Enzyme Immobilization

Vavilala, Sirisha L.; Jain, Ankita; Jain, Amita

doi:10.1016/bs.afnr.2016.07.004

Cited by 218 publications

(54 citation statements)

References 137 publications

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“…Heterogeneous-enzyme hybrids. Heterogeneous-enzyme hybrid catalysts have been applied in the industrial production of chemicals 24,25 . They share many common approaches, benefits and challenges as the heterogeneous-homogeneous ones.…”

Section: Methods and The Pros And Cons Of Constructing Hybridsmentioning

confidence: 99%

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Foundations and strategies of the construction of hybrid catalysts for optimized performances

et al. 2018

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Section: Methods and The Pros And Cons Of Constructing Hybridsmentioning

confidence: 99%

“…General approaches of achieving heterogeneous-enzyme hybrids are support-bound enzymes 27 , entrapment, and support-free crosslinking 24 . Support-bound enzymes consist of enzymes adsorped onto a support or covalently attached to a support (Fig.…”

Section: Methods and The Pros And Cons Of Constructing Hybridsmentioning

confidence: 99%

Foundations and strategies of the construction of hybrid catalysts for optimized performances

et al. 2018

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“…For many biotechnological applications, the activity of proteins displayed on the surface of vegetative B. subtilis should be stable for long periods of time, at least several days if they are to be used as biocatalysts (Homaei, Sariri et al 2013, Sirisha, Jain et al 2016). While a number of display methods have been reported, the durability of displayed heterologous protein activity and how it is affected by extracytoplasmic proteases and solution conditions is not well known.…”

Section: Introductionmentioning

confidence: 99%

Stabilizing displayed proteins on vegetative Bacillus subtilis cells

Huang

Gosschalk

Kim

et al. 2018

Appl Microbiol Biotechnol

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Microbes engineered to display heterologous proteins could be useful biotechnological tools for protein engineering, lignocellulose degradation, biocatalysis, bioremediation and biosensing. Bacillus subtilis is a promising host to display proteins, as this model Gram-positive bacterium is genetically tractable and already used industrially to produce enzymes. To gain insight into the factors that affect displayed protein stability and copy-number, we systematically compared the ability of different protease-deficient B. subtilis strains (WB800, BRB07, BRB08 and BRB14) to display a Cel8A-LysM reporter protein in which the Clostridium thermocellum Cel8A endoglucanase is fused to LysM cell wall binding modules. Whole-cell cellulase measurements and fractionation experiments demonstrate that genetically eliminating extracytoplasmic bacterial proteases improves Cel8A-LysM displays levels. However, upon entering stationary phase, for all protease-deficient strains, the amount of displayed reporter dramatically decreases, presumably as a result of cellular autolysis. This problem can be partially overcome by adding chemical protease inhibitors, which significantly increase protein display levels. We conclude that strain BRB08 is well-suited for stably displaying our reporter protein, as genetic removal of its extracellular and cell wall-associated proteases leads to the highest levels of surface accumulated Cel8A-LysM without causing secretion stress or impairing growth. A two-step procedure is presented that enables the construction of enzyme coated vegetative B. subtilis cells that retain stable cell-associated enzyme activity for nearly 3 days. The results of this work could aid the development of whole cell display systems that have useful biotechnological applications.

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“…The area of deposition is predefined by the selected method of enzyme immobilization. This branch of technology has seen huge success in the past, as it can enhance many properties of enzymes like stability, activity, selectivity, specificity, and resistance to inhibitors [9]. Which of those effects are gained depends not only on the specific enzyme but ofttimes also on sophisticated roadmaps for immobilization.…”

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confidence: 99%

Site-Specific Addressing of Particles and Coatings via Enzyme-Mediated Destabilization

Wedegärtner

Strube

2019

Catalysts

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Enzyme mediated addressing (EMA) is a highly specific and easy-to-apply technology for direction and deposition of particles and coatings on surfaces. Key feature of this process is an enzymatic reaction in direct proximity to the surface, which induces the deposition. The technique has previously shown great success in the handling of biological particles. In this study, addressing of non-biological nanoparticles, in particular plastics and metals, is presented. The respective particles are stabilized by an amphiphilic, enzyme-degradable block copolymer, consisting of poly(ethylene glycol) and poly(caprolactone). After contact with the enzyme pseudomonas lipase, the particles are destabilized, due to the loss of the hydrophilic part of the block copolymer. The lipase is therefore immobilized on glass supports. Immobilization is performed via adsorption or covalent bonding to epoxide groups. All deposition experiments show that addressing of individual particles occurs precisely within the predefined areas of enzyme activity. Depending on the material and reaction conditions, intact nanoparticles or coatings from such can be gained. The quintessence of the study is the indifference of the EMA regarding particle materials. From this rationale, the technique offers near unlimited materials compatibility within a precise, easy-to-apply, and upscalable process.

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Enzyme Immobilization

Cited by 218 publications

References 137 publications

Foundations and strategies of the construction of hybrid catalysts for optimized performances

Foundations and strategies of the construction of hybrid catalysts for optimized performances

Stabilizing displayed proteins on vegetative Bacillus subtilis cells

Site-Specific Addressing of Particles and Coatings via Enzyme-Mediated Destabilization

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