Article
Biochemistry & Molecular Biology
Mattias Basle, Henry A. W. Padley, Floriane L. Martins, Gerlof Sebastiaan Winkler, Christof M. Jager, Anca Pordea
Summary: Artificial metalloenzymes were designed by inserting catalytically active metal complexes into biological scaffolds and engineering substrate binding, showing extended substrate scope. Kinetic studies revealed specific enzyme behaviors with different substrates, providing insights for further engineering of artificial metalloenzymes.
JOURNAL OF INORGANIC BIOCHEMISTRY
(2021)
Article
Chemistry, Multidisciplinary
Victor. K. K. Sharma, Kylee. J. J. Cosse, Thomas. P. P. Binder, Jeffrey. S. S. McFarlane, Alan. M. M. Allgeier
Summary: In this study, furfuryl alcohol (FOL) was biocatalytically produced from biomass-derived furfural (FAL) using ethanol (EtOH) as the terminal reductant, employing NAD(P)H-dependent yeast alcohol dehydrogenase I (YADH) as the catalyst. The conversion of FAL to FOL reached up to 74% under pH 8 with initial FAL and EtOH concentrations of 40 mM and 160 mM, respectively. This research lays the foundation for developing alternative processes to biomass-derived ethanol.
ACS SUSTAINABLE CHEMISTRY & ENGINEERING
(2023)
Article
Agriculture, Multidisciplinary
Jingyi Zhao, Yan Guo, Qiufeng Li, Jing Chen, Debao Niu, Jidong Liu
Summary: This study successfully improved the biosynthesis efficiency of allitol by increasing the intracellular NAD+ concentration, reconstructing a cofactor self-sufficient system, and utilizing a multienzyme self-assembly system.
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
(2022)
Article
Multidisciplinary Sciences
Lara Selles Vidal, James W. Murray, John T. Heap
Summary: Designing enzymes with new or improved properties is challenging, and linking the improvement in the target to cell growth through artificial selection pressure is a broadly-applicable approach. Industrial applications often require new or improved enzymes, and a semi-rational approach involving diversification of parent enzymes followed by evaluation of isolated variants can be effective.
NATURE COMMUNICATIONS
(2021)
Article
Chemistry, Physical
Miquel Garcia-Bofill, Peter W. Sutton, Marina Guillen, Gregorio Alvaro
Summary: Statins inhibit the synthesis of LDL-cholesterol related to cardiovascular diseases. ADH99 and NOX are used for the oxidation of chlorolactol to chlorolactone, leading to improved reaction yield and efficiency.
APPLIED CATALYSIS A-GENERAL
(2021)
Article
Biochemistry & Molecular Biology
Snehal D. Ganjave, Ruchika Annie O'Niel, Pramod P. Wangikar
Summary: In this study, a systematic process for refolding fungal dehydrogenases was developed to improve their availability for biocatalytic applications. Glycerol and reducing environment were found to be essential for the refolding of these enzymes, and slow dilution of solubilized protein significantly improved the recovery of refolded enzymes.
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
(2023)
Article
Biotechnology & Applied Microbiology
Yan Qiao, Can Wang, Yin Zeng, Tairan Wang, Jingjing Qiao, Chenze Lu, Zhao Wang, Xiangxian Ying
Summary: The study successfully constructed a strain expressing a fusion enzyme that achieved efficient NADP(+) regeneration and the selective oxidation of various alpha, beta-unsaturated alcohols. The researchers also optimized reaction conditions and established a platform for synthesizing valuable alpha, beta-unsaturated aldehydes.
MICROBIAL CELL FACTORIES
(2021)
Article
Microbiology
Viktor Varga, Vladimir Stefuca, Lenka Mihalova, Zdenko Levarski, Eva Struharnanska, Jaroslav Blasko, Robert Kubinec, Pavel Farkas, Vladimir Sitkey, Jan Turna, Michal Rosenberg, Stanislav Stuchlik
Summary: This study aimed to develop immobilized enzyme systems for reducing carbonyl compounds to alcohols. Recombinant enzymes were used, such as Saccharomyces cerevisiae alcohol dehydrogenase and Candida boidinii formate dehydrogenase, to convert trans-2-hexenal to trans-2-hexenol. Another system using Rhodococcus ruber alcohol dehydrogenase converted acetophenone to (S)-1-phenylethanol. All enzymes were immobilized by poly-His tag to Ni2+ for reuse after enzyme activity loss.
FRONTIERS IN MICROBIOLOGY
(2021)
Article
Biotechnology & Applied Microbiology
Xiaoshuang Wang, Fengli Wu, Dan Zhou, Guotian Song, Wujiu Chen, Cuiying Zhang, Qinhong Wang
Summary: In this study, a series of NADPH self-sufficient whole-cell biocatalysts were constructed, and the biosynthesis of shikimic acid (SA) without exogenously supplementing cofactor NADP(+) or NADPH was successfully achieved. Efficient relay-race synthesis of SA from glucose by coupling microbial fermentation with a biocatalytic process was finally demonstrated.
FERMENTATION-BASEL
(2022)
Article
Biochemistry & Molecular Biology
Karolina Bachosz, Adam Piasecki, Agata Zdarta, Ewa Kaczorek, Manuel Pinelo, Jakub Zdarta, Teofil Jesionowski
Summary: This study presents the co-immobilization of xylose dehydrogenase and alcohol dehydrogenase from Saccharomyces cerevisiae on a nanofiltration membrane for xylose bioconversion. The co-immobilized enzymes showed effectiveness and improved stability, allowing for high efficiency in the production of xylonic acid.
BIOORGANIC CHEMISTRY
(2022)
Article
Chemistry, Multidisciplinary
Willy W. L. See, Joel P. S. Choo, Do-Yun Jung, Li Zhi
Summary: Biocatalytic cascades that involve enzymatic oxidation as key steps are powerful tools for accessing valuable chemicals with various functionalizations starting from simple substrates, without the need to isolate intermediates. These oxidative cascades have diverse applications, producing a wide range of high-value products.
CURRENT OPINION IN GREEN AND SUSTAINABLE CHEMISTRY
(2022)
Article
Biochemistry & Molecular Biology
Jingjing Huo, Yajun Bai, Tai-Ping Fan, Xiaohui Zheng, Yujie Cai
Summary: In this research, a bio-catalysis cascade reaction was designed and constructed to produce hydroxytyrosol from L-3,4-dihydroxyphenylalanine. The recombinant Escherichia coli cells were used to express multiple enzymes, and glucose dehydrogenase was used to regenerate NADH, an essential reductant. The production of hydroxytyrosol reached a high yield within a short period of time under optimal conditions.
PROCESS BIOCHEMISTRY
(2022)
Article
Biochemical Research Methods
Peter Bults, Anna van der Voort, Coby Meijer, Gabe S. Sonke, Rainer Bischoff, Nico C. van de Merbel
Summary: A liquid chromatography-tandem mass spectrometry method was developed for quantifying the in vivo deamidation of biopharmaceutical proteins trastuzumab and pertuzumab. The study found significant differences between total and non-deamidated concentrations, indicating substantial deamidation in plasma. Additionally, a correlation was observed between the degree of deamidation and pharmacological activity, with deamidated trastuzumab losing its capability to inhibit the growth of breast cancer cells.
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
(2022)
Article
Chemistry, Physical
Hengyu Li, Wenhe Zhang, Changli Che, Huibin Wang, Yutian Jia, Xiao Gao, Xian Jia, Bin Qin, Song You
Summary: This study successfully switched the cofactor preference of SDR5 from NADPH to NADH through rational engineering. The key intermediates of telotristat ethyl and crizotinib were synthesized using the Mu2 variant coupled with an FDH-catalyzed cofactor regeneration system. A co-expressed biocatalyst was developed to convert ketone 1 to the desired product (R)-2.
Article
Multidisciplinary Sciences
Arsenij Kokorin, Pavel D. Parshin, Patrick J. Bakkes, Anastasia A. Pometun, Vladimir Tishkov, Vlada B. Urlacher
Summary: The fusion of multiple enzymes as multifunctional constructs has been shown to improve enzymatic properties, with noticeable increases in substrate conversion and activity observed. The enhanced diflavin reductase activity of P450 BM3 was identified as a key factor contributing to the improved performance of the fusion constructs.
SCIENTIFIC REPORTS
(2021)
Review
Biotechnology & Applied Microbiology
Tabea Bartsch, Martin Becker, Jascha Rolf, Katrin Rosenthal, Stephan Lutz
Summary: Cyclic dinucleotides (CDNs), as strong agonists of the stimulator of interferon genes, have great potential in pharmaceutical applications. The synthesis of CDNs through chemical and biocatalytic methods has its advantages and challenges. This review discusses the latest discoveries and trends in CDN synthesis, with a focus on the most promising biotechnological processes for CDN production.
BIOTECHNOLOGY AND BIOENGINEERING
(2022)
Article
Biochemistry & Molecular Biology
Jascha Rolf, Anna Christina Reyes Ngo, Stephan Lutz, Dirk Tischler, Katrin Rosenthal
Summary: A screening approach combining CFPS and colorimetric activity assay was developed to rapidly identify potential azoreductases. Ten novel azoreductases were discovered, all of which were capable of degrading methyl red dye and some could degrade other complex dyes as well. The study also revealed that NADH was the preferred electron donor for most azoreductases.
Article
Biotechnology & Applied Microbiology
Michael Osthege, Niklas Tenhaef, Rebecca Zyla, Carolin Mueller, Johannes Hemmerich, Wolfgang Wiechert, Stephan Noack, Marco Oldiges
Summary: This study introduces the Python package bletl and demonstrates its ability to perform robust data analysis and apply machine learning techniques in microbioreactor (MBR) data. The package can read raw result files from BioLector devices and utilize Python scientific computing tools for data processing and visualization. The study also proposes a new method for quantifying time-variable growth rate and showcases the application of machine learning methods for unsupervised phenotype characterization in MBR data.
ENGINEERING IN LIFE SCIENCES
(2022)
Article
Biotechnology & Applied Microbiology
Laura M. Helleckes, Carolin Muller, Tim Griesbach, Vera Waffenschmidt, Matthias Moch, Michael Osthege, Wolfgang Wiechert, Marco Oldiges
Summary: This study utilizes an automated screening process and a Bayesian statistical modeling framework to identify the optimal signal peptides for two PETases. The results from microscale cultivation are scalable to laboratory-scale bioreactors. This work demonstrates an experimental-modeling loop that improves screening efficiency and focuses on the most promising strain candidates.
BIOTECHNOLOGY AND BIOENGINEERING
(2023)
Review
Energy & Fuels
Anita Salic, Bruno Zelic
Summary: Miniaturization is a widely used approach to intensify processes, including energy production. Microstructured devices, such as microfluidic systems, offer the possibility of creating sustainable, durable, and cost-effective energy production systems based on renewable sources like biohydrogen. The challenge lies in improving the sustainability of biohydrogen production through higher substrate conversion, production rate, and yield.
Article
Chemistry, Multidisciplinary
Johannes Buchler, Eimear Hegarty, Kirsten Schroer, Radka Snajdrova, Nicholas J. J. Turner, Olivier Loiseleur, Rebecca Buller, Camille Le Chapelain
Summary: Novartis AG and Syngenta AG collaborated with the Competence Center for Biocatalysis (CCBIO) to develop a sustainable approach to late-stage halogenation using machine-learning guided protein engineering. They successfully evolved versatile catalysts capable of selectively chlorinating complex molecules, demonstrating the synthetic usefulness of such a strategy.
HELVETICA CHIMICA ACTA
(2023)
Article
Biochemistry & Molecular Biology
Sebastian Kruth, Cindy J. -M. Zimmermann, Katharina Kuhr, Wolf Hiller, Stephan Luetz, Joerg Pietruszka, Marcel Kaiser, Markus Nett
Summary: In this study, it was found that the natural product aurachin D not only has activity against the causative agent of malaria, but also inhibits other parasitic protozoa. While it has limited effects on Trypanosoma brucei rhodesiense, it shows strong killing effects on T. cruzi and Leishmania donovani at low concentrations. The IC50 values of aurachin D are even lower than the reference drugs benznidazole and miltefosine.
Article
Chemistry, Physical
Milena Mlakic, Anabela Ljubic, Anita Salic, Bruno Zelic, Otto Horvath, Valentina Milasinovic, Martin Gojun, Kresimir Molcanov, Irena Skoric
Summary: A simple and efficient protocol was utilized to transform a thiophene analog of E-resveratrol using photocatalytic oxygenation. The use of anionic and cationic free-base porphyrins, as well as their manganese(III) complexes, allowed for the formation of various photoproducts. The use of a microflow reactor significantly improved the control and yield of the photoproducts.
Article
Chemistry, Physical
Lea Winand, Stefanie Theisen, Stephan Luetz, Katrin Rosenthal, Markus Nett
Summary: The chemical synthesis of heterocycles often involves high temperature, acid or base addition, and hazardous reagents. Biocatalytic routes for heterocycle formation have gained attention due to their ability to avoid these drawbacks. In this study, the enzyme MxcM was immobilized using a hexahistidine tag, resulting in improved tolerance to organic solvents and increased stability. An immobilized enzyme reactor (IMER) was also developed, which maintained its biocatalytic activity and mechanical stability over time. These findings provide valuable insights for incorporating amidohydrolases in chemical processes for heterocycle formation.
Article
Biotechnology & Applied Microbiology
Sara Andelovic, Marko Bozinovic, Zeljka Curic, Anita Salic, Ana Jurinjak Tusek, Kristina Zagajski Kucan, Marko Rogosic, Mia Radovic, Marina Cvjetko Bubalo, Bruno Zelic
Summary: This study investigates the use of deep eutectic solvent (DES) assisted extraction for biodiesel purification and determines the optimal extraction conditions. The combination of a mathematical model and experimental results estimates the number of micro-extractors required for complete removal of glycerol.
BIOENGINEERING-BASEL
(2022)
Article
Engineering, Chemical
Regine Siedentop, Marlon Dziennus, Stephan Luetz, Katrin Rosenthal
Summary: By combining computational methods with wet-lab experiments, bottlenecks of a farnesyl pyrophosphate-producing in vitro enzyme cascade were identified. It was found that ATP and the precursor-isomerizing enzyme isopentenyl diphosphate delta-isomerase were limiting factors of the cascade's efficiency.
CHEMIE INGENIEUR TECHNIK
(2023)
Review
Chemistry, Physical
Tamara Jurina, Tea Sokac Cvetnic, Anita Salic, Maja Benkovic, Davor Valinger, Jasenka Gajdos Kljusuric, Bruno Zelic, Ana Jurinjak Tusek
Summary: In the past twenty years, there has been a significant increase in the use of microreactors in chemical and biochemical industrial processes. Microreactor systems ensure efficient process intensification due to excellent heat and mass transfer within the microchannels. Monitoring the concentrations in the microchannels is critical for understanding the physical and chemical processes in micromixers and microreactors. Therefore, there is a growing interest in performing in-line and on-line analyses of chemical and/or biochemical processes, creating opportunities for spectroscopic detection techniques in various industries.
Review
Chemistry, Physical
Tea Sokac Cvetnic, Anita Salic, Maja Benkovic, Tamara Jurina, Davor Valinger, Jasenka Gajdos Kljusuric, Bruno Zelic, Ana Jurinjak Tusek
Summary: Microreactors have become efficient tools for enzymatic reactions due to laminar fluid flow, which enables precise control, rapid mixing, and short residence time. Rapid heat and mass transfer, as well as high surface-to-volume ratios, are the main reasons for the increased reaction rates and yields in microreactors. They also eliminate mass transfer effects, allowing for more accurate determination of kinetic parameters of enzyme-catalyzed reactions.
Article
Biotechnology & Applied Microbiology
Jascha Rolf, Julian Handke, Frank Burzinski, Stephan Luetz, Katrin Rosenthal
Summary: Cell-free protein synthesis (CFPS) systems offer an alternative method for protein synthesis and screening. However, comparing the performance of different CFPS systems is challenging due to variations in reaction components. In this study, a calculation tool based on amino acid balancing was developed to assess CFPS efficiency. By applying this tool, highly efficient protein synthesis systems were identified, including well-established Escherichia coli system and less explored systems based on Vibrio natriegens or Leishmania tarentolae. This approach provides a basis for evaluating and optimizing CFPS systems.
BIOTECHNOLOGY PROGRESS
(2023)
Article
Electrochemistry
Regine Siedentop, Tobias Prenzel, Siegfried R. Waldvogel, Katrin Rosenthal, Stephan Luetz
Summary: Electrochemically coupled ATP regeneration by pyruvate oxidase and acetate kinase was successfully established for the phosphorylation of mevalonate, achieving high product yield and turnover number. The system showed promising potential and outperformed other enzymatic ATP regeneration systems.