Affinity-induced covalent protein-protein ligation via the SpyCatcher-SpyTag interaction

Fierer, Jacob O.; Tovar-Herrera, Omar E.; Weinstein, Jonathan; Kahn, Amaranta; Moraïs, Sarah; Mizrahi, Itzhak; Bayer, Edward A.

doi:10.1016/j.greenca.2023.07.001

Cited by 7 publications

(4 citation statements)

References 72 publications

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“…Construction of enzyme complexes is also possible through noncovalent interactions. However, these noncovalent interactions, like cohesin–dockerin, are robust but weak under high temperatures and acidic or alkaline conditions in process environments . To overcome these challenges, we developed a method of covalently locking interactions by integrating the chemistry of three types of Catcher/Tag systems.…”

Section: Resultsmentioning

confidence: 99%

Detoxifying Cyanides Using Cyanase Enzyme Complexes Composed of Carbonic Anhydrase via Irreversible Covalent Bonds

Sun,

Lee,

Han

et al. 2024

J. Agric. Food Chem.

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Cyanase is a possible solution to reduce the environmental impact of cyanide. However, the enzyme’s dependence on HCO3 – limits its industrial applications. To overcome this problem, carbonic anhydrase is utilized in this study. Three types of Catcher/Tag systems were introduced into the cyanase (psCYN) from Pseudomonas stutzeri and the carbonic anhydrase (hmCA) from Hydrogenovibrio marinus to construct enzyme complexes via irreversible covalent bonds. Initially, a cyanase complex with the aid of scaffolding proteins was designed. The results of cyanase complexes using scaffolding proteins were similar to or inferior to those of the two free enzymes. To address this, the two enzymes were manipulated to form a direct bioconjugation without the need for scaffolding proteins. The two enzymes forming a direct conjugation showed activity more than 2.5 times higher than that of cyanase alone. In conclusion, this outcome will contribute to solving problems related to residual cyanides in food and the environment.

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Section: Resultsmentioning

confidence: 99%

Detoxifying Cyanides Using Cyanase Enzyme Complexes Composed of Carbonic Anhydrase via Irreversible Covalent Bonds

Sun,

Lee,

Han

et al. 2024

J. Agric. Food Chem.

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“…The cellulosome is one of the most efficient lignocellulose degradation systems known in nature, with a far higher degradation effi- In addition to free enzymes produced by fungi and bacteria [154], there exists in nature a large multi-enzyme complex produced by anaerobic microorganisms for degrading lignocellulose, known as cellulosome, composed of non-catalytic proteins (scaffoldins) and a variety of glycoside hydrolases [155,156]. The cellulosome is one of the most efficient lignocellulose degradation systems known in nature, with a far higher degradation efficiency than that of free cellulase systems [154,[157][158][159]. The cellulosome assembles various types of cellulases and hemicellulases into a large complex through non-covalent interactions between scaffoldins and enzymes [160,161], creating synergistic and proximity effects among enzymes.…”

Section: Saccharification Processmentioning

confidence: 99%

Composition of Lignocellulose Hydrolysate in Different Biorefinery Strategies: Nutrients and Inhibitors

Wang,

Zhang,

Cui

et al. 2024

Molecules

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The hydrolysis and biotransformation of lignocellulose, i.e., biorefinery, can provide human beings with biofuels, bio-based chemicals, and materials, and is an important technology to solve the fossil energy crisis and promote global sustainable development. Biorefinery involves steps such as pretreatment, saccharification, and fermentation, and researchers have developed a variety of biorefinery strategies to optimize the process and reduce process costs in recent years. Lignocellulosic hydrolysates are platforms that connect the saccharification process and downstream fermentation. The hydrolysate composition is closely related to biomass raw materials, the pretreatment process, and the choice of biorefining strategies, and provides not only nutrients but also possible inhibitors for downstream fermentation. In this review, we summarized the effects of each stage of lignocellulosic biorefinery on nutrients and possible inhibitors, analyzed the huge differences in nutrient retention and inhibitor generation among various biorefinery strategies, and emphasized that all steps in lignocellulose biorefinery need to be considered comprehensively to achieve maximum nutrient retention and optimal control of inhibitors at low cost, to provide a reference for the development of biomass energy and chemicals.

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“…These double‐ and multiple‐dockerin proteins will potentially result in a more complex assembly of cellulosome architectures, but their structure and function have not yet been elucidated. The conformation of these multi‐dockerin regions will not only increase our knowledge of cellulosomes but also provide new components for synthetic biology and biotechnology (Bayer et al, 2008; Feng et al, 2022; Fierer et al, 2023).…”

Section: Introductionmentioning

confidence: 99%

A cellulosomal double‐dockerin module from Clostridium thermocellum shows distinct structural and cohesin‐binding features

Chen,

Yang,

Dong

et al. 2024

Protein Science

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Cellulosomes are intricate cellulose‐degrading multi‐enzymatic complexes produced by anaerobic bacteria, which are valuable for bioenergy development and biotechnology. Cellulosome assembly relies on the selective interaction between cohesin modules in structural scaffolding proteins (scaffoldins) and dockerin modules in enzymes. Although the number of tandem cohesins in the scaffoldins is believed to determine the complexity of the cellulosomes, tandem dockerins also exist, albeit very rare, in some cellulosomal components whose assembly and functional roles are currently unclear. In this study, we characterized the structure and mode of assembly of a tandem bimodular double‐dockerin, which is connected to a putative S8 protease in the cellulosome‐producing bacterium, Clostridium thermocellum. Crystal and NMR structures of the double‐dockerin revealed two typical type I dockerin folds with significant interactions between them. Interaction analysis by isothermal titration calorimetry and NMR titration experiments revealed that the double‐dockerin displays a preference for binding to the cell‐wall anchoring scaffoldin ScaD through the first dockerin with a canonical dual‐binding mode, while the second dockerin module was unable to bind to any of the tested cohesins. Surprisingly, the double‐dockerin showed a much higher affinity to a cohesin from the CipC scaffoldin of Clostridium cellulolyticum than to the resident cohesins from C. thermocellum. These results contribute valuable insights into the structure and assembly of the double‐dockerin module, and provide the basis for further functional studies on multiple‐dockerin modules and cellulosomal proteases, thus highlighting the complexity and diversity of cellulosomal components.

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Affinity-induced covalent protein-protein ligation via the SpyCatcher-SpyTag interaction

Cited by 7 publications

References 72 publications

Detoxifying Cyanides Using Cyanase Enzyme Complexes Composed of Carbonic Anhydrase via Irreversible Covalent Bonds

Detoxifying Cyanides Using Cyanase Enzyme Complexes Composed of Carbonic Anhydrase via Irreversible Covalent Bonds

Composition of Lignocellulose Hydrolysate in Different Biorefinery Strategies: Nutrients and Inhibitors

A cellulosomal double‐dockerin module from Clostridium thermocellum shows distinct structural and cohesin‐binding features

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