Cellulosomes: Highly Efficient Cellulolytic Complexes

Alves, Victor D.; Fontes, Carlos M. G. A.; Bule, Pedro

doi:10.1007/978-3-030-58971-4_9

Cited by 20 publications

(15 citation statements)

References 167 publications

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“…From the increasing number of described Coh/Doc pairs, we chose the well-characterized type II Coh/Doc pair from Clostridium thermocellum. , The 20 kDa Coh domain was fused to the N-terminus of a tetrameric ADH from Bacillus (Geobacillus) stearothermophilus (∼56 kDa monomer of C-ADH), while the 18 kDa Doc domain was fused to both the N- and C-terminus of a dimeric PLP-dependent ( R )-ωTA from Aspergillus terreus (∼74 kDa monomer of D-ωTA-D) (structures shown in Figure S1). As a proxy element for D-ωTA-D, we also fused Coh domains to both termini of a monomeric enhanced cyan fluorescent protein (∼68 kDa D-ECFP-D).…”

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confidence: 99%

Solid-Phase Assembly of Multienzyme Systems into Artificial Cellulosomes

et al. 2021

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We herein describe a bioinspired solid-phase assembly of a multienzyme system scaffolded on an artificial cellulosome. An alcohol dehydrogenase and an ω-transaminase were fused to cohesin and dockerin domains to drive their sequential and ordered coimmobilization on agarose porous microbeads. The resulting immobilized scaffolded enzymatic cellulosome was characterized through quartz crystal microbalance with dissipation and confocal laser scanning microscopy to demonstrate that both enzymes interact with each other and physically colocalize within the microbeads. Finally, the assembled multifunctional heterogeneous biocatalyst was tested for the one-pot conversion of alcohols into amines. By using the physically colocalized enzymatic system confined into porous microbeads, the yield of the corresponding amine was 1.3 and 10 times higher than the spatially segregated immobilized system and the free enzymes, respectively. This work establishes the basis of a new concept to organize multienzyme systems at the nanoscale within solid and porous immobilization carriers.

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confidence: 99%

Solid-Phase Assembly of Multienzyme Systems into Artificial Cellulosomes

et al. 2021

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“…The scaffoldin itself is also attached to the cell surface via a cohesindockerin domains. The modular nature of the cellulosome inspired the engineering of various designer cellulosomes as well as new multi-enzyme complexes displayed on cell surfaces [91,92]. For in vitro applications, a mini-scaffoldin system was designed for the colocalization of a three-enzyme systems composed of triosephosphate isomerase (TIM), aldolase (ALD), and fructose 1,6-biphosphatase (FBP) which yielded a 33-fold higher catalytic efficiency than the free enzymes [93,94].…”

Section: Co-localization Using Non-covalent Interaction Domains and Tagsmentioning

confidence: 99%

Organizing Multi-Enzyme Systems into Programmable Materials for Biocatalysis

Seo

Schmidt‐Dannert

2021

Catalysts

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Significant advances in enzyme discovery, protein and reaction engineering have transformed biocatalysis into a viable technology for the industrial scale manufacturing of chemicals. Multi-enzyme catalysis has emerged as a new frontier for the synthesis of complex chemicals. However, the in vitro operation of multiple enzymes simultaneously in one vessel poses challenges that require new strategies for increasing the operational performance of enzymatic cascade reactions. Chief among those strategies is enzyme co-immobilization. This review will explore how advances in synthetic biology and protein engineering have led to bioinspired co-localization strategies for the scaffolding and compartmentalization of enzymes. Emphasis will be placed on genetically encoded co-localization mechanisms as platforms for future autonomously self-organizing biocatalytic systems. Such genetically programmable systems could be produced by cell factories or emerging cell-free systems. Challenges and opportunities towards self-assembling, multifunctional biocatalytic materials will be discussed.

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“…Cellulosomes adopt elaborate structures that are built using a series of surface‐displayed scaffoldin proteins that coordinate the binding of the enzyme machinery via dockerin–cohesin domain interactions. They are assembled in modular fashion to create massive polycellulosomal structures that are readily visible by electron microscopy and can harbor >140 distinct dockerin‐borne enzymes in some bacterial species 10,13 . The primary scaffoldin in the C. thermocellum cellulosome, CipA, contains nine type‐I cohesin modules that bind to cellulases harboring type‐I dockerin modules 7 .…”

Section: Introductionmentioning

confidence: 99%

“…Clostridium thermocellum and other anaerobic bacteria within the bacterial orders Clostridiales and Bacteroidales degrade lignocellulose using cellulosomes, multi-enzyme complexes that house an array of enzymes with different substrate specificities (cellulases, hemicellulases, pectinases, esterases etc.). 10 Cellulosome-displaying bacteria degrade lignocellulose more efficiently than microbes that simply secrete cellulases, because enzyme colocalization within the cellulosome promotes enzyme-enzyme synergy, enzyme-proximity enhancement, and cellulose-enzyme-microbe interactions. 11,12 Cellulosomes adopt elaborate structures that are built using a series of surface-displayed scaffoldin proteins that coordinate the binding of the enzyme machinery via dockerin-cohesin domain interactions.…”

Section: Introductionmentioning

confidence: 99%

“…They are assembled in modular fashion to create massive polycellulosomal structures that are readily visible by electron microscopy and can harbor >140 distinct dockerin-borne enzymes in some bacterial species. 10,13 The primary scaffoldin in the C. thermocellum cellulosome, CipA, contains nine type-I cohesin modules that bind to cellulases harboring type-I dockerin modules. 7 The scaffoldin also contains a carbohydrate-binding module (CBM) that tethers the cellulosome complex to its cellulose substrate, as well as a C-terminal type-II dockerin module that anchors the complex to the cell by interacting with surface-displayed proteins that contain type-II cohesin domains.…”

Section: Introductionmentioning

confidence: 99%

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The structure of the Clostridium thermocellum RsgI9 ectodomain provides insight into the mechanism of biomass sensing

et al. 2022

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Clostridium thermocellum is actively being developed as a microbial platform to produce biofuels and chemicals from renewable plant biomass. An attractive feature of this bacterium is its ability to efficiently degrade lignocellulose using surfacedisplayed cellulosomes, large multi-protein complexes that house different types of cellulase enzymes. Clostridium thermocellum tailors the enzyme composition of its cellulosome using nine membrane-embedded anti-σ factors (RsgI1-9), which are thought to sense different types of extracellular carbohydrates and then elicit distinct gene expression programs via cytoplasmic σ factors. Here we show that the RsgI9 anti-σ factor interacts with cellulose via a C-terminal bi-domain unit. A 2.0 Å crystal structure reveals that the unit is constructed from S1C peptidase and NTF2-like protein domains that contain a potential binding site for cellulose. Small-angle X-ray scattering experiments of the intact ectodomain indicate that it adopts a bi-lobed, elongated conformation. In the structure, a conserved RsgI extracellular (CRE) domain is connected to the bi-domain via a proline-rich linker, which is expected to project the carbohydrate-binding unit $160 Å from the cell surface. The CRE and prolinerich elements are conserved in several other C. thermocellum anti-σ factors, suggesting that they will also form extended structures that sense carbohydrates.

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Cellulosomes: Highly Efficient Cellulolytic Complexes

Cited by 20 publications

References 167 publications

Solid-Phase Assembly of Multienzyme Systems into Artificial Cellulosomes

Solid-Phase Assembly of Multienzyme Systems into Artificial Cellulosomes

Organizing Multi-Enzyme Systems into Programmable Materials for Biocatalysis

The structure of the Clostridium thermocellum RsgI9 ectodomain provides insight into the mechanism of biomass sensing

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