Article
Biochemistry & Molecular Biology
Jorick Franceus, Zorica Ubiparip, Koen Beerens, Tom Desmet
Summary: 2-O-Glucosylglycerol is a substance accumulated by bacteria and plants in response to environmental stress, widely used in skin care products. In this study, a thermostable phosphorylase was engineered to improve its efficiency and stability for producing 2-O-Glucosylglycerol, resulting in a variant with increased catalytic efficiency, improved regioselectivity, and retained activity at elevated temperatures.
Article
Chemistry, Physical
Tong Zhang, Pi Liu, Hongli Wei, Xinming Sun, Yan Zeng, Xuewen Zhang, Yi Cai, Mengfei Cui, Hongwu Ma, Weidong Liu, Yuanxia Sun, Jiangang Yang
Summary: This study discovered a glucosylglycerol phosphorylase enzyme from Marinobacter adhaerens that exhibited broad catalytic properties in glycosylation of polyols, particularly high carbon 1,2-diols, with excellent regioselectivity and stereoselectivity. The researchers also improved the thermostability and catalytic efficiency of the enzyme and developed an ATP-free multienzyme system for polyol glycosylation using inexpensive starch as a donor. This efficient synthetic platform has the potential to manufacture other valuable chiral 1,2-cis-glycosides.
Article
Agricultural Engineering
Yanzhe Shang, Ping Zhang, Wenping Wei, Jin Li, Bang-Ce Ye
Summary: By optimizing carbon metabolism, increasing gene copies, and blocking degradation, a high yield of polydatin was successfully produced in Y. lipolytica, demonstrating its great potential for glycoside synthesis.
BIORESOURCE TECHNOLOGY
(2023)
Article
Chemistry, Multidisciplinary
Chenghai Sun, Bao-Di Ma, Guangjun Li, Wenya Tian, Lu Yang, Haidong Peng, Zhi Lin, Zixin Deng, Xu-Dong Kong, Xudong Qu
Summary: Through genome mining and study of the sequence-product relationship, we identified three key residues (F387, F388, and E73) that play pivotal roles in selecting different diketopiperazine (DKP) substrates. Engineering these residues in Nas(F5053) greatly expanded its substrate specificity, enabling the biosynthesis of 12 self-dimerized and at least 81 cross-dimerized HTDKPs. Structural and molecular dynamics analysis of F387G and E73S revealed that they control substrate specificity by reducing steric hindrance and regulating substrate tunnels.
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
(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
Anja Knorrscheidt, Jordi Soler, Nicole Huenecke, Pascal Puellmann, Marc Garcia-Borras, Martin J. Weissenborn
Summary: The study successfully improved the oxyfunctionalization activity of MthUPO through protein engineering, resulting in variants with high chemo- and regioselectivities. By reshaping the active site, the catalytic efficiency for model substrates was increased. This research represents a step towards the use of UPOs in biocatalytic synthetic pathways of industrial interest.
Article
Chemistry, Physical
Bernd A. Nebel, Michael Breuer, Andreas Schneider, Benjamin Aberle, Stephan C. Hammer, Per-Olof Syren, Martin J. Weissenborn, Bettina M. Nestl
Summary: This article reflects on Professor Bernhard Hauer's distinguished career in biocatalysis and highlights his contributions to the application and development of enzymes.
Article
Biochemistry & Molecular Biology
Valentina Pirillo, Marco Orlando, Davide Tessaro, Loredano Pollegioni, Gianluca Molla
Summary: This study reports a straightforward workflow for the evolution of PET-hydrolysing enzymes (PHEs) using semi-rational protein engineering and high-throughput screening. By screening variant libraries on PET nanoparticles, a variant with higher hydrolytic activity was identified and further stabilized. This workflow provides a suitable protocol for the generation of efficient enzymatic tools for polyester degradation.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2022)
Article
Biotechnology & Applied Microbiology
Alexander Sigg, Mario Klimacek, Bernd Nidetzky
Summary: This study demonstrates the use of (mechanistic-empirical) hybrid modeling for optimizing the sucrose phosphorylase-catalyzed glycosylation of glycerol to synthesize the cosmetic ingredient GG. The hybrid model includes microscopic terms of the reverse reaction to account for the full-time course of GG synthesis, enabling comprehensive window-of-operation analysis and constrained optimization of GG production. The hybrid model is shown to be a powerful tool for process decision making and rational selection of enzyme catalysts.
BIOTECHNOLOGY AND BIOENGINEERING
(2021)
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
Yongzhen Peng, Zhiguo Wang, Yang Chen, Weihua Xu, Yujing Hu, Zhichun Chen, Jian Xu, Qi Wu
Summary: In this study, a promiscuous cyclohexanone monooxygenase (CHMO) was used for the photoinduced synthesis of chiral alpha-fluoroketones. A structure-guided rational design of CHMO improved the yield and stereoselectivity of the synthesis remarkably. This methodology allows for the high-yield and enantioselective synthesis of desirable chiral alpha-fluoroketones.
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
(2022)
Article
Chemistry, Multidisciplinary
Xinyue Wang, Yijie Sheng, Haiyang Cui, Jie Qiao, Yibo Song, Xiujuan Li, He Huang
Summary: This study developed a protein engineering strategy called corner engineering to tailor enzyme resistance towards deep eutectic solvents (DESs) and high temperatures. The results demonstrated the generality of this strategy and identified the key factors influencing enzyme resistance and thermostability.
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
(2023)
Article
Chemistry, Multidisciplinary
Dengke Gao, Wei Song, Jing Wu, Liang Guo, Cong Gao, Jia Liu, Xiulai Chen, Liming Liu
Summary: In this study, an enzymatic-spontaneous chemical cascade route for the production of L-HPA was designed and successfully produced high-purity L-HPA using simple and cost-effective starting materials.
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
(2022)
Article
Chemistry, Physical
Moritz Voss, Sean Hueppi, Daniela Schaub, Takahiro Hayashi, Mathieu Ligibel, Emine Sager, Kirsten Schroer, Radka Snajdrova, Rebecca Buller
Summary: Enzymatic late-stage diversification of small molecules is a rapid method for generating diversity in compound libraries for drug discovery. The Fe(II)/alpha-ketoglutarate-dependent halogenase enzyme family has become of interest for its ability to selectively halogenate unactivated C(sp(3))-H bonds. In this study, two engineered variants of the halogenase WelO5* were developed for the racemic resolution of a mixture of stereoisomers generated during the synthesis of a bioactive martinelline-derived fragment.
Article
Biotechnology & Applied Microbiology
Christian Rapp, Stevie van Overtveldt, Koen Beerens, Hansjorg Weber, Tom Desmet, Bernd Nidetzky
Summary: Epimerization of sugar nucleotides is crucial for the diversity in monosaccharide building blocks in cellular biosynthesis. A newly discovered CDP-tyvelose 2-epimerase-like enzyme from Thermodesulfatator atlanticus shows promiscuity in promoting C-2 epimerization in various nucleotide-activated forms of D-glucose. The enzyme exhibits optimal activity at high temperature and alkaline pH, expanding the toolbox for epimerase-catalyzed reactions in sugar nucleotide synthesis.
APPLIED AND ENVIRONMENTAL MICROBIOLOGY
(2021)
Article
Chemistry, Applied
Jose L. Gonzalez-Alfonso, Zorica Ubiparip, Elena Jimenez-Ortega, Ana Poveda, Cristina Alonso, Luisa Coderch, Jesus Jimenez-Barbero, Julia Sanz-Aparicio, Antonio O. Ballesteros, Tom Desmet, Francisco J. Plou
Summary: Glycosylation of polyphenols can enhance their solubility, stability, and bioavailability. The generated phloretin glucosides showed improved solubility and moderate loss in antioxidant activity, which could be recovered upon deglycosylation. In a pig skin model, the diglucoside was mainly detected in superficial layers, indicating potential applications in dermocosmetics.
ADVANCED SYNTHESIS & CATALYSIS
(2021)
Review
Biotechnology & Applied Microbiology
Zorica Ubiparip, Marc De Doncker, Koen Beerens, Jorick Franceus, Tom Desmet
Summary: Beta-glucan phosphorylases are important carbohydrate-active enzymes with great potential for the biocatalytic synthesis of beta-glucans. Their preference for sugar phosphates highlights their significance in the carbohydrate industry.
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
(2021)
Article
Agriculture, Dairy & Animal Science
N. Van Noten, E. Van Liefferinge, J. Degroote, S. De Smet, T. Desmet, J. Michiels
Summary: Thymol alpha-D-glucopyranoside and thymol beta-D-glucopyranoside have different stability and absorption mechanisms in the porcine gut. Bacterial hydrolysis is the main cause of thymol beta-D-glucopyranoside loss, while thymol alpha-D-glucopyranoside is more resistant to degradation or absorption.
Article
Biochemistry & Molecular Biology
Emma De Beul, Alana Jongbloet, Jorick Franceus, Tom Desmet
Summary: The study focuses on the sequence diversity of Glycoside Hydrolase Family 65 (GH65) and identified specificity-determining positions through in-silico analysis of correlated mutations. By searching for amino acid motifs that differ from previously characterized enzymes, several new clades with potential new functions were identified. Three new enzymes were expressed in E. coli, with one enzyme showing high specificity in hydrolyzing specific substrates.
Article
Nutrition & Dietetics
Amar van Laar, Charlotte Grootaert, Filip Van Nieuwerburgh, Dieter Deforce, Tom Desmet, Koen Beerens, John Van Camp
Summary: The study found that different sugars have structure-specific effects on cellular energy production, hepatic fat accumulation, and gene expression, suggesting a potential health benefits of trehalose and L-arabinose beyond the distinction of conventional and rare sugars.
Review
Biochemistry & Molecular Biology
Koen Beerens, Ophelia Gevaert, Tom Desmet
Summary: This review provides a comprehensive overview of GDP-mannose 3,5-epimerase, including its catalytic complexity, biological functions, as well as its potential industrial applications in rare sugar synthesis and glycorandomization.
FRONTIERS IN MOLECULAR BIOSCIENCES
(2022)
Article
Agriculture, Multidisciplinary
Shari Dhaene, Amar Van Laar, Marc De Doncker, Emma De Beul, Koen Beerens, Charlotte Grootaert, Jurgen Caroen, Johan Van der Eycken, John Van Camp, Tom Desmet
Summary: With the global pandemic of obesity and related metabolic diseases, there is a growing interest in alternative carbohydrates as a potential replacement for traditional sugars. However, there is limited knowledge about the majority of alternative rare carbohydrates and their properties. In this study, three novel glycosides were enzymatically produced and their digestibility and caloric potential were assessed. The results showed that these rare disaccharides had reduced digestibility and limited impact on energy metabolism, indicating a low-caloric potential.
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
(2022)
Review
Biochemistry & Molecular Biology
Ulrike Vogel, Koen Beerens, Tom Desmet
Summary: Nucleotide sugar dehydratases are central to the biosynthesis of deoxy and amino sugars, which have important biological functions in all domains of life. Bacteria are capable of producing a wide range of highly specialized monosaccharides. These sugars are involved in the virulence of pathogenic species and the biosynthesis of diverse macrolide antibiotics. The activity of NS dehydratases plays a key role in the biosynthesis of deoxy sugars, and they can be classified into three groups based on their structure and reaction mechanism.
JOURNAL OF BIOLOGICAL CHEMISTRY
(2022)
Article
Biochemical Research Methods
Ulrike Vogel, Koen Beerens, Tom Desmet
Summary: Nucleotide sugar 4,6-dehydratases are crucial enzymes in the biosynthesis of deoxy and amino sugars. This study presents a colorimetric assay for the detection of their reaction products, which allows for enzyme characterization and screening in microtiter plates.
ANALYTICAL BIOCHEMISTRY
(2022)
Review
Biotechnology & Applied Microbiology
Simone Savino, Tom Desmet, Jorick Franceus
Summary: This review highlights the importance and relevance of insertions and deletions (indels) in protein evolution and engineering. The authors provide a comprehensive overview of approaches to include indels in protein engineering pipelines and discuss the tolerance to indels at the structural level as well as the potential of frameshift mutations.
BIOTECHNOLOGY ADVANCES
(2022)
Review
Biochemical Research Methods
Jorick Franceus, Jolien Lormans, Tom Desmet
Summary: Protein engineering methods have been successful in expanding the glycosylation reactions of carbohydrate-active enzymes, which is important for the commercial value of specialty carbohydrates and glycosylated compounds. Recent studies have shown how protein engineers have improved the fitness landscape of CAZymes by connecting different activity types through mutational bridges.
CURRENT OPINION IN BIOTECHNOLOGY
(2022)
Article
Nutrition & Dietetics
Amar van Laar, Charlotte Grootaert, Andreja Rajkovic, Tom Desmet, Koen Beerens, John Van Camp
Summary: Rare sugars have potential as sugar replacers and their impact on metabolism and insulin sensitivity was investigated. Glucose and fructose increased acidification rate and impaired glucose uptake in certain cells, while rare disaccharides had delayed digestion and minimal effect on insulin-stimulated glucose uptake. Further research is needed to confirm their beneficial impact on insulin sensitivity in humans.
Article
Biotechnology & Applied Microbiology
Carlos Alvarez Quispe, Matthieu Da Costa, Koen Beerens, Tom Desmet
Summary: This study identifies a novel subgroup of Gal4E enzymes with unprecedented specificity for GDP sugars. Biochemical investigation of Gal4E from Pyrococcus horikoshii reveals its clear preference for GDP sugars and its ability to accept various other sugar moieties, including L-sugars.
CURRENT RESEARCH IN BIOTECHNOLOGY
(2022)
Article
Chemistry, Multidisciplinary
Silke Geurs, Dorien Clarisse, Freya Baele, Jorick Franceus, Tom Desmet, Karolien De Bosscher, Matthias D'hooghe
Summary: In this study, a set of HDAC6-selective inhibitors with low side effects and significant inhibitory activity in the nanomolar range were successfully synthesized using a lean inhibitor strategy.
CHEMICAL COMMUNICATIONS
(2022)
