Article
Chemistry, Applied
Jesse T. C. Brown, Ryan M. Phelan
Summary: Chiral amines are important in the synthesis of pharmaceutical and agrochemical compounds, and amine transaminases have emerged as efficient and stereoselective catalysts. However, the compatibility between the transaminase and amine donors is a concern. In this study, we compare different amine donors and identify those that are compatible with various amine transaminases, providing insights into the preferences of engineered and wild-type enzymes.
ORGANIC PROCESS RESEARCH & DEVELOPMENT
(2023)
Article
Chemistry, Multidisciplinary
Ludwig L. Bengel, Benjamin Aberle, Alexander-N. Egler-Kemmerer, Samuel Kienzle, Bernhard Hauer, Stephan C. Hammer
Summary: Selective alkylation of pyrazoles using a catalyst-controlled enzymatic system allows for the efficient synthesis of important molecules with unprecedented regioselectivity. The use of a computational enzyme-library design tool enabled the conversion of a promiscuous enzyme into a small enzyme family of pyrazole-alkylating enzymes in one round of mutagenesis and screening. This enzymatic system demonstrates high selectivity and achieves different alkylations of pyrazoles with remarkable regiodivergence.
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
(2021)
Article
Biotechnology & Applied Microbiology
Hannah S. Zurier, Julie M. Goddard
Summary: The consequences of plastic waste affect all forms of life in both terrestrial and aquatic ecosystems. However, microbes have adapted to utilize plastic as a source of nutrients. PETase is a depolymerase enzyme that breaks down aromatic polyesters. This study developed a higher-throughput PETase engineering platform to optimize its industrial application. Using this platform, a more stable and efficient PETase variant was developed, promising advancements in the industrial depolymerization of plastic waste.
BIOTECHNOLOGY AND BIOENGINEERING
(2023)
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)
Article
Chemistry, Physical
Lea R. Rapp, Sergio M. Marques, Bernd Nebel, Jiri Damborsky, Bernhard Hauer
Summary: The catalytic space of the P450 monooxygenase CYP153A(M.aq) was transformed from a fatty acid hydroxylase to a catalyst capable of omega-hydroxylation of dodecylamine. A rapid screening method called RAPP was established to measure saturation libraries directly from a 96-deepwell plate. Molecular modeling and dynamics simulations revealed significant effects of the mutations on the substrate tunnel architectures.
Article
Biochemistry & Molecular Biology
Katja F. Hellendahl, Maryke Fehlau, Sebastian Hans, Peter Neubauer, Anke Kurreck
Summary: The study introduces a luciferase-based assay for the detection of NK activity, which was validated in a 96-well plate format. The high-throughput assay showed comparable results to HPLC analysis.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2021)
Article
Biotechnology & Applied Microbiology
Jinshan Gao, Muhua Du, Jinhua Zhao, Jinhua Zhao, Ning Xu, Huanmin Du, Jiansong Ju, Liang Wei, Jun Liu
Summary: The article introduces a genetically encoded biosensor for detecting L-cysteine levels and screening high-producing strains. By coupling with fluorescence-activated cell sorting, a high-throughput screening platform was established, which successfully improved the catalytic performance of the key enzyme in the L-cysteine biosynthetic pathway.
METABOLIC ENGINEERING
(2022)
Review
Chemistry, Multidisciplinary
Emily Radley, John Davidson, Jake Foster, Richard Obexer, Elizabeth L. Bell, Anthony P. Green
Summary: The development and implementation of engineered enzymes and biocatalytic technologies are crucial for achieving sustainable development goals and protecting the environment. By using directed evolution and other methods, engineered enzymes can improve the sustainability of processes such as carbon capture and utilization, bioremediation, plastic deconstruction, and renewable feedstock generation.
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
(2023)
Review
Pharmacology & Pharmacy
Michal Vasina, Jan Velecky, Joan Planas-Iglesias, Sergio M. Marques, Jana Skarupova, Jiri Damborsky, David Bednar, Stanislav Mazurenko, Zbynek Prokop
Summary: Therapeutic enzymes are valuable biopharmaceuticals that have found successful applications in various biomedical fields. The integration of computational approaches and high-throughput experimental technologies has greatly accelerated the identification and engineering of catalytic therapeutic agents, leading to the discovery of new enzymes and the improvement of their properties.
ADVANCED DRUG DELIVERY REVIEWS
(2022)
Article
Chemistry, Multidisciplinary
Andreas Kunzendorf, Guangcai Xu, Mohammad Saifuddin, Thangavelu Saravanan, Gerrit J. Poelarends
Summary: In this study, an enzyme independent of cofactors was engineered for the enantioselective synthesis of cyclopropanes via nucleophilic addition, demonstrating high stereocontrol over both stereocenters. The results highlight the potential of promiscuous enzymes in expanding the biocatalytic repertoire for non-natural reactions.
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
(2021)
Review
Chemistry, Multidisciplinary
Christina Jonsson, Ren Wei, Antonino Biundo, Johan Landberg, Lisa Schwarz Bour, Fabio Pezzotti, Andreea Toca, Les M. Jacques, Uwe T. Bornscheuer, Per-Olof Syren
Summary: Although the separation of individual components from polymer blends in textiles is currently unavailable at a viable scale, biotechnology may provide a solution. However, biotechnological-assisted depolymerization only works for a few synthetic polymer chains, and the use of difficult-to-recycle blended materials is rapidly growing in the textile industry.
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
T. Rehka, Deepti Sharma, Fu Lin, Yeu Khai Choong, Chinchin Lim, Chacko Jobichen, Congqiang Zhang
Summary: The crystal structure of Agrocybe pediades linalool synthase (Ap.LS) was determined, revealing that its specificity is determined by the region of 60-69aa and Tyr299. Mutants of Ap.LS Y299 produced long-chain products. It was also found that microbial terpene synthases with asparagine at a specific position mainly produce cyclized products, while those with a bulky tyrosine produce linear products.
Review
Biochemical Research Methods
Kendreze Holland, John Blazeck
Summary: This review discusses the application of the eukaryotic yeast Saccharomyces cerevisiae as a model host in whole cell biocatalytic conversions, protein evolution, and scientific inquiries into human disease pathogenesis. The review highlights the past and present genome-scale screening platforms and recent advances in biomolecule evolution and high throughput selection and screening strategies. The potential future developments in yeast high throughput engineering are also described.
JOURNAL OF BIOLOGICAL ENGINEERING
(2022)
Review
Biotechnology & Applied Microbiology
Jingjie Jiang, Guangyu Yang, Fuqiang Ma
Summary: Fluorescence-activated droplet sorting (FADS) is a powerful tool for high-throughput screening of enzymes, metabolites, and antibodies. Fluorescence coupling strategies (FCSs) play a key role in developing new FADS methods by linking analyte properties to fluorescence signals. This review highlights recent advances in FCS for different analyte types and discusses successful FADS applications.
BIOTECHNOLOGY ADVANCES
(2023)
Article
Chemistry, Applied
Christoph Schilling, Leesa J. Klau, Finn L. Aachmann, Broder Ruehmann, Jochen Schmid, Volker Sieber
Summary: By using the CRISPR-Cas9 mediated knock-out strategy, a pure fucose-containing polysaccharide named paenan II was successfully isolated from Paenibacillus polymyxa, and its structure was determined using various analytical methods.
CARBOHYDRATE POLYMERS
(2022)
Article
Chemistry, Applied
Tristan Rath, Broder Ruehmann, Jochen Schmid, Volker Sieber
Summary: The study focused on the production of exopolysaccharides (EPS) by Gluconacetobacter sp., with glycerol as a beneficial carbon source compared to glucose, and crude glycerol as a potential substitute. Optimal parameters resulted in a high EPS titer and productivity, with varying peptone concentrations leading to changes in monomer ratios and molecular size distribution. Multiple polysaccharides were identified, with the main one composed of glucose, galactose, mannose, and glucuronic acid, and a minor one of galactose, mannose, and ribose.
CARBOHYDRATE POLYMERS
(2022)
Article
Chemistry, Physical
Ioannis Zachos, Robert Genth, Samuel Sutiono, Matthias Marczynski, Oliver Lieleg, Volker Sieber
Summary: By engineering a glucose dehydrogenase from Saccharolobus solfataricus, researchers were able to adapt it to a thermostable cofactor, carba-NADP(+). Using a microfluidic screening system at elevated temperatures, they discovered variants of the enzyme with significantly increased catalytic speed and efficiency.
Article
Chemistry, Physical
Dhananjai Pangotra, Lenard-Istvan Csepei, Arne Roth, Carlos Ponce de Leon, Volker Sieber, Luciana Vieira
Summary: In this study, a process development was demonstrated to maximize the efficiency of electrochemical production of H2O2, with carbon materials as anodes. The concentration of electrolyte, pH, and the addition of a chemical stabilizer were found to significantly impact the selectivity of water oxidation to H2O2. Additionally, a direct relationship between CO32- ion activity and enhanced H2O2 production was observed, leading to H2O2 concentrations up to 33 mmol L-1 in the anolyte.
APPLIED CATALYSIS B-ENVIRONMENTAL
(2022)
Article
Chemistry, Multidisciplinary
Vivian Pascal Willers, Barbara Beer, Volker Sieber
Summary: Facing increasing mobility, energy demand, and the need to mitigate climate change, developing sustainable and eco-friendly alternatives to fossil fuels is a crucial task for humanity. Biofuels and synthetic fuels show great potential as they can be integrated into existing processes, with synthetic fuels from CO2 and renewable energies being considered the next big step towards a sustainable and ecological future. In our study, we focus on the sustainable production of ethanol and isobutanol, two common biofuels, using methanol and xylose derived from CO2 and lignocellulosic waste streams. Through enzyme and reaction optimization, we successfully produced ethanol or isobutanol from xylose and methanol. Our research opens up the possibility of hybrid production of biofuels and chemicals from CO2 and lignocellulosic waste streams, providing an alternative to conventional routes that rely solely on CO2 or sugars.
Review
Biochemical Research Methods
Mariko Teshima, Vivian Pascal Willers, Volker Sieber
Summary: In the vision of achieving a circular economy that is carbon neutral or even negative, cell-free bioconversion of sustainable and renewable resources has emerged as a promising strategy. In vitro systems have enormous potential, offering technological, ecological, and ethical advantages. Innovative concepts have been developed in cell-free enzymatic conversions to reduce waste production, preserve fossil resources, and recycle industrial pollutants. However, the major challenge in the future will be the transition from concept to industrial application, which requires considerations of productivity, scalability, and cost-effectiveness. In this review, we briefly summarize the latest proof-of-concept cascades focusing on efficient recycling of greenhouse gases using carbon dioxide and other C1 or lignocellulose-derived chemicals, as well as cutting-edge technologies for advancing these concepts to industrial pilot plants.
CURRENT OPINION IN BIOTECHNOLOGY
(2023)
Article
Biochemistry & Molecular Biology
Johanna Radomski, Luciana Vieira, Volker Sieber
Summary: The integration of redox enzymes on electrode surfaces enables the use of renewable energy for highly specific bioelectrochemical synthesis. In this study, the oxidation of glucose to gluconic acid was investigated on a bioanode, combining electrochemical and enzymatic components. The optimized redox film showed a faradaic efficiency of up to 96% for gluconate, and an oxygen-free setup exhibited quantitative electrosynthesis with a gluconate concentration of 6.4±0.25 mmol L-1. Furthermore, the catalase-free anaerobic system showed no production of H2O2 within 24 h, resulting in a highly stable and efficient enzyme performance.
BIOELECTROCHEMISTRY
(2023)
Letter
Chemistry, Multidisciplinary
Dhananjai Pangotra, Arne Roth, Volker Sieber, Luciana Vieira
Summary: The electrochemical production of hydrogen peroxide (H2O2) via 2-electron water oxidation reaction (WOR) has attracted considerable attention. Carbon materials, particularly boron-doped diamond (BDD) electrodes, exhibit high activity for anodic H2O2 generation. However, the high cost of BDD has limited its widespread use. This study explores the use of bipolar plates (BPP) as a low-cost alternative for stable anodic H2O2 generation.
ACS SUSTAINABLE CHEMISTRY & ENGINEERING
(2023)
Article
Chemistry, Multidisciplinary
Samuel Sutiono, Ioannis Zachos, Leandros Paschalidis, Andre Pick, Jakob Burger, Volker Sieber
Summary: In this study, the researchers successfully produced 1,2,4-butanetriol (BTO) from renewable resources, which has versatile applications in pharmaceuticals and plasticizers. By selecting appropriate enzymes and optimizing reaction conditions, they achieved a high titer of 1.2 M (125 g/L) (S)-BTO from D-xylose. Through the addition of an intermediate, they reduced the use of redox cofactors while maintaining a high space-time yield (STY). They also proposed strategies to further increase the titer and yield of BTO.
ACS SUSTAINABLE CHEMISTRY & ENGINEERING
(2023)
Article
Chemistry, Multidisciplinary
Ammar Al-Shameri, Luca Schmermund, Volker Sieber
Summary: O2-dependent biocatalysts are extensively studied and used on the lab scale due to their diversity and functional versatility. However, their reliance on O2 presents technical and economic challenges, such as low O2 transfer rates, low solubility of O2, and limited O2 supply. This mini-review summarizes the latest engineering approaches to address and partially solve these issues.
CURRENT OPINION IN GREEN AND SUSTAINABLE CHEMISTRY
(2023)
Article
Chemistry, Applied
Christoph Schilling, Leesa J. Klau, Finn L. Aachmann, Broder Ruehmann, Jochen Schmid, Volker Sieber
Summary: Combinatorial knock-outs of glycosyltransferases were used to identify the repeating units of two new heteroexopolysaccharides, paenan I and paenan III, produced by P. polymyxa.
CARBOHYDRATE POLYMERS
(2023)
Article
Chemistry, Physical
Vivian Pascal Willers, Manuel Doering, Barbara Beer, Volker Sieber
Summary: Currently, amino acid production is mainly used for livestock feed and food additives. The production relies on sugar fermentation, and it is estimated that the amino acid market will reach ten million tons with a market value over $13 billion USD by 2022. To make amino acid production more sustainable and environmentally friendly, alternative resources and waste streams need to be considered. This study presents a synthetic methanol alanine pathway (MAP) that produces L-alanine from methanol with a maximum theoretical yield of 90% through a cell-free enzymatic cascade. With the increasing possibility of sustainably producing methanol from CO2, this research paves the way for amino acid synthesis based on C1 building blocks and reduces environmental burden.
Review
Biotechnology & Applied Microbiology
Vanessa Wegat, Jonathan T. Fabarius, Volker Sieber
Summary: Global energy-related emissions, especially carbon dioxide, are increasing rapidly. To limit global warming and mitigate climate change, it is crucial to reduce emissions across all sectors. Carbon Capture and Utilization technologies, such as microbial C-1 fermentation, offer innovative ways to close the carbon cycle and establish a circular economy. Synthetic methylotrophic yeasts could play a key role in this process by utilizing alternative feedstock and producing value-added compounds.
BIOTECHNOLOGY FOR BIOFUELS AND BIOPRODUCTS
(2022)
Article
Chemistry, Multidisciplinary
Ioannis Zachos, Samed Guener, Arabella Essert, Peta Lommes, Volker Sieber
Summary: This study focuses on mutational studies on a thermostable glucose dehydrogenase using synthetic cofactors, leading to successful oxidation of different sugars and a significant improvement in system performance.
CHEMICAL COMMUNICATIONS
(2022)
Article
Chemistry, Multidisciplinary
Dhananjai Pangotra, Lenard-Istvan Csepei, Arne Roth, Volker Sieber, Luciana Vieira
Summary: In this study, a process for the continuous flow production of H2O2 at high current densities was developed. The role of CO32- ion activity in enhancing the anodic H2O2 generation was investigated, and optimization of electrolyte flow type and flow rates enabled efficient, stable, and continuous H2O2 production.
