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      Novozym 435: the “perfect” lipase immobilized biocatalyst?

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          Abstract

          Novozym 435 (N435) is a commercially available immobilized lipase produced by Novozymes with its advantages and drawbacks.

          Abstract

          Novozym 435 (N435) is a commercially available immobilized lipase produced by Novozymes. It is based on immobilization via interfacial activation of lipase B from Candida antarctica on a resin, Lewatit VP OC 1600. This resin is a macroporous support formed by poly(methyl methacrylate) crosslinked with divinylbenzene. N435 is perhaps the most widely used commercial biocatalyst in both academy and industry. Here, we review some of the success stories of N435 (in chemistry, energy and lipid manipulation), but we focus on some of the problems that the use of this biocatalyst may generate. Some of these problems are just based on the mechanism of immobilization (interfacial activation) that may facilitate enzyme desorption under certain conditions. Other problems are specific to the support: mechanical fragility, moderate hydrophilicity that permits the accumulation of hydrophilic compounds ( e.g., water or glycerin) and the most critical one, support dissolution in some organic media. Finally, some solutions (N435 coating with silicone, enzyme physical or chemical crosslinking, and use of alternative supports) are proposed. However, the N435 history, even with these problems, may continue in the coming future due to its very good properties if some simpler alternative biocatalysts are not developed.

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          Most cited references502

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          Green chemistry: principles and practice.

          Green Chemistry is a relatively new emerging field that strives to work at the molecular level to achieve sustainability. The field has received widespread interest in the past decade due to its ability to harness chemical innovation to meet environmental and economic goals simultaneously. Green Chemistry has a framework of a cohesive set of Twelve Principles, which have been systematically surveyed in this critical review. This article covers the concepts of design and the scientific philosophy of Green Chemistry with a set of illustrative examples. Future trends in Green Chemistry are discussed with the challenge of using the Principles as a cohesive design system (93 references).
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            FAT SIGNALS - Lipases and Lipolysis in Lipid Metabolism and Signaling

            Lipolysis is defined as the catabolism of triacylglycerols stored in cellular lipid droplets. Recent discoveries of essential lipolytic enzymes and characterization of numerous regulatory proteins and mechanisms have fundamentally changed our perception of lipolysis and its impact on cellular metabolism. New findings that lipolytic products and intermediates participate in cellular signaling processes and that “lipolytic signaling” is particularly important in many nonadipose tissues unveil a previously underappreciated aspect of lipolysis, which may be relevant for human disease.
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              Modifying enzyme activity and selectivity by immobilization.

              Immobilization of enzymes may produce alterations in their observed activity, specificity or selectivity. Although in many cases an impoverishment of the enzyme properties is observed upon immobilization (caused by the distortion of the enzyme due to the interaction with the support) in some instances such properties may be enhanced by this immobilization. These alterations in enzyme properties are sometimes associated with changes in the enzyme structure. Occasionally, these variations will be positive. For example, they may be related to the stabilization of a hyperactivated form of the enzyme, like in the case of lipases immobilized on hydrophobic supports via interfacial activation. In some other instances, these improvements will be just a consequence of random modifications in the enzyme properties that in some reactions will be positive while in others may be negative. For this reason, the preparation of a library of biocatalysts as broad as possible may be a key turning point to find an immobilized biocatalyst with improved properties when compared to the free enzyme. Immobilized enzymes will be dispersed on the support surface and aggregation will no longer be possible, while the free enzyme may suffer aggregation, which greatly decreases enzyme activity. Moreover, enzyme rigidification may lead to preservation of the enzyme properties under drastic conditions in which the enzyme tends to become distorted thus decreasing its activity. Furthermore, immobilization of enzymes on a support, mainly on a porous support, may in many cases also have a positive impact on the observed enzyme behavior, not really related to structural changes. For example, the promotion of diffusional problems (e.g., pH gradients, substrate or product gradients), partition (towards or away from the enzyme environment, for substrate or products), or the blocking of some areas (e.g., reducing inhibitions) may greatly improve enzyme performance. Thus, in this tutorial review, we will try to list and explain some of the main reasons that may produce an improvement in enzyme activity, specificity or selectivity, either real or apparent, due to immobilization.
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                Author and article information

                Journal
                CSTAGD
                Catalysis Science & Technology
                Catal. Sci. Technol.
                Royal Society of Chemistry (RSC)
                2044-4753
                2044-4761
                May 20 2019
                2019
                : 9
                : 10
                : 2380-2420
                Affiliations
                [1 ]Escuela de Microbiología
                [2 ]Universidad Industrial de Santander
                [3 ]Bucaramanga
                [4 ]Colombia
                [5 ]Planta Piloto de Ingeniería Química – PLAPIQUI
                [6 ]CONICET
                [7 ]Universidad Nacional del Sur
                [8 ]8000 Bahía Blanca
                [9 ]Argentina
                [10 ]Departamento de Química
                [11 ]Facultad de Ciencias
                [12 ]Universidad del Tolima
                [13 ]Ibagué
                [14 ]Instituto de Engenharias e Desenvolvimento Sustentável
                [15 ]Universidade da Integração Internacional da Lusofonia Afro-Brasileira
                [16 ]Redenção
                [17 ]Brazil
                [18 ]Biotechnology, Bioprocess, and Biocatalysis Group, Food Science and Technology Institute
                [19 ]Federal University of Rio Grande do Sul
                [20 ]Porto Alegre
                [21 ]Instituto Universitario de Materiales
                [22 ]Departamento de Química Inorgánica
                [23 ]Universidad de Alicante
                [24 ]Alicante
                [25 ]Spain
                [26 ]Centro de Investigación y Desarrollo en Ciencias Aplicadas-Dr. Jorge J. Ronco
                [27 ]Universidad Nacional de La Plata
                [28 ]Buenos Aires
                [29 ]Department of Biocatalysis
                [30 ]ICP-CSIC
                [31 ]Campus UAM-CSIC
                [32 ]Cantoblanco
                [33 ]Madrid
                Article
                10.1039/C9CY00415G
                488d05a9-878a-445b-8cfa-e8318b7ee4fe
                © 2019

                http://creativecommons.org/licenses/by-nc/3.0/

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