Immobilization of Yarrowia lipolytica Lipase—A Comparison of Stability of Physical Adsorption and Covalent Attachment Techniques

Cunha, Aline G.; Fernández‐Lorente, Gloria; Bevilaqua, Juliana V.; Destain, Jacqueline; Paiva, Lucia Moreira Campos; Freire, Denise M. G.; Fernández‐Lafuente, Roberto; Guisán, José M.

doi:10.1007/978-1-60327-526-2_19

Cited by 7 publications

(7 citation statements)

References 25 publications

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“…On this last point, benefits of enzyme immobilization include: facilitating the separation of products from the active enzymes; and increasing enzyme shelf life by protecting them against proteolysis, thermal and chemical denaturation. [42][43][44] There are several methods for the industrial immobilization of enzymes, such as: covalent linkage 44 or adsorption 45 (generally ionic) to a support; entrapment within a matrix; 46 and direct enzyme crosslinking. 47 There are disadvantages of these current methods, however, which include: the leakage of the protein from supports; loss of enzyme activity due to protein unfolding or misfolding; and restricted protein mobility and/or access of substrates due to inappropriate cross-linking.…”

Section: Resultsmentioning

confidence: 99%

Decorating Self-Assembled Peptide Cages with Proteins

et al. 2017

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An ability to organize and encapsulate multiple active proteins into defined objects and spaces at the nanoscale has potential applications in biotechnology, nanotechnology, and synthetic biology. Previously, we have described the design, assembly, and characterization of peptide-based self-assembled cages (SAGEs). These ≈100 nm particles comprise thousands of copies of de novo designed peptide-based hubs that array into a hexagonal network and close to give caged structures. Here, we show that, when fused to the designed peptides, various natural proteins can be co-assembled into SAGE particles. We call these constructs pSAGE for protein-SAGE. These particles tolerate the incorporation of multiple copies of folded proteins fused to either the N or the C termini of the hubs, which modeling indicates form the external and internal surfaces of the particles, respectively. Up to 15% of the hubs can be functionalized without compromising the integrity of the pSAGEs. This corresponds to hundreds of copies giving mM local concentrations of protein in the particles. Moreover, and illustrating the modularity of the SAGE system, we show that multiple different proteins can be assembled simultaneously into the same particle. As the peptide-protein fusions are made via recombinant expression of synthetic genes, we envisage that pSAGE systems could be developed modularly to actively encapsulate or to present a wide variety of functional proteins, allowing them to be developed as nanoreactors through the immobilization of enzyme cascades or as vehicles for presenting whole antigenic proteins as synthetic vaccine platforms.

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Section: Resultsmentioning

confidence: 99%

Decorating Self-Assembled Peptide Cages with Proteins

et al. 2017

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“…These results indicated that relatively weaker hydrophobic environment did not contribute to the improvement of specific activity of immobilized lipase. The advantages of PGMA were mainly due to the covalent attachment between lipase and the microspheres [22,28,36]. Catalytic efficiency, reusability, and stability of immobilized lipase…”

Section: Influence Of Different Hydrophobicity Degree On Lipase Immobmentioning

confidence: 99%

“…Therefore, many researchers focused on covalent binding of lipase molecules with the functional groups of carriers to obtain stable enzyme performance [21][22][23]. The functional groups of carriers mainly include epoxy [24,25], amide [26], and aldehyde group [27].…”

Section: Introductionmentioning

confidence: 99%

Comparison of covalent and physical immobilization of lipase in gigaporous polymeric microspheres

Wang

Zhou

et al. 2015

Bioprocess Biosyst Eng

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Lipase (EC 3.1.1.3) is a versatile enzyme which has been widely used in ester-reaction industries. We have previously discovered that gigaporous polystyrene (PST) microspheres can be used as a novel immobilization carrier for lipase. In this work, a series of gigaporous microspheres with different densities of epoxy group including poly(glycidyl methacrylate) (PGMA) and poly(styrene-co-glycidyl methacrylate) [P(ST-GMA)] were evaluated as lipase immobilization carriers, which were also compared with gigaporous PST microspheres and the commercial immobilized lipase Novozym 435. Lipase immobilized in gigaporous PGMA microspheres showed the highest activity yield, reusability, and stability as well as the best affinity for the substrate. The characterizations of adsorption curves, the change of epoxy group amounts, and hydrophobic-hydrophilic properties of the microspheres were carried out to investigate the interaction between lipase molecules and carriers. It was found that covalent binding played a key role in improving the properties of lipase immobilized in gigaporous PGMA microspheres.

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“…Many diverse immobilization methods have been developed for this purpose. Among these are some methods that require rigid supports: most notably, covalent immobilization [9,11,17], ionic immobilization [20], and hydrophobic adsorption [2]. However, methods for the immobilization of enzymes without the use of supports are gaining in importance because they offer the advantages of high volumetric productivity and lower production costs [16,17].…”

mentioning

confidence: 99%

“…Here, lipase enzymes could be applied for biodiesel production in an eco-friendly manner [14]. For decades, much of the literature relevant to this topic has been focused largely on the process of biodiesel production using microbial lipases [2,9,11,20]. Among these, Candida antarctica lipase B (CalB) has been most frequently utilized, owing primarily to its profound ability to carry out the biodiesel production reaction with a variety of plant oils.…”

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confidence: 99%

Transesterification Using the Cross-Linked Enzyme Aggregate of Photobacterium lipolyticum Lipase M37

Han

Kim²

2011

J. Microbiol. Biotechnol.

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Biodiesel is methyl and ethyl esters of long-chain fatty acids produced from vegetable oils or animal fats. Lipase enzymes have occasionally been used for the production of this biofuel. Recently, biodiesel production using immobilized lipase has received increased attention. Through enhanced stability and reusability, immobilized lipase can contribute to the reduction of the costs inherent to biodiesel production. In this study, methanol-tolerant lipase M37 from Photobacterium lipolyticum was immobilized using the cross-linked enzyme aggregate (CLEA) method. Lipase M37 has a high lysine content (9.7%) in its protein sequence. Most lysine residues are located evenly over the surface of the protein, except for the lid structure region, which makes the CLEA preparation yield quite high (~93%). CLEA M37 evidences an optimal temperature of 30 o C, and an optimal pH of 9-10. It was stable up to 50 o C and in a pH range of 4.0-11.0. Both soluble M37 and CLEA M37 were stable in the presence of high concentrations of methanol, ethanol, 1-propanol, and n-butanol. That is, their activities were maintained at solvent concentrations above 10% (v/v). CLEA M37 could produce biodiesel from olive oil and alcohols such as methanol and ethanol. Additionally, CLEA M37 generated biodiesel via both 2-step methanol feeding procedures. Considering its physical stability and reusability, CLEA M37 may potentially be used as a catalyst in organic synthesis, including the biodiesel production reaction.

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Immobilization of Yarrowia lipolytica Lipase—A Comparison of Stability of Physical Adsorption and Covalent Attachment Techniques

Cited by 7 publications

References 25 publications

Decorating Self-Assembled Peptide Cages with Proteins

Decorating Self-Assembled Peptide Cages with Proteins

Comparison of covalent and physical immobilization of lipase in gigaporous polymeric microspheres

Transesterification Using the Cross-Linked Enzyme Aggregate of Photobacterium lipolyticum Lipase M37

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