3D Printed Polylactic Acid Well-Plate for Multi-enzyme Immobilization

Gkantzou, Elena; Skonta, Anastasia; Vasios, Andreas-Georgios; Stamatis, Haralambos

doi:10.1007/978-1-0716-2269-8_10

Cited by 3 publications

(5 citation statements)

References 10 publications

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“…PLA scaffolds were designed according to our previous reports using the Fusion 360 CAD program from Autodesk (USA) [29,30]. Briefly, well plates were designed with a 200 µL volume capacity for each well.…”

Section: D Design and Printing Of Pla Scaffoldsmentioning

confidence: 99%

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3D Printed Polylactic Acid (PLA) Well Plates for Enzyme Inhibition Studies: The Case of Pancreatic Lipase

STAMATIS¹,

Gournis²,

POLYDERA³

et al. 2022

Catal Res

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3D printed PLA has already been demonstrated for several biotechnological applications, including enzymes immobilization. The prerequisites for an efficient screening assay include using small volumes of reagents, low cost, and rapid screening of large numbers of compounds and extracts. Hence, assays based on microtiter plates are predominant. Thus, designing and fabricating scaffolds on a similar scale, which could serve as immobilization carriers, and their recruitment in inhibitors screening studies is of great significance, adding both enzyme stability and reuse potentiality of the biocatalytic system in assay merits. In this work, pancreatic lipase was immobilized on 3D-printed PLA microwells for enzyme inhibitor screening. XPS analysis demonstrated the successful modification of the PLA scaffolds. The immobilized enzyme displayed high levels of operational, thermal, and storage stability under the tested conditions. The IC<sub>50</sub> values for PPL inhibition were calculated for Orlistat, a model lipase inhibitor, and olive leaf extract, a promising natural compound. This is the first study reporting the use of 3D-printed PLA wells with an immobilized enzyme for inhibitor screening assay.

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Section: D Design and Printing Of Pla Scaffoldsmentioning

confidence: 99%

“…A synopsis of this process is presented in Figure 2. After designing and printing the desired model, the surfaces of the 3D scaffolds were modified according to our previous work [30]. The lipase from the porcine pancreas was immobilized inside the modified PLA wells with non-covalent interactions.…”

Section: D Printed Scaffolds For Ppl Inhibition Studiesmentioning

confidence: 99%

3D Printed Polylactic Acid (PLA) Well Plates for Enzyme Inhibition Studies: The Case of Pancreatic Lipase

STAMATIS¹,

Gournis²,

POLYDERA³

et al. 2022

Catal Res

Self Cite

View full text Add to dashboard Cite

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“…However, since cost reduction is one of the most important considerations in industry, studies were focused on the development of systems that can perform more than one reaction at the same time [37–40] . Therefore, more recently many studies concentrated on performing sequential enzymatic reactions by immobilizing more than one enzyme at the same time [37–40] .…”

Section: Introductionmentioning

confidence: 99%

“…[28,[34][35][36] However, since cost reduction is one of the most important considerations in industry, studies were focused on the development of systems that can perform more than one reaction at the same time. [37][38][39][40] Therefore, more recently many studies concentrated on performing sequential enzymatic reactions by immobilizing more than one enzyme at the same time. [37][38][39][40] However, it is important not only to carry out two enzymatic reactions at the same time or one after the other but also to develop a single system to eliminate by-products that may be released from enzymatic reactions and cause disadvantages.…”

Section: Introductionmentioning

confidence: 99%

“…[37][38][39][40] Therefore, more recently many studies concentrated on performing sequential enzymatic reactions by immobilizing more than one enzyme at the same time. [37][38][39][40] However, it is important not only to carry out two enzymatic reactions at the same time or one after the other but also to develop a single system to eliminate by-products that may be released from enzymatic reactions and cause disadvantages.…”

Section: Introductionmentioning

confidence: 99%

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Alpha‐Glucosidase Enzyme Immobilized Dextran‐Co Metal Nanoparticle Cryogel Composite Systems as Dual Catalyst with Enhanced Stability and Catalytic Activity

Demirci

Şahiner

2023

ChemCatChem

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Here, dextran (Dex) cryogels were prepared and used for the immobilization of α‐Glu enzyme via two different techniques of entrapment and covalent linking. The entrapment of α‐Glu enzyme within Dex cryogels (α‐GluE@Dex) during the synthesis process caused almost 70 % activity loss. On the other hand, the Dex cryogels with α‐Glu enzyme immobilized via covalent linking (α‐GluC@Dex) maintained 91±1.1 % activity and afforded 10 consecutive usages and could be stored at 25 °C for 10 days with more than 50 % activity. Moreover, we prepared Cobalt (Co) metal nanoparticle containing Dex‐Co cryogel composite to be used to immobilize the α‐Glu enzyme via covalent linking to attain an α‐GluC@Dex‐Co catalyst system with dual catalytic performance e. g., enzymatic and chemical reduction of the substrate and its’ by‐product for the first time. The prepared α‐GluC@Dex‐Co cryogels system was used for enzymatic hydrolysis of 4‐nitrophenyl‐α‐D‐glucopyranoside to glucose and 4‐nitrophenol, and then the reduction of 4‐nitrophenol to 4‐aminophenol catalyzed by the Co metal nanoparticles present as co‐catalyst within the cryogel network. The enzymatic activity of α‐GluC@Dex‐Co cryogels was calculated as 72±3.1 % and the cryogel reduced the enzymatic reaction by‐product of 4‐nitrophenol to 4‐aminophenol in 7 min in the presence of NaBH4.

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Development of 3D Printed Enzymatic Microreactors for Lipase-Catalyzed Reactions in Deep Eutectic Solvent-Based Media

et al. 2022

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In this study, 3D printing technology was exploited for the development of immobilized enzyme microreactors that could be used for biocatalytic processes in Deep Eutectic Solvent (DES)-based media. 3D-printed polylactic acid (PLA) microwell plates or tubular microfluidic reactors were modified with polyethylenimine (PEI) and lipase from Candida antarctica (CALB) was covalently immobilized in the interior of each structure. DESs were found to have a negligible effect on the activity and stability of CALB, and the system proved highly stable and reusable in the presence of DESs for the hydrolysis of p-nitrophenyl butyrate (p-NPB). A kinetic study under flow conditions revealed an enhancement of substrate accessibility in the presence of Betaine: Glycerol (Bet:Gly) DES, while the system was not severely affected by diffusion limitations. Incubation of microreactors in 100% Bet:Gly preserved the enzyme activity by 53% for 30 days of storage at 60 °C, while the buffer-stored sample had already been deactivated. The microfluidic enzyme reactor was efficiently used for the trans-esterification of ethyl ferulate (EF) with glycerol towards the production of glyceryl ferulate (GF), known for its antioxidant potential. The biocatalytic process under continuous flow conditions exhibited 23 times higher productivity than the batch reaction system. This study featured an effective and robust biocatalytic system with immobilized lipase that can be used both in hydrolytic and synthetic applications, while further optimization is expected to upgrade the microreactor system performance.

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3D Printed Polylactic Acid Well-Plate for Multi-enzyme Immobilization

Cited by 3 publications

References 10 publications

3D Printed Polylactic Acid (PLA) Well Plates for Enzyme Inhibition Studies: The Case of Pancreatic Lipase

3D Printed Polylactic Acid (PLA) Well Plates for Enzyme Inhibition Studies: The Case of Pancreatic Lipase

Alpha‐Glucosidase Enzyme Immobilized Dextran‐Co Metal Nanoparticle Cryogel Composite Systems as Dual Catalyst with Enhanced Stability and Catalytic Activity

Development of 3D Printed Enzymatic Microreactors for Lipase-Catalyzed Reactions in Deep Eutectic Solvent-Based Media

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