Article
Biotechnology & Applied Microbiology
Lydia A. Ogonda, Amelie Saumonneau, Michel Dion, Edward K. Muge, Benson M. Wamalwa, Francis J. Mulaa, Charles Tellier
Summary: This study identified two novel alkaliphilic cellulases, BpGH9 and BpGH48, from Bacillus pumilus. BpGH9, a modular endocellulase, showed high tolerance to alkaline pH and improved activity on crystalline cellulose when engineered as a fusion protein with a CBM3a module.
BIOTECHNOLOGY LETTERS
(2021)
Article
Agricultural Engineering
Wenjun Ying, Yong Xu, Junhua Zhang
Summary: The addition of nonionic surfactants strengthens the interaction between substrate and cellulase, leading to increased adsorption capacity and glucose yield. The gas-liquid interface has a negative impact on enzymatic hydrolysis, with the effect varying depending on enzyme loading, shaking speed, and interfacial size.
INDUSTRIAL CROPS AND PRODUCTS
(2021)
Article
Biochemistry & Molecular Biology
Japheth E. Gado, Brent E. Harrison, Mats Sandgren, Jerry Stahlberg, Gregg T. Beckham, Christina M. Payne
Summary: GH7 family glycoside hydrolases are key enzymes in cellulose degradation, including cellobiohydrolases and endoglucanases. Machine-learning models show that the number of residues in the active-site loops strongly correlates with functional subtype across the GH7 family, with specific residues at certain sequence positions accurately predicting the subtype. Additionally, a random forest model trained on residues in the catalytic domain can predict the presence of a carbohydrate-binding module with high accuracy.
JOURNAL OF BIOLOGICAL CHEMISTRY
(2021)
Article
Biochemistry & Molecular Biology
Nathaniel J. Kuch, Mark E. Kutschke, Alex Parker, Craig A. Bingman, Brian G. Fox
Summary: This study investigates the properties of GH9 enzyme AtCelR from Acetovibrio thermocellus ATCC 27405. Adding an additional carbohydrate binding module (CBM3a) improves its binding affinity and catalytic efficiency for cellulose. However, the specific activity of the engineered enzyme is not increased due to the increase in molecular weight by CBM3a.
JOURNAL OF BIOLOGICAL CHEMISTRY
(2023)
Article
Biotechnology & Applied Microbiology
M. Semenova, A. Gusakov, V. D. Telitsin, A. P. Sinitsyn
Summary: This study investigated the interaction characteristics between different types of LPMOs and cellulase during the enzymatic hydrolysis of cellulose, finding synergism in the initial reaction period but also some antagonism.
APPLIED BIOCHEMISTRY AND MICROBIOLOGY
(2021)
Article
Biotechnology & Applied Microbiology
Veera Hamalainen, Juan De Dios Barajas-Lopez, Yana Berlina, Rafael Alvarez-Rafael, Klara Birikh
Summary: The study successfully addressed the issue of endoglucanase being diverted by other substrates through enzyme engineering and evolution, leading to improved substrate specificity and enhanced activity towards high molecular weight substrates. These findings hold potential applications in the bioeconomy sector, such as second-generation biofuels and biomaterials from lignocellulosic biomass.
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
(2021)
Article
Materials Science, Paper & Wood
Teck-Kim Tang, Yee-Ying Lee, Eng-Tong Phuah, Chin-Ping Tan, Sivaruby Kanagaratnam, Yong Wang, Ling-Zhi Cheong, Ying Li, Nurul Aini Jamalullail, Oi-Ming Lai
Summary: This study investigates the production of nanofibrillated cellulose (NFC) from corn cob using enzymatic pretreatment and high-pressure homogenization process. The incorporation of endoglucanase enzyme softens the corn cob cellulose and prevents instrument blockage. Increasing homogenization cycles and the amount of NFC promote emulsion stability, making it a potential gel-like stabilizer in oil-in-water food emulsion systems.
Article
Biochemistry & Molecular Biology
Parmeshwar Vitthal Gavande, Krishan Kumar, Jebin Ahmed, Arun Goyal
Summary: In this study, the multifunctional endoglucanase RfGH5_4 from Ruminococcus flavefaciens was found to have a (beta/alpha)8-TIM barrel structure through homology modeling. Experimental analyses confirmed the predicted secondary structure of RfGH5_4 and molecular dynamic simulation revealed the stability of its structural fold when binding to a cellulosic ligand. RfGH5_4 showed strong affinity towards cellulosic and hemicellulosic ligands, suggesting its potential in various industrial applications. The study also highlighted the role of specific loops in the catalytic cleft of RfGH5_4 and its monodispersed state and stability.
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
(2023)
Article
Materials Science, Paper & Wood
Martin Nagl, Oskar Haske-Cornelius, Lukas Skopek, Alessandro Pellis, Wolfgang Bauer, Gibson Stephen Nyanhongo, Georg Guebitz
Summary: Paper, with an annual production of over 400 million tons, is the main product of the largest industrially implemented biorefinery process. Enzymes have been used to refine pulp, significantly reducing energy consumption. Different commercial refining enzyme formulations showed varying activities on model substrates, indicating the importance of understanding individual enzyme mechanisms.
Article
Multidisciplinary Sciences
Hongwei Shao
Summary: In this paper, a novel bifunctional cellulase gene cel1 was cloned and expressed in Pichia pastoris GS115. It was found that cel1 has similar sequences to endoglucanase and glucosidase, indicating its multifunctionality. A new gene cel2 was obtained by removing a part of the nucleotide sequence from cel1, which showed 2.1 times higher specific activity. This paper provides important insights into the structure-function relationship of bifunctional enzymes.
Article
Biotechnology & Applied Microbiology
Anica Dadwal, Shilpa Sharma, Tulasi Satyanarayana
Summary: The study successfully cloned and expressed a codon optimized cellobiohydrolase gene from Myceliophthora thermophila in Escherichia coli, demonstrating excellent properties such as stability and tolerance to various environmental conditions. The recombinant MtCel6A is a potential CBH enzyme for industrial applications.
Article
Biochemistry & Molecular Biology
Laure Ayme, Agnes Hebert, Bernard Henrissat, Vincent Lombard, Nathalie Franche, Stephanie Perret, Etienne Jourdier, Senta Heiss-Blanquet
Summary: The study identified six GH9 enzymes from a compost metagenome and characterized three of them, revealing their diverse structures and functional characteristics. These enzymes play a crucial role in bacterial biomass deconstruction.
BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS
(2021)
Article
Polymer Science
Isabel Carrillo-Varela, Claudia Vidal, Sebastian Vidaurre, Carolina Parra, Angela Machuca, Rodrigo Briones, Regis Teixeira Mendonca
Summary: Cold caustic extraction (CCE) with NaOH can be used to remove hemicelluloses and increase cellulose solubility, improving enzymatic saccharification and viscosity reduction of pulps. Eucalyptus pulps are more accessible and easier to hydrolyze by enzymes compared to pine pulps, and the conversion of cellulose I to cellulose II hydrate only increases the saccharification of pine samples.
Article
Chemistry, Applied
Hossam E. Emam, Tharwat Shaheen
Summary: In this study, new stimuli-responsive hybrid hydrogels were successfully created by incorporating succinylated cellulose nanocrystals (Su-CNC) into the hydrogel network. The hydrogels exhibited pH and thermo-responsive characteristics, with significant swelling behavior at pH 6 and shrinkage at temperatures above 35 degrees C.
CARBOHYDRATE POLYMERS
(2022)
Article
Biotechnology & Applied Microbiology
Hai-Yan Zhou, Xiao-Nan Yi, Qi Chen, Jian-Bao Zhou, Shu-Fang Li, Xue Cai, De-Shui Chen, Xin-Ping Cheng, Mian Li, Hong-Yan Wang, Kai-Qian Chen, Zhi-Qiang Liu, Yu-Guo Zheng
Summary: In this study, a novel endoglucanase (CgEndo) from Colletotrichum graminicola was modified by site-directed mutagenesis to improve its catalytic efficiency. Two mutants, Y63S and N20D/S113T, showed significantly increased enzyme activities. The mutations improved the proximity between the enzyme and its substrate, leading to higher enzyme activities and catalysis efficiency. These findings lay an important foundation for further engineering of the endoglucanase and its practical application in efficient degradation of cellulosic biomass.
ENZYME AND MICROBIAL TECHNOLOGY
(2022)
Article
Biochemistry & Molecular Biology
Malene Billeskov Keller, Silke Flindt Badino, Nanna Rojel, Trine Holst Sorensen, Jeppe Kari, Brett McBrayer, Kim Borch, Benedikt M. Blossom, Peter Westh
Summary: The study found that C1-oxidizing LPMOs have varying effects on the activity of different reducing-end cellobiohydrolases, with oxidation reducing enzyme activity and substrate complexation rate. This finding suggests that C1-oxidized chain ends are not ideal sites for attack by reducing-end cellobiohydrolases, which could be relevant for optimizing industrial enzyme cocktails.
JOURNAL OF BIOLOGICAL CHEMISTRY
(2021)
Article
Multidisciplinary Sciences
Jeppe Kari, Gustavo A. Molina, Kay S. Schaller, Corinna Schiano-di-Cola, Stefan J. Christensen, Silke F. Badino, Trine H. Sorensen, Nanna S. Rojel, Malene B. Keller, Nanna Rolsted Sorensen, Bartlomiej Kolaczkowski, Johan P. Olsen, Kristian B. R. M. Krogh, Kenneth Jensen, Ana M. Cavaleiro, Gunther H. J. Peters, Nikolaj Spodsberg, Kim Borch, Peter Westh
Summary: Enzyme reactions at interfaces are common in both Nature and industrial applications, but there is a lack of a general kinetic framework for interfacial enzymes. This study characterizes 83 cellulases and identifies a linear free energy relationship between ligand binding strength and maximal turnover, which may provide insights into cellulolytic mechanisms and aid in the selection of technical enzymes.
NATURE COMMUNICATIONS
(2021)
Article
Chemistry, Physical
Kay S. Schaller, Gustavo Avelar Molina, Jeppe Kari, Corinna Schiano-di-Cola, Trine Holst Sorensen, Kim Borch, Gunther H. J. Peters, Peter Westh
Summary: Deposition of enzyme sequences exceeds the possibilities of experimental characterization. This study presents a virtual screening approach that predicts kinetic parameters of enzymes based on their sequences. Using fungal cellulases as an example, the predicted kinetic parameters showed good agreement with experimental results, and one enzyme displayed exceptional performance.
Article
Biochemistry & Molecular Biology
Sune Schubert, Kay Schaller, Jenny Arnling Baath, Cameron Hunt, Kim Borch, Kenneth Jensen, Jesper Brask, Peter Westh
Summary: Bioprocessing of polyester waste is a promising approach for achieving a cyclic plastic economy. This study focuses on the enzymatic breakdown of poly(ethylene terephthalate) (PET) and reveals the crucial role of endo-lytic activity in overall enzyme performance. The release of soluble PET fragments with two or three aromatic rings through endo-lytic activity leads to efficient breakdown of the polymer in aqueous bulk, contributing to the formation of monoaromatic products.
Review
Biochemical Research Methods
Jeppe Kari, Kay Schaller, Gustavo A. Molina, Kim Borch, Peter Westh
Summary: The recent breakthrough in all-atom, protein structure prediction has opened up new computational approaches in enzyme design. These approaches have the potential to significantly contribute to the development of technical biocatalysts and facilitate the transition to more sustainable industries. This article discusses the implementation of linear free energy relationships (LFERs) and the Sabatier Principle in algorithms for predicting enzyme kinetic parameters and performance based on model structures.
CURRENT OPINION IN BIOTECHNOLOGY
(2022)
Review
Biochemistry & Molecular Biology
Morten Sorlie, Malene Billeskov Keller, Peter Westh
Summary: In nature, the enzymatic degradation of recalcitrant polysaccharides involves the collaborative action of glycoside hydrolases (GHs) and lytic polysaccharide monooxygenases (LPMOs) through different mechanisms. GHs use a hydrolytic approach, while LPMOs are oxidative. The interplay between LPMOs and GHs can provide rate enhancements and improved degradation of polysaccharides, although the exact magnitude of the enhancements depends on the nature of the specific GH and LPMO. The understanding of this interplay is important for enhancing enzymatic polysaccharide degradation.
ESSAYS IN BIOCHEMISTRY
(2023)
Review
Biochemistry & Molecular Biology
Yu Tian, Yu Wang, Yuyue Zhong, Marie Sofie Moller, Peter Westh, Birte Svensson, Andreas Blennow
Summary: Enzymatic hydrolysis of starch granules is fundamental for understanding starch degradation in plant cells, utilizing starch as an energy resource in foods, and developing efficient saccharification processes. However, the kinetics of hydrolysis for solid substrates like starch granules are slower and more complex. The multifaceted nature of the substrate surface and its changing properties during hydrolysis pose challenges for analysis.
Article
Biochemical Research Methods
Bartlomiej M. Kolaczkowski, Olga V. Moroz, Elena Blagova, Gideon J. Davies, Marie Sofie Moller, Anne S. Meyer, Peter Westh, Kenneth Jensen, Keith S. Wilson, Kristian B. R. M. Krogh
Summary: Many eukaryotic proteins are N-glycosylated with high-mannose N-glycans and alpha-1,6-mannan backbones. GH92 alpha-mannosidases release mannose residues from these N-glycans, allowing alpha-endomannanases to degrade the alpha-mannan backbone. The structure and function of a multidomain GH92 alpha-mannosidase CBM have not been characterized. This study provides insights into the structure and function of a full-length five-domain GH92 alpha-mannosidase from Neobacillus novalis, demonstrating the importance of CBMs for the overall structural integrity of the enzyme.
ACTA CRYSTALLOGRAPHICA SECTION D-STRUCTURAL BIOLOGY
(2023)
Article
Chemistry, Multidisciplinary
Jenny Arnling Baath, Kenneth Jensen, Kim Borch, Peter Westh, Jeppe Kari
Summary: Investigated the kinetics of two PET hydrolases at different concentrations of CTAB and found that CTAB consistently lowered the strength of enzyme-PET interactions. The effect of CTAB on enzymatic turnover was biphasic, with initial promotion and subsequent suppression. The correlation with maximal turnover at an intermediate binding strength was in accordance with the Sabatier principle. Suggested that future discovery efforts should consider enzymes with weakened substrate binding due to their limiting effect on catalytic performance.
Article
Biochemistry & Molecular Biology
Bartlomiej M. Kolaczkowski, Christian Jorgensen, Nikolaj Spodsberg, Mary A. Stringer, Nitin T. Supekar, Parastoo Azadi, Peter Westh, Kristian B. R. M. Krogh, Kenneth Jensen
Summary: Glycoengineering allows control over glycosylation patterns to generate new glycoprotein variants with desired properties. Challenges include glycan heterogeneity and nonnative glycan chains introduced by heterologous protein expression. Partial trimming or complete removal of glycan chains can address these challenges.
Article
Chemistry, Physical
Hiroyo Ohgi, Hiroshi Imamura, Tomonari Sumi, Keiko Nishikawa, Yoshikata Koga, Peter Westh, Takeshi Morita
Summary: The study found that in the TFE-H2O mixture, concentration fluctuations of TFE molecules and various interactions between molecules have a significant impact on helix formation of proteins/peptides.
PHYSICAL CHEMISTRY CHEMICAL PHYSICS
(2021)
