate. Additionally, five two-step reaction systems have been established combining biocatalysis in flow mode and consecutive chemical reactions in either continuous flow or semi-continuous flow.
Article
Chemistry, Applied
Jan -Dirk Sporing, William Graf von Westarp, Carina Ronja Kipp, Andreas Jupke, Dorte Rother
Summary: In this study, a simultaneous enzymatic two-step, one-pot cascade was developed for the synthesis of high purity vicinal diols. Different reaction systems for in situ product removal were compared, and purification processes for each system were developed. The results showed that in situ product removal in a biphasic reaction system significantly reduced the energy demand.
ORGANIC PROCESS RESEARCH & DEVELOPMENT
(2022)
Article
Chemistry, Multidisciplinary
Mohamed Labib, Laura Grabowski, Christian Bruesseler, Nicolai Kallscheuer, Luisa Wachtendonk, Thomas Fuchs, Andreas Jupke, Wolfgang Wiechert, Jan Marienhagen, Doerte Rother, Stephan Noack
Summary: This study achieved the microbial production of pyruvate and L-alanine in a one-pot fermentation process, and then transformed them into Metaraminol with high yields and stereoselectivity through a series of reactions.
ACS SUSTAINABLE CHEMISTRY & ENGINEERING
(2022)
Article
Chemistry, Physical
Selina Seide, Lilia Arnold, Solange Wetzels, Mariela Bregu, Jochem Gaetgens, Martina Pohl
Summary: This study successfully immobilized three KDOs using two immobilization techniques, with the HaloTag(R)-based immobilization achieving the best results. The stereoselective hydroxylation of L-lysine side chain was achieved, resulting in increased product titers.
Article
Engineering, Chemical
Hendrik G. Mengers, Nils Guntermann, William Graf von Westarp, Andreas Jupke, Juergen Klankermayer, Lars M. Blank, Walter Leitner, Doerte Rother
Summary: This article introduces how combinations of different catalyst types can facilitate integrated reaction sequences with biogenic substrates to form bio-hybrid fuels and products.
CHEMIE INGENIEUR TECHNIK
(2022)
Article
Chemistry, Multidisciplinary
Jan-Dirk Spoering, Jan Wiesenthal, Victoria S. Pfennig, Jochem Gaetgens, Kassem Beydoun, Carsten Bolm, Juergen Klankermayer, Doerte Rother
Summary: This study demonstrates that a complete chemoenzymatic cascade to form dioxolanes can be carried out in a purely organic environment. The highly stereoselective synthesis of the product was achieved through a two-step enzyme cascade and a ruthenium molecular catalyst.
Article
Biotechnology & Applied Microbiology
Sara Jonsdottir Glaser, Sang-Hyun Pyo, Nicola Rehnberg, Dorte Rother, Rajni Hatti-Kaul
Summary: This study evaluated the use of whole cells of Escherichia coli carrying recombinant Pseudomonas fluorescens benzaldehyde lyase as biocatalysts for carboligation of 5-HMF. The results showed that using 2 g(CDW)/L recombinant cells in 10% dimethyl carbonate, pH 8.0 at 30°C yielded 81.7% DHMF and 96.7% BHMF. The DHMF concentration reached 53.0 g/L after five feeds of 20 g/L 5-HMF.
MICROBIAL CELL FACTORIES
(2023)
Article
Chemistry, Multidisciplinary
Douglas Weber, Lucas de Souza Bastos, Margit Winkler, Yeke Ni, Abil E. Aliev, Helen C. Hailes, Doerte Rother
Summary: This study presents two multi-enzyme catalysed processes to produce 1,3,4-substituted tetrahydroisoquinoline (THIQ). The reduction of 3-hydroxybenzoic acid into 3-hydroxybenzaldehyde was catalysed by a carboxylate reductase enzyme. Two different recycling approaches, using purified enzymes or lyophilised whole-cells, were established and compared, with the whole-cell system showing superior performance.
Article
Chemistry, Physical
Reinhard Oeggl, Juliane Glaser, Eric von Lieres, Dorte Rother
Summary: The implementation of biocatalysis in flow chemistry aligns with green chemistry principles, but high substrate concentrations are often hindered by solubility issues or toxicity. Continuous synthesis of (S)-2-HPP from benzaldehyde and acetaldehyde in a micro-aqueous system requires spatial distribution of acetaldehyde to maintain sub-toxic levels.
CATALYSIS SCIENCE & TECHNOLOGY
(2021)
Article
Chemistry, Multidisciplinary
Kevin Mack, Moritz Doeker, Laura Grabowski, Andreas Jupke, Doerte Rother
Summary: The study demonstrates the effectiveness of using the bio-based amine donor l-alanine in combination with efficient process intensification through in situ product removal for biocatalytic drug synthesis.