Design of Modular Polyhydroxyalkanoate Scaffolds for Protein Immobilization by Directed Ligation

Wong, Jin Xiang; Rehm, Bernd H. A.

doi:10.1021/acs.biomac.8b01093

Cited by 22 publications

(57 citation statements)

References 88 publications

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“…However, underlying molecular mechanisms of PHA particle formation still remain unknown, which intrinsically limits control of their physicochemical properties. For example, a few studies reported that fusing different proteins to PhaC influences the PHA production yield over biomass, particle size distribution, surface charges, and purity of the target protein (Rubio-Reyes et al, 2016;Gonzalez-Miro et al, 2018a,b;Wong and Rehm, 2018). Decorating PHA particles with proteins using PhaC synthase as an anchoring domain can also cause varying distribution and density of respective proteins on the PHA particles (Hay et al, 2015;Wong and Rehm, 2018).…”

Section: Magnetosome Membrane Protein Mms13mentioning

confidence: 99%

“…For example, a few studies reported that fusing different proteins to PhaC influences the PHA production yield over biomass, particle size distribution, surface charges, and purity of the target protein (Rubio-Reyes et al, 2016;Gonzalez-Miro et al, 2018a,b;Wong and Rehm, 2018). Decorating PHA particles with proteins using PhaC synthase as an anchoring domain can also cause varying distribution and density of respective proteins on the PHA particles (Hay et al, 2015;Wong and Rehm, 2018). Hooks et al (2013) also pointed out that displaying N-acetylneuraminic acid aldolase (NanA) from E. coli on PHA particles through N-and C-terminal fusion of PhaC resulted in varying catalytic performance .…”

Section: Magnetosome Membrane Protein Mms13mentioning

confidence: 99%

“…To circumvent these drawbacks regarding the utilization of the PHA particle technology, our group merged the PhaC fusion technology with the SpyTag/SpyCatcher chemistry (Zakeri et al, 2012), which enable better control over production yields and physicochemical properties (Figure 3A) (Wong and Rehm, 2018). We successfully showed that the SpyTagged proteins could ligate to the SpyCatcher-PhaC coated PHA particles in vitro, and controlled multifunctionality of PHA particles could be achieved using a sequential immobilization strategy.…”

Section: Magnetosome Membrane Protein Mms13mentioning

confidence: 99%

“…Consistency of the particle size and surface charge of functionalized SpyCatcher-coated PHA particles were observed. Function and conformational stability of the ligated proteins were retained or enhanced (Wong and Rehm, 2018). Recently this approach was expanded by developing streamlined processes exploiting the specificity of the SpyTag/SpyCatcher mediated ligation for efficient and cost-effective modular functionalization…”

Section: Magnetosome Membrane Protein Mms13mentioning

confidence: 99%

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Bioengineered Polyhydroxyalkanoates as Immobilized Enzyme Scaffolds for Industrial Applications

Wong

Ogura

Chen

et al. 2020

Front. Bioeng. Biotechnol.

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Enzymes function as biocatalysts and are extensively exploited in industrial applications. Immobilization of enzymes using support materials has been shown to improve enzyme properties, including stability and functionality in extreme conditions and recyclability in biocatalytic processing. This review focuses on the recent advances utilizing the design space of in vivo self-assembled polyhydroxyalkanoate (PHA) particles as biocatalyst immobilization scaffolds. Self-assembly of biologically active enzyme-coated PHA particles is a one-step in vivo production process, which avoids the costly and laborious in vitro chemical cross-linking of purified enzymes to separately produced support materials. The homogeneous orientation of enzymes densely coating PHA particles enhances the accessibility of catalytic sites, improving enzyme function. The PHA particle technology has been developed into a remarkable scaffolding platform for the design of cost-effective designer biocatalysts amenable toward robust industrial bioprocessing. In this review, the PHA particle technology will be compared to other biological supramolecular assembly-based technologies suitable for in vivo enzyme immobilization. Recent progress in the fabrication of biological particulate scaffolds using enzymes of industrial interest will be summarized. Additionally, we outline innovative approaches to overcome limitations of in vivo assembled PHA particles to enable finetuned immobilization of multiple enzymes to enhance performance in multi-step cascade reactions, such as those used in continuous flow bioprocessing.

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Section: Magnetosome Membrane Protein Mms13mentioning

confidence: 99%

Section: Magnetosome Membrane Protein Mms13mentioning

confidence: 99%

Section: Magnetosome Membrane Protein Mms13mentioning

confidence: 99%

Section: Magnetosome Membrane Protein Mms13mentioning

confidence: 99%

See 2 more Smart Citations

Bioengineered Polyhydroxyalkanoates as Immobilized Enzyme Scaffolds for Industrial Applications

Wong

Ogura

Chen

et al. 2020

Front. Bioeng. Biotechnol.

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“…From a biotechnological perspective, these proteins have a high a nity toward the polymeric surface, and they can be usefully employed for immobilization of target proteins by engineering genetic fusions. Indeed, a number of groups have demonstrated successful immobilization of proteins onto PHA supports [11][12][13][14][15] through fusion with PHA-binding proteins, including phasins.…”

Section: Introductionmentioning

confidence: 99%

Bio-specific immobilization of enzymes on electrospun PHB nanofibers

Jang

Park

Lim

et al. 2020

Preprint

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Background Oriented immobilization of enzymes provides substantial advantages in terms of improving the efficiency of enzymatic process as well as enhancing the reusability of enzymes. Phasins have high binding affinity to PHA, and thus can potentially be used as a fusion partner to immobilize enzymes onto PHA supports. However, presently available PHA supports of granular configuration have low surface-area-to-volume ratio, which limits enzyme loading on the supports. In this study, we explored the use of electrospun PHB nanofibers as an alternative support for high density immobilization of a phasin-fused lipase in directed orientations.Results The electrospun PHB nanofibers were successfully used for selective immobilization of the phasin-fused enzyme, and could anchor 120-fold more enzyme than PHB granules of the same weight. The enzyme immobilized onto the PHB nanofibers exhibited exceptionally high stability. Furthermore, via oriented immobilization of the enzymes, PHB nanofibers loaded with the phasin-fused lipase provided 3- to 10-fold higher enzyme activity than the conventional immobilization supports.Conclusions Our approach combines the advantageous features of nanofibrous materials and the regio-specificity of biomolecular interactions for the efficient immobilization of enzymes, which can be widely adopted in the development of various enzymatic processes.

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Alginate Encapsulation of Bioengineered Protein‐Coated Polyhydroxybutyrate Particles: A New Platform for Multifunctional Composite Materials

Ogura

Rehm

2019

Adv Funct Materials

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Immobilization and display of proteins is extensively used to enhance stability and performance of proteins for technical uses such as in biotechnology. Here, self‐assembled nonporous polyhydroxybutyrate (PHB) particles bioengineered to display proteins of interest are subjected to alginate encapsulation processes. The novel composite spheres are fabricated using ionotropic gelation methods. The immunoglobulin G (IgG) binding domain Z and organophosphate hydrolase (OpdA) are attached to PHB particles, and are examples for bioseparation and bioremediation applications, respectively. Alginate microspheres entrapping Z domain coated PHB particles enable flow‐through purification of IgG. Microsphere porosity is pH tunable and at acidic pH IgG is released from Z domains but retained within microspheres. OpdA‐PHB particles are functionally entrapped in alginate microspheres enabling flow‐through substrate conversion. Attachment of functional proteins to PHB particles enhances retention within the alginate microspheres. The hydrophobic PHB particle core within alginate beads provides payload for lipophilic substances, which adsorption kinetics are aligned with a pseudo‐second‐order kinetic model in agreement with the Freundlich isotherm model. This study describes the development of a multifunctional composite material platform based on alginate spheres encapsulating PHB particles that provide payload for lipophilic substances and can be engineered to display protein functions of interest.

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Design of Modular Polyhydroxyalkanoate Scaffolds for Protein Immobilization by Directed Ligation

Cited by 22 publications

References 88 publications

Bioengineered Polyhydroxyalkanoates as Immobilized Enzyme Scaffolds for Industrial Applications

Bioengineered Polyhydroxyalkanoates as Immobilized Enzyme Scaffolds for Industrial Applications

Bio-specific immobilization of enzymes on electrospun PHB nanofibers

Alginate Encapsulation of Bioengineered Protein‐Coated Polyhydroxybutyrate Particles: A New Platform for Multifunctional Composite Materials

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