This work reports the use of the Octave, a free computational software, to optimize the experimental design in the synthesis of ethyl oleate from oleic acid and ethanol catalyzed by a native lipase from Aspergillus niger. The central composite factorial design (CCD) was performed with 2 levels and 3 variables (temperature, mass of native A. niger lipase and oleic acid:ethanol molar ratio) in conjunction with response surface methodology (RSM). The results indicate that variable 1 (oleic acid:ethanol molar ratio; X1) and 2 (mass of A. niger lipase, X2), as well as the interaction between them, are statistically significant. The optimal reaction condition was estimated using ANOVA data and the response surface. The experimental results are consistent with those estimated, and thus, ethyl oleate was obtained with conversion (> 90%) using oleic acid and ethanol in a 1:2 molar ratio, A. niger lipase (8,00 mg) and temperature of 45 °C in 6 hours of reaction.

O desempenho de duas lipases nativas (lipase de Aspergillus niger e Rhizopus oligosporus), e a influência da temperatura, doador acila (acetato de etila, acetato de vinila, acetato de iso-propenila e anidrido acético) e do meio orgânico foi avaliado na resolução da (RS)-feniletilamina (1). O efeito de anions em uma série de líquidos iônicos (LIs) baseados em derivados do imidazol [BMIm][X], onde X = BF4, PF6, SCN e Cl em n-heptano e acetato de vinila, também foi verificado com a lipase nativa de A. niger. Com esta lipase, a amida R-2b foi obtida com conversões de 6 até > 99% e valores de E de 2 até > 200, em n-heptano ou n-hexano. Os maiores valores de E obtidos, usando sistema bifásico formado por n-heptano e [BMIm][ PF6] ou [BMIm][ BF4] 9:1 (v/v), foram de 9 e 7, respectivamente, quando comparado com o solvente puro (E = 2). A influência dos ânions segue a seguinte ordem: PF6- >BF4- > SCN- > Cl-.

The performance of two native lipases (lipase from Aspergillus niger and Rhizoupus oligosporus) in the resolution of (RS)-phenylethylamine (1), varying the temperature, acyl donor type (ethyl acetate, vinyl acetate, iso-propenyl acetate and acetic anhydride) and organic medium, was studied. The effect of the nature of the anion using native A. niger lipase in n-heptane with a series of imidazolium-based ILs [BMIm][X], where X = BF4, PF6, SCN and Cl, was also evaluated. Using the lipase from A. niger, the R-2b amide was obtained with conversions from 6 to > 99% and E-values from 2 to > 200, with n-heptane or n-hexane. This lipase showed better E-values in a two-phase system using n-heptane and [BMIm][PF6] or [BMIm][BF4] 9:1 (v/v), obtaining values of 9 and 7, respectively, when vinyl acetate was used as the acyl donor, compared to the use of pure n-heptane (E = 2). The series for the anions in terms of decreasing performance was as follows: PF6- >BF4- > SCN- > Cl-.

Três lipases microbianas nativas (Aspergillus niger, Rhizopus javanicus e Penicillium solitum) foram utilizadas na hidrólise do óleo de salmão (teor de AGPI n-3 de 30,1%) com o objetivo de concentrar o conteúdo de ácidos graxos poliinsaturados n-3 (AGPI n-3) nos acilgliceróis residuais. A metodologia de planejamento experimental e análise de superfície de resposta foi usada para se chegar às condições otimizadas de cada reação enzimática, utilizando as seguintes variáveis; temperatura (X1), quantidade de lipase (X2) e taxa de água/óleo (X3). Com base nos resultados do planejamento, a lipase de Aspergillus niger foi a mais eficiente na concentração dos AGPI n-3, sendo que as condições ótimas de reação foram: concentração de enzima de 500 U g-1 óleo, temperatura 45 ºC e taxa de água/óleo de 2:1 m/m após 24 h de reação. O grau de hidrólise (60%) conduziu a um aumento do conteúdo de ácido docosahexaenóico (DHA) de 14,4% para 34,0% (enriquecimento de 2,4 vezes) nos acilgliceróis residuais após a hidrólise do óleo de salmão.

In an attempt to concentrate the content of n-3 polyunsaturated fatty acids (n-3 PUFA) in the residual acylglycerol, salmon oil (n-3 PUFA content of 30.1%) was hydrolyzed with three kinds of native microbial lipases (Aspergillus niger, Rhizopus javanicus and Penicillium solitum). For each lipase, a response surface methodology was used to obtain maximum PUFA content and to optimize the parameters of enzymatic reactions with respect to important reaction variables; temperature (X1), amount of lipases (X2) and water/oil ratio (X3). Based on these results, optimal reaction conditions were established. Aspergillus niger lipase was the most effective in concentrating n-3 PUFA. The degree of hydrolysis (60%) led to an increase in the docosahexaenoic acid (DHA) content from 14.4% in the original oil to 34.0% (2.4-fold enrichment) in the residual acylglycerol under optimum conditions: enzyme concentration of 500 U g-1 oil, reaction temperature of 45 ºC and water/oil mass rate of 2:1 (m/m) after 24 h reaction.

Lipase de Aspergillus niger foi imobilizada por deposição utilizando Celite e as atividades de hidrólise e esterificação, a estabilidade e a enantioseletividade das formas livre e imobilizada foram comparadas. Ambas as preparações mostraram propriedades bioquímicas similares, com atividade máxima em pH 6,0 e temperatura de 30-40 ºC. Não foram observadas diferenças na atividade hidrolítica em função do pH para as duas formas da enzima. Os efeitos mais importantes observados após a imobilização foram a estabilidade térmica e a melhoria da atividade de esterificação na reação do (R,S)-ibuprofeno com 1-propanol em presença do isooctano. Além disso, a lipase na forma imobilizada manteve, pelo menos, 73% de sua atividade de esterificação após 5 dias de armazenamento a 40 ºC e pôde ser reciclada e reutilizada por pelo menos 6 vezes.

Aspergillus niger lipase was immobilized by deposition on Celite and the hydrolytic and esterification activities, stability and enantioselectivity of both free and immobilized enzyme were compared. Both the free and immobilized preparations showed similar biochemical properties, with maximum activity at pH 6.0 and a temperature of 30-40 ºC. The most important effects observed when the lipase was immobilized were thermal stability and an improved esterification activity during the reaction of (R,S)-ibuprofen with 1-propanol in isooctane. Moreover, immobilized Aspergillus niger lipase maintained an esterification activity of at least 73% after 5 days of storage at 40 ºC and can be recycled and reused at least 6 times.

Microbial lipases have a great potential for commercial applications due to their stability, selectivity and broad substrate specificity because many non-natural acids, alcohols or amines can be used as the substrate. Three microbial lipases isolated from Brazilian soil samples (Aspergillus niger; Geotrichum candidum; Penicillium solitum) were compared in terms of their stability and as biocatalysts in the enantioselective esterification using racemic substrates in organic medium. The lipase from Aspergillus niger showed the highest activity (18.2 U/mL) and was highly thermostable, retaining 90% and 60% activity at 50 ºC and 60 ºC after 1 hour, respectively. In organic medium, this lipase provided the best results in terms of enantiomeric excess of the (S)-active acid (ee = 6.1%) and conversion value (c = 20%) in the esterification of (R,S)-ibuprofen with 1-propanol in isooctane. The esterification reaction of the racemic mixture of (R,S)-2-octanol with decanoic acid proceeded with high enantioselectivity when lipase from Aspergillus niger (E = 13.2) and commercial lipase from Candida antarctica (E = 20) were employed.