Adaptação do biorreator do tipo airlift para reação enzimática empregando lipase imobilizada® na síntese de lipídeos estruturados do tipo MLM
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Data
2021-06-10
Autores
Oliveira, Daniele Gonçalves de
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Universidade Estadual Paulista (Unesp)
Resumo
As indústrias de alimentos e fármacos incorporam cada vez mais tecnologia em seus produtos, como a modificação de óleos e gorduras para a obtenção de triglicerídeos estruturados. Desta forma, cresce o interesse por biorreatores para sistemas trifásicos como os que utilizam enzimas imobilizadas. Os biorreatores airlift se mostram atraentes para este tipo de processo, pois fornecem uma mistura controlada, baixa taxa de cisalhamento e uma eficiente suspensão de sólidos. Logo, o objetivo do trabalho foi a adaptação de um biorreator airlift de tubos concêntricos usualmente empregado em fermentação microbiana para a síntese enzimática de lipídeos estruturados, especificamente os triglicerídeos dietéticos do tipo MLM, isto é, aqueles que possuem ácidos graxos de cadeia média (M), nas posições sn-1 e sn-3, e de cadeia longa (L), na posição sn-2 da molécula de triacilglicerol. Foi selecionado o óleo de semente de uva, com base em sua composição rica em ácidos graxos insaturados. Este óleo foi submetido a reações de acidólise com o ácido graxo de cadeia média ácido cáprico (C10:0), tal reação foi catalisada pela lipase imobilizada, Lipozyme RM IM ® (Rhizomucor miehei). Primeiramente, com o auxílio de uma impressora 3D confeccionou-se um tubo interno com estrutura de um tronco de um cone e um fundo abaulado, objetivando a otimização da mistura no biorreator. Após, caracterizou-se sua hidrodinâmica, obtendo melhor tempo de mistura na vazão de ar de 2 vvm para as três cargas de sólidos (suporte de imobilização da enzima) estudadas. Posteriormente, realizou-se reações de acidólise em batelada variando a vazão de ar em 1 e 2 vvm utilizando 1% (v/v) de carga de enzimas, obtendo graus de incorporação (GI) de C10 igual 24,02± 0,13% para 1 vvm e 28,76 ± 0,05% para reação à 2 vvm. Realizaram-se reações com 2 e 3 %(v/v) de enzimas imobilizadas e observou-se que com o aumento de carga enzimática ocorre aceleração da velocidade de incorporação de ácidos graxos de cadeia média .O maior grau de incorporação obtido foi na reação com maior massa enzimática, 3% (v/v) em relação ao meio reacional no tempo 24h, 32,32 ± 1,1%. Outro ponto observado foi que o aumento do índice de peróxido durante esse período foi desprezível, pois houve um aumento de 0,72±0,10 para 2,82±0,28 mEq/Kg estando abaixo do estipulado pela ANVISA que é 10mEq/Kg. Tais resultados, mostram que o biorreator do tipo airlift é apto a realizar misturas multifásicas, mostrando-se um excelente reator para reações enzimáticas, especialmente para a síntese de triglicerídeos dietéticos do tipo MLM.
The food and pharmaceutical industries are increasingly incorporating technology into their products, such as modifying oils and fats to obtain structured triglycerides. Thus, interest in bioreactors for three-phase systems such as those using immobilized enzymes grows. Airlift bioreactors are attractive for this type of process, as they provide a controlled mixing, low shear rate, and efficient suspension of solids. Therefore, the objective of the work was the adaptation of a concentric tube airlift bioreactor usually used in microbial fermentation for the enzymatic synthesis of structured lipids, specifically MLM-type dietary triglycerides, that is, those with medium-chain fatty acids (M ), at the sn-1 and sn-3 positions, and long-chain (L), at the sn-2 position of the triacylglycerol molecule. Grapeseed oil was selected, based on its rich composition in unsaturated fatty acids. This oil was subjected to acidolysis reactions with the medium-chain fatty acid capric acid (C10:0), this reaction was catalyzed by the immobilized lipase, Lipozyme RM IM ® (Rhizomucor miehei). First, with the aid of a 3D printer, an inner tube was made with a trunk of a cone structure and a convex bottom, aiming at optimizing the mixture in the bioreactor. Afterward, its hydrodynamics were characterized, obtaining the best mixing time at an airflow of 2 vvm for the three solid loads (enzyme immobilization support) studied. Subsequently, batch acidolysis reactions were carried out, varying the airflow in 1 and 2 vvm using 1% (v/v) enzyme load, obtaining degrees of incorporation (GI) of C10 equal to 24.02 ± 0.13 % for 1 vvm and 28.76 ± 0.05% for reaction to 2 vvm. Reactions were carried out with 2 and 3%(v/v) of immobilized enzymes and it was observed that with the increase in the enzymatic load, there was an acceleration of the incorporation rate of medium-chain fatty acids. The highest degree of incorporation obtained was in the reaction with greater enzymatic mass, 3% (v/v) in relation to the reaction medium in time 24h, 32.32 ± 1.1%. Another point observed was that the increase in the peroxide index during this period was negligible, as there was an increase from 0.72±0.10 to 2.82±0.28 mEq/Kg, being below the stipulated by ANVISA, which is 10mEq/ Kg These results show that the airlift-type bioreactor is able to carry out multiphase mixtures, proving to be an excellent reactor for enzymatic reactions, especially for the synthesis of MLM-type dietary triglycerides.
The food and pharmaceutical industries are increasingly incorporating technology into their products, such as modifying oils and fats to obtain structured triglycerides. Thus, interest in bioreactors for three-phase systems such as those using immobilized enzymes grows. Airlift bioreactors are attractive for this type of process, as they provide a controlled mixing, low shear rate, and efficient suspension of solids. Therefore, the objective of the work was the adaptation of a concentric tube airlift bioreactor usually used in microbial fermentation for the enzymatic synthesis of structured lipids, specifically MLM-type dietary triglycerides, that is, those with medium-chain fatty acids (M ), at the sn-1 and sn-3 positions, and long-chain (L), at the sn-2 position of the triacylglycerol molecule. Grapeseed oil was selected, based on its rich composition in unsaturated fatty acids. This oil was subjected to acidolysis reactions with the medium-chain fatty acid capric acid (C10:0), this reaction was catalyzed by the immobilized lipase, Lipozyme RM IM ® (Rhizomucor miehei). First, with the aid of a 3D printer, an inner tube was made with a trunk of a cone structure and a convex bottom, aiming at optimizing the mixture in the bioreactor. Afterward, its hydrodynamics were characterized, obtaining the best mixing time at an airflow of 2 vvm for the three solid loads (enzyme immobilization support) studied. Subsequently, batch acidolysis reactions were carried out, varying the airflow in 1 and 2 vvm using 1% (v/v) enzyme load, obtaining degrees of incorporation (GI) of C10 equal to 24.02 ± 0.13 % for 1 vvm and 28.76 ± 0.05% for reaction to 2 vvm. Reactions were carried out with 2 and 3%(v/v) of immobilized enzymes and it was observed that with the increase in the enzymatic load, there was an acceleration of the incorporation rate of medium-chain fatty acids. The highest degree of incorporation obtained was in the reaction with greater enzymatic mass, 3% (v/v) in relation to the reaction medium in time 24h, 32.32 ± 1.1%. Another point observed was that the increase in the peroxide index during this period was negligible, as there was an increase from 0.72±0.10 to 2.82±0.28 mEq/Kg, being below the stipulated by ANVISA, which is 10mEq/ Kg These results show that the airlift-type bioreactor is able to carry out multiphase mixtures, proving to be an excellent reactor for enzymatic reactions, especially for the synthesis of MLM-type dietary triglycerides.
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Palavras-chave
Reator airlift, Triglicerídeos dietéticos, Acidólise, Lipase®, Óleo de semente de uva, airlift reactor, Dietary triglycerides, Acidolysis, Grape seed oil