A novel method for simultaneous purification and immobilization of a xylanase-lichenase chimera via SpyTag/SpyCatcher spontaneous reaction

Lin, Yuxia; Jin, Wenhui; Wang, Jindan; Cai, Zhengwen; Wu, Shu-Yu; Zhang, Guangya

doi:10.1016/j.enzmictec.2018.04.007

Cited by 28 publications

(13 citation statements)

References 52 publications

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“…Anchoring onto a single layer of graphene using SpyTag has been developed for enhancing cryoelectron microscopy structure determination at atomic resolution. 13 SpyTag-linked proteins may be anchored to silica particles assembled through biomimetic silicication 14 or to plastic particles (polyhydroxyalkanoate, PHA) synthesised inside the cell 15 ( Fig. 2A).…”

Section: Application Areas 21 Anchoring To Surfaces or Particlesmentioning

confidence: 99%

Power to the protein: enhancing and combining activities using the Spy toolbox

2020

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Section: Application Areas 21 Anchoring To Surfaces or Particlesmentioning

confidence: 99%

Power to the protein: enhancing and combining activities using the Spy toolbox

2020

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“…The construct was further explored for the detection of ovalbumin in ELISA by conjugation of SpyTagged Nanoluc and protein G on the scaffold [ 45 ]. Another research group has developed a single-step method for the purification and immobilization of xylanase-lichenase chimera [ 46 ]. SpyTag was fused between xylanase and lichenase to form a chimera that covalently binds to SpyCatcher-elastin like polypeptides (purification tag) via in vitro spontaneous SpyTag-SpyCatcher reaction and simultaneously self-assembled to form insoluble active particles during purification process which serves as immobilized enzyme complex.…”

Section: Strategies Of Enzyme Immobilization On a Protein Scaffoldmentioning

confidence: 99%

“…This immobilized enzyme chimera improved the stability by retaining 44% and 56% activities of xylanase and lichenase respectively even after 10 subsequent reaction cycles. However, a 1.7-fold (xylanase) and 1.1-fold (lichenase) decrease in the catalytic efficiency of immobilized enzyme was found [ 46 ].…”

Section: Strategies Of Enzyme Immobilization On a Protein Scaffoldmentioning

confidence: 99%

Protein scaffolds: A tool for multi-enzyme assembly

Gad

Ayakar

2021

Biotechnology Reports

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Highlights Protein scaffold acts as a backbone with several interacting domains and ligands for the integration of specific enzymes. Different protein-enzyme ligation strategies are used for the co-localization of enzymes. Scaffolds permit control over spatial organization, enzyme stoichiometry, and proximity. Improvement in pathway flux and product yield can be achieved with scaffold-based multi-enzyme complex. Various analytical tools are utilized to study the multi-enzyme assembly process.

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“…The reaction could occur rapidly under mild conditions with high efficiency and specificity, and the covalent bond between them is robust to diverse harsh conditions [14][15][16][17]. Thus, SpyTag/SpyCatcher is deemed a novel and efficient molecular adhesion that plays a vital role in the immobilization of enzymes [18][19][20].…”

Section: Introductionmentioning

confidence: 99%

A novel all-in-one strategy for purification and immobilization of β-1,3-xylanase directly from cell lysate as active and recyclable nanobiocatalyst

et al. 2021

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Background Exploring a simple and versatile technique for direct immobilization of target enzymes from cell lysate without prior purification is urgently needed. Thus, a novel all-in-one strategy for purification and immobilization of β-1,3-xylanase was proposed, the target enzymes were covalently immobilized on silica nanoparticles via elastin-like polypeptides (ELPs)-based biomimetic silicification and SpyTag/SpyCatcher spontaneous reaction. Thus, the functional carriers that did not require the time-consuming surface modification step were quickly and efficiently prepared. These carriers could specifically immobilize the SpyTag-fused target enzymes from the cell lysate without pre-purification. Results The ELPs-SpyCatcher hardly leaked from the carriers (0.5%), and the immobilization yield of enzyme was up to 96%. Immobilized enzyme retained 85.6% of the initial activity and showed 88.6% of the activity recovery. Compared with free ones, the immobilized β-1,3-xylanase showed improved thermal stability, elevated storage stability and good pH tolerance. It also retained more than 70.6% of initial activity after 12 reaction cycles, demonstrating its excellent reusability. Conclusions The results clearly highlighted the effectiveness of the novel enzyme immobilization method proposed here due to the improvement of overall performance of immobilized enzyme in respect to free form for the hydrolysis of macromolecular substrates. Thus, it may have great potential in the conversion of algae biomass as well as other related fields.

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A novel method for simultaneous purification and immobilization of a xylanase-lichenase chimera via SpyTag/SpyCatcher spontaneous reaction

Cited by 28 publications

References 52 publications

Power to the protein: enhancing and combining activities using the Spy toolbox

Power to the protein: enhancing and combining activities using the Spy toolbox

Protein scaffolds: A tool for multi-enzyme assembly

A novel all-in-one strategy for purification and immobilization of β-1,3-xylanase directly from cell lysate as active and recyclable nanobiocatalyst

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