Review
Biotechnology & Applied Microbiology
Zuzana Meszaros, Pavlina Nekvasilova, Pavla Bojarova, Vladimir Kren, Kristyna Slamova
Summary: With the establishment of glycosynthase and transglycosidase technology by genetic engineering, glycosidases have become competitive synthetic tools in industrial biotechnology, offering advantages such as easy production, affordability, robustness, and substrate versatility.
BIOTECHNOLOGY ADVANCES
(2021)
Review
Biotechnology & Applied Microbiology
Zuzana Meszaros, Pavlina Nekvasilova, Pavla Bojarova, Vladimir Kren, Kristyna Slamova
Summary: Traditionally, glycosidases have been underutilized in industry, primarily used for cleaving unwanted carbohydrates, but with advancements in genetic engineering, they are now competitive synthetic tools capable of producing various chemicals, such as chitooligomers and glycosylated flavonoids. Exciting developments are on the horizon, including the production of human milk oligosaccharides and remodeled antibodies.
BIOTECHNOLOGY ADVANCES
(2021)
Article
Biochemistry & Molecular Biology
Yihao Liu, Junwen Ma, Ran Shi, Ting Li, Qiaojuan Yan, Zhengqiang Jiang, Shaoqing Yang
Summary: A beta-N-acetylhexosaminidase gene (CmHex187) from Catenibacterium mitsuokai was successfully expressed in Escherichia coli, showing strong transglycosylation activity with a high conversion ratio in the synthesis of an important HMO component, lacto-N-triose II (LNT2). The enzyme displayed optimal pH and temperature, promoting the growth of tested Lactobacillus and Bifidobacteria strains, making it a promising candidate for LNT2 production.
PROCESS BIOCHEMISTRY
(2021)
Article
Biochemistry & Molecular Biology
Patrick Weber, Zuzana Meszaros, Pavla Bojarova, Manuel Ebner, Roland Fischer, Vladimir Kren, Natalia Kulik, Philipp Mueller, Miluse Vlachova, Kristyna Slamova, Arnold E. Stuetz, Martin Thonhofer, Ana Torvisco, Tanja M. Wrodnigg, Andreas Wolfsgruber
Summary: A new class of compounds that inhibit the de-O-glycosylation of proteins has been discovered. These compounds, highly substituted diaminocyclopentanes, are selective and reversible inhibitors of O-beta-N-acetyl-D-glucosaminidase (O-GlcNAcase), and show impressive biological activities. They could serve as promising leads for the development of anti-tau-phosphorylation agents to potentially ameliorate Alzheimer's disease.
BIOORGANIC CHEMISTRY
(2023)
Article
Food Science & Technology
Masahiro Kurakake, Yukari Amai
Summary: In this study, human milk oligosaccharides with various health functions were synthesized through transglycosylation reaction of beta-N-acetylhexosaminidase. New transfer products were developed through this reaction. Microorganisms with good transglycosylation activity were isolated from soil. It was found that Metarhizium sp. A34 beta-N-acetylhexosaminidase could produce various oligosaccharides under substrate degradation conditions.
JOURNAL OF FOOD SCIENCE
(2022)
Article
Microbiology
Yan Chen, Ning Zhou, Xueman Chen, Guoguang Wei, Alei Zhang, Kequan Chen, Pingkai Ouyang
Summary: In this study, a gene encoding beta-N-acetylglucosaminidase, designated NAGaseA, was cloned and functionally expressed in Escherichia coli. NAGaseA showed both transglycosylation and hydrolytic activities, making it a potential candidate for GlcNAc production.
FRONTIERS IN MICROBIOLOGY
(2022)
Article
Biochemistry & Molecular Biology
Lucas Matheus Soares Pereira, Aline Vianna Bernardi, Luis Eduardo Gerolamo, Wellington Ramos Pedersoli, Claudia Batista Carraro, Roberto do Nascimento Silva, Sergio Akira Uyemura, Taisa Magnani Dinamarco
Summary: Concern over environmental impacts has led to efforts to replace fossil fuels with biofuels, such as ethanol. However, additional investments in technologies like second generation ethanol are necessary to meet the growing demand. Currently, the production is not economically feasible due to high costs of enzymes used in the saccharification stage of biomass.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2023)
Article
Biotechnology & Applied Microbiology
Yihao Liu, Qiaojuan Yan, Junwen Ma, Jianyu Wang, Zhengqiang Jiang, Shaoqing Yang
Summary: In this study, a mutant enzyme (mHaHex74) with improved transglycosylation activity was obtained by directed evolution and site-saturation mutagenesis of HaHex74. mHaHex74 showed higher conversion ratio and activity at different pH and temperature conditions, indicating its potential for the enzymatic synthesis of human milk oligosaccharides.
ENZYME AND MICROBIAL TECHNOLOGY
(2023)
Article
Genetics & Heredity
Pan Wang, Congcong Tang, Yannan Liu, Jing Yang, Daidi Fan
Summary: This study successfully expressed a thermostable β-glycosidase from Sulfolobus solfataricus in Pichia pastoris and secreted it into fermentation broth. The optimized enzyme showed the highest catalytic efficiency at 80°C and pH 6.0, and its activity was significantly enhanced in the presence of Li+. The recombinant enzyme could completely convert the ginsenoside substrate to the rare ginsenoside Compound K and exhibited extraordinary tolerance against high substrate concentrations.
Article
Biochemistry & Molecular Biology
Xing Qin, YanZhe Xin, Xiaoyun Su, Xiaolu Wang, Jie Zhang, Tao Tu, Yaru Wang, Bin Yao, Huoqing Huang, Huiying Luo
Summary: The study identified two thermostable chitinolytic enzymes from Caldicellulosiruptor acetigenus, CaChiT and CaHex, which showed optimal activities at high temperatures and broad pH stability. These enzymes were active on various chitin substrates and exhibited a synergistic effect in chitin degradation, indicating their potential for industrial applications in GlcNAc production.
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
(2021)
Article
Chemistry, Physical
Michael Kotik, Katerina Brodsky, Petr Halada, Hana Javurkova, Helena Pelantova, Dorota Konvalinkova, Pavla Bojarova, Vladimir Kren
Summary: Two newly identified wild-type rutinosidases from glycoside hydrolase family 5-23 can glycosylate inorganic azide with rutin as a donor, producing rutinosyl alpha-azide. The catalytic nucleophile mutant of rutinosidase from Aspergillus niger, also belonging to GH5-23, was able to synthesize rutinosyl beta-azide. Thus, both anomers of rutinosyl azide can be synthesized at a preparatory scale using either wild-type or mutant rutinosidases of GH5-23.
CATALYSIS COMMUNICATIONS
(2021)
Article
Agriculture, Multidisciplinary
Ju-Hee Cha, Minsun Hong, Chang-Jun Cha
Summary: In this study, a fungal beta-glycosidase belonging to subfamily 4 of GH family 30 (GH30) was characterized with hydrolysis and transglycosylation activities towards various substrates, representing the first fungal beta-glycosidase with transglycosylation activities in subfamily 4 of GH30.
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
(2021)
Article
Biochemistry & Molecular Biology
Yi Zhu, Gerald W. Hart
Summary: O-GlcNAc is a dynamic post-translational modification that affects protein functions. In this study, a dual RNA-aptamer-based approach was developed to selectively increase O-GlcNAcylation of (3-catenin without affecting other OGT substrates. The increased O-GlcNAcylation of (3-catenin stabilized the protein, altered its interactions with (3-TrCP and EZH2, and resulted in significant changes in the transcriptome. The use of riboswitches or an inducible expression system coupled with aptamers allowed for inducible regulation of protein-specific O-GlcNAcylation.
Article
Chemistry, Applied
Jing Li, Kunpeng Gao, Francesco Secundo, Xiangzhao Mao
Summary: Chitin is converted to GlcNAc by chitinases, essential for food and pharmaceutical industries. Two novel beta-N-acetylglucosaminidases were cloned and characterized, enabling the one-pot production of pure GlcNAc from chitin through enzymatic reactions.
Article
Biochemistry & Molecular Biology
Yun Ge, Daniel H. Ramirez, Bo Yang, Alexandria K. D'Souza, Chanat Aonbangkhen, Stephanie Wong, Christina M. Woo
Summary: In this study, a nanobody-fused split O-GlcNAcase (OGA) was developed as an O-GlcNAc eraser for selectively deglycosylating target proteins in cells. Through systematic cellular optimization, a split OGA with reduced inherent deglycosidase activity was identified for selective removal of O-GlcNAc from target proteins guided by nanobody. The system demonstrated generality by using four nanobodies against five target proteins, allowing the study of O-GlcNAc impact on transcription factors c-Jun and c-Fos.
NATURE CHEMICAL BIOLOGY
(2021)
Review
Biotechnology & Applied Microbiology
Viktoria Heine, Carina Dey, Pavla Bojarova, Vladimir Kren, Lothar Elling
Summary: Galectins are a family of lectins that modulate cell events by cross-linking the glycan structures of cell membranes and extracellular matrix components. Targeting galectins with glycomaterials for diagnostics or therapy is a focus of research, and this review explores different methods to identify potential glycomaterials for translating galectin targeting from molecular level to in vivo assays.
BIOTECHNOLOGY ADVANCES
(2022)
Article
Biochemistry & Molecular Biology
Mariann Kiss, Istvan Timari, Terez Barna, Zuzana Meszaros, Kristyna Slamova, Pavla Bojarova, Vladimir Kren, Joseph M. Hayes, Laszlo Somsak
Summary: The inhibition of human O-linked beta-N-acetylglucosaminidase (hOGA) and human lysosomal hexosaminidases (hHexA and hHexB) is significant in the treatment of various diseases. A series of compounds were synthesized and evaluated against these enzymes, revealing a potent inhibitor for hOGA.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2022)
Article
Chemistry, Medicinal
David Vrbata, Marcela Filipova, Marina R. Tavares, Jakub Cerveny, Miluse Vlachova, Milada Sirova, Helena Pelantova, Lucie Petraskova, Ladislav Bumba, Rafal Konefal, Tomas Etrych, Vladimir Kren, Petr Chytil, Pavla Bojarova
Summary: Galectin-3 plays a role in cancer-related metabolic processes. Inhibiting overexpressed Galectin-3, such as with beta-galactoside-based inhibitors, holds promise for cancer treatment. A library of inhibitors was synthesized and displayed on a suitable biocompatible carrier to enhance affinity for Gal-3. One of the candidates exhibited strong inhibitory effects in cancer cells and shows potential for Gal-3-associated cancer therapy.
JOURNAL OF MEDICINAL CHEMISTRY
(2022)
Article
Chemistry, Applied
Zuzana Meszaros, Lucie Petraskova, Natalia Kulik, Helena Pelantova, Pavla Bojarova, Vladimir Kren, Kristyna Slamova
Summary: Fungal beta-N-acetylhexosaminidases are important tools for the synthesis of natural and modified oligosaccharides and glycoconjugates. In this study, the enzyme from Aspergillus oryzae was engineered to shift its reaction from hydrolysis to transglycosylation. Nine mutant variants of the enzyme were designed and tested, and one variant (V306W/Y445N AoHex) showed superior transglycosidase activity with a transglycosylation-to-hydrolysis ratio greater than 110, which is unique among similar mutants.
ADVANCED SYNTHESIS & CATALYSIS
(2022)
Article
Biochemistry & Molecular Biology
Valeria Pingitore, Macarena Martinez-Bailen, Ana T. Carmona, Zuzana Meszaros, Natalia Kulik, Kristyna Slamova, Vladimir Kren, Pavla Bojarova, Inmaculada Robina, Antonio J. Moreno-Vargas
Summary: Two libraries of mono- and dimeric pyrrolidine iminosugars were synthesized and screened for inhibition of enzymes from Jack beans and human lysosomal hexosaminidases, resulting in the identification of potent and selective inhibitors for human nucleocytoplasmic beta-N-acetylglucosaminidase.
BIOORGANIC CHEMISTRY
(2022)
Article
Biochemistry & Molecular Biology
Pavlina Nekvasilova, Michaela Hovorkova, Zuzana Meszaros, Lucie Petraskova, Helena Pelantova, Vladimir Kren, Kristyna Slamova, Pavla Bojarova
Summary: Enzymatic synthesis is a biocompatible method for the synthesis of complex compounds, including human milk oligosaccharides. Previous studies have been limited to analytical reactions, but this study demonstrates the successful synthesis of three tetrasaccharides derived from lacto-N-tetraose using engineered glycosidases. The reaction products were isolated and characterized.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2022)
Article
Chemistry, Multidisciplinary
Katerina Brodsky, Kristyna Kanova, Dorota Konvalinkova, Kristyna Slamova, Helena Pelantova, Katerina Valentova, Pavla Bojarova, Vladimir Kren, Lucie Petraskova
Summary: Enzymatic sulfation is an important and understudied topic, and finding suitable sulfate donors is a major challenge. This study investigated the impact of different sulfate donors on enzyme-catalyzed sulfation, and evaluated the sulfated compounds obtained.
Article
Biochemistry & Molecular Biology
Michaela Hovorkova, Jakub Cerveny, Ladislav Bumba, Helena Pelantova, Josef Cvacka, Vladimir Kren, Olivier Renaudet, David Goyard, Pavla Bojarova
Summary: A new type of glycoconjugate with low nanomolar affinity was developed to inhibit galectins of interest for biomedical research. The presentation of carbohydrate ligands on specific structural scaffolds was found to impact the affinity and selectivity to different galectins.
BIOORGANIC CHEMISTRY
(2023)
Article
Chemistry, Organic
Dorota Konvalinkova, Frantisek Dolnicek, Michaela Hovorkova, Jakub Cerveny, Ondrej Kundrat, Helena Pelantova, Lucie Petraskova, Josef Cvacka, Margarita Faizulina, Beena Varghese, Petr Kovaricek, Vladimir Kren, Pavel Lhotak, Pavla Bojarova
Summary: This study reports the synthesis of potential galectin ligands based on the calix[4]arene core and investigates the influence of linkers and core structure on their affinity. Competitive assays using ELISA and biolayer interferometry demonstrate significant differences in affinity based on linker length and the position of the triazole unit. Atomic force microscopy detects the formation of supramolecular clusters.
ORGANIC & BIOMOLECULAR CHEMISTRY
(2023)
Article
Biochemistry & Molecular Biology
Patrick Weber, Zuzana Meszaros, Pavla Bojarova, Manuel Ebner, Roland Fischer, Vladimir Kren, Natalia Kulik, Philipp Mueller, Miluse Vlachova, Kristyna Slamova, Arnold E. Stuetz, Martin Thonhofer, Ana Torvisco, Tanja M. Wrodnigg, Andreas Wolfsgruber
Summary: A new class of compounds that inhibit the de-O-glycosylation of proteins has been discovered. These compounds, highly substituted diaminocyclopentanes, are selective and reversible inhibitors of O-beta-N-acetyl-D-glucosaminidase (O-GlcNAcase), and show impressive biological activities. They could serve as promising leads for the development of anti-tau-phosphorylation agents to potentially ameliorate Alzheimer's disease.
BIOORGANIC CHEMISTRY
(2023)
Article
Chemistry, Multidisciplinary
Patrick Weber, Zuzana Meszaros, Denis Jagecic, Valentina Hribljan, Dinko Mitrecic, Pavla Bojarova, Kristyna Slamova, Jiri Vrba, Natalia Kulik, Vladimir Kren, Arnold E. Stuetz
Summary: Potent and selective aminocyclopentane-derived inhibitors of human O-N-acetyl-beta-d-glucosaminidase (OGA) have been developed for the treatment of Alzheimer's disease. Compound 13, for example, showed nanomolar OGA inhibition and 92,000-fold selectivity over human HexB. It was non-toxic and increased protein O-GlcNAcylation in murine neural cell culture, providing new alternatives for tauopathies treatment.
CHEMICAL COMMUNICATIONS
(2022)
Review
Biotechnology & Applied Microbiology
Zhi-Min Zhao, Zhi-Hua Liu, Tongtong Zhang, Rongqian Meng, Zhiqun Gong, Yibing Li, Jing Hu, Arthur J. Ragauskas, Bing-Zhi Li, Ying-Jin Yuan
Summary: Bioconversion of bioresources/wastes using Rhodococcus is a promising approach for producing lipid-derived bioproducts. However, bioconversion involves living cells and is a complex system that requires further understanding. This review highlights the potential of Rhodococcus for producing lipids from various wastes and presents genetic modifications using emerging tools for promoting bioconversion. Additionally, strategies for enhancing the bioaccessibility of lignin and its derived aromatics and synthesizing biodiesel and aviation fuels from lignin and lipids are discussed. The synergistic modification of microbial strains, substrates, and fermentation processes is proposed for sustainable biological lignin valorization.
BIOTECHNOLOGY ADVANCES
(2024)
Review
Biotechnology & Applied Microbiology
Maddy L. Newby, Joel D. Allen, Max Crispin
Summary: A key aspect of successful viral vaccine design is the elicitation of neutralizing antibodies targeting viral attachment and fusion glycoproteins. Glycosylation can influence the antigenicity and immunogenicity of vaccines, with native-like glycosylation potentially enhancing immunogenicity. Understanding viral glycosylation can guide the development of future vaccines in various enveloped viruses.
BIOTECHNOLOGY ADVANCES
(2024)
Review
Biotechnology & Applied Microbiology
Uganeeswary Suparmaniam, Man Kee Lam, Jun Wei Lim, Inn Shi Tan, Bridgid Lai Fui Chin, Siew Hoong Shuit, Steven Lim, Yean Ling Pang, Peck Loo Kiew
Summary: Microalgae have the ability to synthesize high-value phytochemicals, and subjecting them to abiotic stress can enhance the accumulation of valuable metabolites within their cells. This study explores the effects of manipulating abiotic stress on microalgae yields and discusses approaches to increase HVPCs production using abiotic stress.
BIOTECHNOLOGY ADVANCES
(2024)
Review
Biotechnology & Applied Microbiology
Pei Zhou, Cong Gao, Wei Song, Wanqing Wei, Jing Wu, Liming Liu, Xiulai Chen
Summary: The engineering status of protein is crucial for the efficient bioproduction in microbial cell factories. Strategies such as protein engineering, protein modification, and protein assembly can effectively modify the protein status and enhance production efficiency.
BIOTECHNOLOGY ADVANCES
(2024)
