Article
Biochemistry & Molecular Biology
Hee-Seok Lee, Jaewook Lee, Seok-Hee Lee, Hyeyeong Seo, Kwang-Geun Lee, Yooheon Park
Summary: 3-Monochloro-1,2-propanediol (3-MCPD) is a chemical compound unintentionally produced during food processing, which has been linked to reproductive toxicity, genotoxicity, neurotoxicity, and kidney toxicity. In this study, it was found that 3-MCPD inhibits muscle differentiation in C2C12 myoblasts by suppressing the expression of genetic factors related to muscle differentiation.
CHEMICO-BIOLOGICAL INTERACTIONS
(2021)
Article
Food Science & Technology
Yanjun Guo, Yuanbo Zhang, Ruijie Liu, Ming Chang, Jianhua Huang, Xiaosan Wang
Summary: This study discussed the effects of Fe3+, chloride compounds, NaCl, and water on the formation of 3-MCPD and 2-MCPD esters, as well as the concentration ratio between the two. The results showed that MgCl2 had the most significant impact on the formation of the esters, while the concentration of 3-MCPD and 2-MCPD esters in frying oil was proportional to the content of Fe3+. Additionally, the isomer ratio was strongly dependent on frying time and the addition of NaCl and water.
Article
Environmental Sciences
Yang Cai, Zongzhong Liu, Tiantian Gao, Guoyi Hu, Wenjun Yin, Yan Wang, Lingli Zhao, Dexiang Xu, Hua Wang, Tian Wei
Summary: This study evaluated the risk assessment of the emerging environmental pollutant 3-MCPD using the model organism Drosophila melanogaster. The results showed that dietary exposure to 3-MCPD caused lethality, interfered with metamorphosis and ovarian development, resulting in developmental retardance, ovarian deformity, and female fecundity disorders. Mechanistically, 3-MCPD induced redox imbalance in the ovaries, characterized by increased oxidative status and decreased antioxidant activities, which contributed to the reproductive impairments and developmental retardance.
SCIENCE OF THE TOTAL ENVIRONMENT
(2023)
Article
Agriculture, Multidisciplinary
Kazuaki Yoshinaga, Kaori Yamazaki, Toshiharu Nagai, Seiya Tanaka, Naohiro Gotoh
Summary: In this study, the effects of the deodorization process on the interconversion between 3-MCPDE and GE were examined by using 3-MCPDE or GE standards containing deuterium-labeled fatty acids. The results suggest that 3-MCPDE is converted into GE and the specific fatty acid species bound to 3-MCPDE or GE may affect their interconversion.
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
(2022)
Article
Chemistry, Physical
Eva D. Schuiten, Christoffel P. S. Badenhorst, Gottfried J. Palm, Leona Berndt, Michael Lammers, Jan Mican, David Bednar, Jiri Damborsky, Uwe T. Bornscheuer
Summary: The study identified an epoxide hydrolase called CorEH that exhibits dehalogenase activity, suggesting a rare example of a protein with dual enzymatic functions. Despite the much lower dehalogenase activity compared to epoxide hydrolase activity, the presence of multiple activities in one naturally occurring protein demonstrates significant diversity.
Article
Biotechnology & Applied Microbiology
Marcel Staar, Anett Schallmey
Summary: The immobilization of HheG D114C variants in the form of cross-linked enzyme crystals (CLECs) was found to enhance stability and reusability, making them suitable for ring opening reactions of epoxides. Among different reactor types, packed bed reactor showed the highest space-time yield of 23.9 kg(product) g(CLEC)(-1) h(-1) L-reactor volume(-1) and the highest enantiomeric excess [64%].
BIOTECHNOLOGY AND BIOENGINEERING
(2023)
Article
Chemistry, Physical
Chuanhua Zhou, Xi Chen, Tong Lv, Xu Han, Jinhui Feng, Weidong Liu, Qiaqing Wu, Dunming Zhu
Summary: In this study, a unique halohydrin dehalogenase (AbHheG) was found to have high activity and alpha/beta-regioselectivity toward the ring opening of racemic styrene oxide with cyanate but with poor stereoselectivity. By reshaping the substrate-binding site of AbHheG, a variant Y15M/N182S was obtained with excellent alpha/beta-regioselectivity and stereoselectivity. This work demonstrated that halohydrin dehalogenase is a useful biocatalyst for the synthesis of both enantiomers of 4-aryl-2-oxazolidinones from readily available racemic styrene oxides.
Article
Agricultural Engineering
Ye Rim Park, Shyam Krishna, Ok Kyung Lee, Eun Yeol Lee
Summary: This study investigated the epoxidation efficacy of different types of methanotrophs and found that type II methanotrophs are more efficient in converting alkenes to epoxides. By overexpressing epoxide hydrolases (EHs) of different origins in type II methanotrophs, 1,2-diols were successfully synthesized from epoxides. This demonstrates the potential of using methanotrophs as a microbial platform for diol production and the development of continuous bioreactors for industrial applications.
BIORESOURCE TECHNOLOGY
(2023)
Review
Toxicology
Elena Fattore, Alessia Lanno, Alberto Danieli, Simone Stefano, Alice Passoni, Alessandra Roncaglioni, Renzo Bagnati, Enrico Davoli
Summary: 3-MCPD is a chiral molecule that exists naturally as a mixture of (R)- and (S)-enantiomers. It was studied as a male antifertility drug in the 1970s, but research was discontinued due to observed side effects. However, it was later found to be a widespread contaminant in processed foods. Toxicological studies have shown that 3-MCPD primarily affects the kidney and reproductive system, followed by the neurological and immune systems. Further research is needed to understand the absorption, metabolism, and long-term toxicity of 3-MCPD esters for improved risk assessment.
ARCHIVES OF TOXICOLOGY
(2023)
Article
Chemistry, Applied
Jorge Antonio Custodio-Mendoza, Andrea Ramos, Rosa Antonia Lorenzo, Antonia M. Carro
Summary: A novel, fast, and cost-effective indirect enzymatic method was developed to accurately assess the total 3-MCPD content in canned food. The method achieved outstanding analytical results and demonstrated its reliability through specific derivatization and analysis techniques.
Article
Biotechnology & Applied Microbiology
Harshada M. Salvi, Ganapati D. Yadav
Summary: Epoxide hydrolases are enzymes widely found in nature that catalyze the hydrolysis of epoxides to diols without the need for a cofactor. A method for immobilization of epoxide hydrolase based on biomimetic mineralization has been reported, resulting in organic-inorganic hybrid nanoflowers with higher catalytic activity and stability. The nanoflowers, consisting of enzyme epoxide hydrolase and Ca2+ ions, exhibit a unique hierarchical flower-like structure and have been applied for styrene oxide hydrolysis. Factors affecting morphology, catalytic activity, and stability were studied to understand the activity recovery of the synthesized hybrid nanoflowers.
JOURNAL OF BIOTECHNOLOGY
(2021)
Article
Chemistry, Applied
Ting Xu, Qingru Zeng, Xin Liu, Shensheng Xiao, Xiaoming Jiang, Yuepeng Lu, Zhiyong Gong, Yonggui Gao, Min Fang
Summary: There is an increasing number of studies on the detection method of 3-chloro-1,2-propanediol fatty acid esters (3-MCPD esters), comparing different methods for determining 3-MCPD esters. Indirect methods have an advantage over direct methods but existing indirect methods may yield unreliable results or require long hours of alkaline methanolysis. In this study, a reliable and rapid indirect method for determining 3-MCPD esters was developed by enzymolysis to 3-MCPD through enzymatic hydrolysis, with optimized conditions. The method is simple, sensitive, and suitable for detecting 3-MCPD esters in general oils.
JOURNAL OF THE AMERICAN OIL CHEMISTS SOCIETY
(2023)
Article
Biochemistry & Molecular Biology
Liqun Zhang, Pinggu Wu, Xiaoling Xiang, Dajin Yang, Liyuan Wang, Zhengyan Hu
Summary: In this study, the physical refining process of camellia oil was simulated on a laboratory scale to reduce 3-MCPDE content. Response surface methodology was used to optimize the refining process. The optimized approach achieved a 76.9% reduction in 3-MCPDE contents, with specific parameters including degumming moisture of 2.97%, degumming temperature of 50.5°C, activated clay dosage of 2.69%, deodorizing temperature of 230°C, and deodorizing time of 90 min.
Article
Chemistry, Analytical
Yesim Tugce Yaman, Gulcin Bolat, Turkan Busra Saygin, Serdar Abaci
Summary: The study reported the fabrication of a molecularly imprinted overoxidized polypyrrole MIP(oPPy) on graphene oxide (GO) modified single-use pencil graphite electrode (PGE) for the quantitative analysis of the food processing contaminant 3-MCPD. Through optimizing experimental conditions, the research achieved superior recognition performance for 3-MCPD, indicating significant promise for future food safety control applications.
SENSORS AND ACTUATORS B-CHEMICAL
(2021)
Article
Biotechnology & Applied Microbiology
Dong Zhang, Yuqing Lei, Tingting Wang, Wenqian Lin, Xingyi Chen, Minchen Wu
Summary: This study successfully synthesized (R)-pCPED through bi-enzymatic catalysis. Under optimized reaction conditions, high concentration rac-pCSO was completely hydrolyzed within 5 hours, resulting in high yield and high enantiomeric excess of (R)-pCPED.
FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY
(2022)
Article
Chemistry, Multidisciplinary
Gerlis von Haugwitz, Kian Donnelly, Mara Di Filippo, Daniel Breite, Max Phippard, Agnes Schulze, Ren Wei, Marcus Baumann, Uwe T. Bornscheuer
Summary: Poly(vinyl alcohol) (PVA) is a water-soluble synthetic vinyl polymer with remarkable physical properties. However, its biodegradability is low and existing degradation methods have disadvantages. In this study, a modified PVA film and a PQQ-independent enzymatic cascade for the degradation of PVA are presented, which can be suitable for industrial applications.
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
(2023)
Article
Chemistry, Multidisciplinary
Yannick Branson, Simone Soeltl, Carolin Buchmann, Ren Wei, Lena Schaffert, Christoffel P. S. Badenhorst, Lukas Reisky, Gernot Jaeger, Uwe T. Bornscheuer
Summary: Enzymatic degradation and recycling are effective ways to reduce the environmental impact of plastics. However, there has been no efficient enzyme reported for the hydrolysis of polyurethanes. In this study, researchers discovered urethanases from a metagenome library constructed from soil exposed to polyurethane waste. They then developed a chemoenzymatic process using urethanase for polyurethane foam recycling.
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
(2023)
Article
Chemistry, Physical
Tobias Heinks, Nicolai Montua, Michelle Teune, Jan Liedtke, Matthias Hoehne, Uwe T. Bornscheuer, Gabriele Fischer von Mollard
Summary: This study aims to compare the stability of soluble and immobilized transaminases from different species. Immobilization of transaminases enabled their reusability for at least 10 cycles and maintained full or high activity. Upscaled kinetic resolutions demonstrated high conversion, maintained enantioselectivity, and high product yields, showing their applicability.
Article
Chemistry, Multidisciplinary
Yu-Fei Ao, Shuxin Pei, Chao Xiang, Marian J. Menke, Lin Shen, Chenghai Sun, Mark Dorr, Stefan Born, Matthias Hohne, Uwe T. Bornscheuer
Summary: We created variants of ATA from Ruegeria sp. (3FCR) with improved catalytic activity and reversed stereoselectivity, and collected a high-quality dataset. We designed a modified one-hot code to describe the effects of substrates and residues within ATAs. Finally, we built a predictor for catalytic activity and stereoselectivity, and applied it for the data-driven design of optimized variants which showed improved activity. We also demonstrated that the model can predict the catalytic activity for ATA variants of another origin by retraining with additional data.
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
(2023)
Article
Biotechnology & Applied Microbiology
Stefan Brott, Ki Hyun Nam, Francois Thomas, Theresa Dutschei, Lukas Reisky, Maike Behrens, Hanna C. C. Grimm, Gurvan Michel, Thomas Schweder, Uwe T. T. Bornscheuer
Summary: Marine algae produce complex polysaccharides that can be degraded by marine heterotrophic bacteria using carbohydrate-active enzymes. The degradation of the red algal polysaccharide porphyran involves oxidative demethylation, catalyzed by cytochrome P450 monooxygenase and its redox partners. Genes encoding zinc-dependent alcohol dehydrogenases (ADHs) were found near the genes for key enzymes in this process. The physiological role of these marine ADHs was investigated and it was found that they are required for the utilization of a specific substrate, G6Me.
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
(2023)
Article
Biochemistry & Molecular Biology
Henrik Terholsen, Kamela Myrtollari, Mirna Larva, Christina Moeller, Andreas Taden, Robert Kourist, Uwe T. Bornscheuer, Daniel Kracher
Summary: Biocatalytic decarboxylation of hydroxycinnamic acids yields phenolic styrenes, which are important precursors for antioxidants, epoxy coatings, adhesives, and other polymeric materials. This study presents two robust and sensitive assays based on photometry and fluorimetry to track decarboxylation reactions with high sensitivity while avoiding product extraction and long analysis times. The optimized assay procedures were used to measure BsPAD activity and determine the kinetic constants of the purified enzyme for different hydroxycinnamic acids, showing substrate inhibition for caffeic acid.
Article
Biochemistry & Molecular Biology
Thaleia Sakoleva, Harry P. Austin, Chrysoula Tzima, Mark Doerr, Uwe T. Bornscheuer
Summary: In this study, a novel NADPH-dependent BVMO enzyme was discovered from Halopolyspora algeriensis using SSNs, which performs oxygen insertion reactions and yields valuable products. The enzyme remains stable at temperatures between 10°C to 30°C for up to five days after purification, producing normal ester products. The substrate scope for a wide range of aliphatic ketones was investigated, and the enzyme was biochemically characterized to identify optimal reaction conditions. The best substrate, with a conversion rate of 86%, was 2-dodecanone using the purified enzyme. This novel BVMO enzyme could potentially be applied in enzymatic cascades or bioprocesses utilizing aliphatic alkanes as feedstock.
Article
Chemistry, Physical
Tobias Heinks, Luisa M. Merz, Jan Liedtke, Matthias Hoehne, Luuk M. van Langen, Uwe T. Bornscheuer, Gabriele Fischer von Mollard, Per Berglund
Summary: In this study, four different amine transaminases (ATAs) were identified to catalyze the reductive amination of 5-(hydroxymethyl)furfural (HMF) and 2,5-diformylfuran (DFF). These ATAs were successfully immobilized on glutaraldehyde-functionalized amine beads and on amine beads by site-selective binding. Immobilized ATAs showed efficient reuse in the reductive amination of HMF with alanine as co-substrate, and the ATA-Spo exhibited high stability for reuse when isopropylamine was used as an amine donor. Additionally, immobilized ATA-Spo achieved high conversion in batch syntheses and continuous-flow catalysis using alanine or isopropylamine as amine donors.
Review
Chemistry, Multidisciplinary
Thomas Bayer, Luise Haenel, Jana Husarcikova, Andreas Kunzendorf, Uwe T. T. Bornscheuer
Summary: Genetically encoded biosensor systems in living cells provide versatile, cheap, and transferable tools for detecting and quantifying a wide range of small molecules. This review focuses on state-of-the-art biosensor designs and assemblies, including transcription factor-, riboswitch-, and enzyme-coupled devices, advanced fluorescent probes, and emerging two-component systems. It also highlights (bioinformatic-assisted) strategies to address contextual issues that affect the performance of biosensors in vivo. The optimized biosensing circuits enable highly sensitive monitoring of low molecular mass chemicals and challenging physicochemical properties, with applications in various fields such as carbon dioxide fixation, industrial derivatives production, and environmental toxin detection. Furthermore, this review showcases biosensors capable of assessing platform chemical biosynthesis, enzymatic plastic waste degradation, and bioadsorption of highly toxic chemicals, offering new strategies for manufacturing, recycling, and remediation to address environmental and socioeconomic challenges.
Article
Chemistry, Multidisciplinary
Marian J. Menke, Pascal Schneider, Christoffel P. S. Badenhorst, Andreas Kunzendorf, Florian Heinz, Mark Doerr, Martin A. Hayes, Uwe T. Bornscheuer
Summary: Researchers have developed a continuous fluorescence-based high-throughput assay for methyltransferase activity, which enables the screening of suitable enzymes for enzyme-catalyzed late-stage functionalization. This assay is compatible with E. coli cell lysates and provides a reliable method to determine methyltransferase activity.
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
(2023)
Article
Biochemistry & Molecular Biology
Tobias Heinks, Simon Koopmeiners, Nicolai Montua, Norbert Sewald, Matthias Hoehne, Uwe T. Bornscheuer, Gabriele Fischer von Mollard
Summary: An enzyme cascade consisting of l-amino acid oxidase (hcLAAO4) and catalase (hCAT) was immobilized together with the (S)-selective amine transaminase (ATA-Vfl) to achieve higher reaction rates and more efficient H2O2-removal. The co-immobilized enzyme cascade showed good stability and high enantiomeric purity in preparative kinetic resolutions. However, further recycling was limited due to the instability of ATA-Vfl. The use of an engineered ATA-Vfl-8M in the immobilized enzyme cascade allowed a significant reduction in the amount of co-substrate required.
Article
Biochemistry & Molecular Biology
Yannik Brack, Chenghai Sun, Dong Yi, Uwe T. Bornscheuer
Summary: Researchers have discovered new substrate switch motifs in aromatic ammonia lyases and successfully altered the enzyme's substrate specificity. In particular, introducing previously unknown residue combinations enabled a switch from tyrosine to phenylalanine in tyrosine ammonia lyases.
Article
Chemistry, Physical
Andreas Kunzendorf, Bastian Zirpel, Lars Milke, Jakob P. Ley, Uwe T. Bornscheuer
Summary: Directed evolution of ZgOMT resulted in highly regioselective biocatalysts for the preparation of hesperetin dihydrochalcone and related compounds, providing an attractive synthesis route for para-methylation of catechol scaffolds. These enzyme variants are challenging to perform with high regioselectivity using wild-type O-methyltransferases.
Article
Chemistry, Physical
Gregory Arnal, Julien Anglade, Sabine Gavalda, Vincent Tournier, Nicolas Chabot, Uwe T. Bornscheuer, Gert Weber, Alain Marty
Summary: In recent years, enzymatic recycling of PET has become a complementary solution to current thermomechanical recycling. Promising hydrolases that depolymerize PET have been found and enhanced using protein engineering methods. However, difficulties arise from the lack of homogeneity between experimental protocols, making it hard to compare enzymes' performance. This study establishes a standardized enzymatic PET hydrolysis protocol and compares four PET hydrolases, finding that LCCICCG outperforms the others.
Review
Chemistry, Physical
In Jung Kim, Soo Rin Kim, Uwe T. Bornscheuer, Ki Hyun Nam
Summary: This review provides an overview of the importance of GH11 xylanases in bioprocessing and the key factors influencing their application. The engineering methods for achieving optimal pH shifting of GH11 xylanases in industrial applications are discussed, providing guidance for future research.
