In this study, various commercial and native lipases, free or immobilized, were used in the enantioselective resolution of (R,S)-menthol with vinyl acetate for the preparation of menthyl acetate. Lipase from Candida rugosa (AYL) was found to be the most appropriate. When the reaction was carried out at 35 ºC, using 40 mg of AYL and a molar ratio of 1:2 (alcohol:ester), the values for the conversion degree, the enantiomeric excess of the substrate (ees) and product (eep) and the enantiomeric ratio (E) were 33.6, 50.3, > 99% and > 200, respectively. The results showed that the reaction was more efficient using toluene in comparison with other organic solvents or mixtures of toluene and ionic liquids (ILs). The best support for the AYL immobilization, was corn starch film. This system was used 4 times over 71 days of storage, and menthyl acetate was still obtained with good values of conversion degree, ees, eep, and E.

The immobilization of enzymes and microorganisms on solid supports has been developed in recent years. These biocatalysts may be used in organic media allowing their storage and reuse, thus reducing costs of the process. Herein, lipases from various sources were immobilized in agar gel and used as catalysts in the chemo-enzymatic epoxidation of β-caryophyllene. Several experimental parameters, such as the use of different organic solvents including ionic liquids, time, temperature, and agitation rate were evaluated. The mono-epoxide was obtained as a single product. The best result was achieved using immobilized F-AP15 lipase, forming the corresponding β-caryophyllene epoxide at a conversion of 96% in an 8h reaction at 35 ºC.

As lipases CAL-B, LPS, LPS-C, LPS-D e a de A. níger, livres ou imobilizadas em filmes de amido de cará (Discorea alata L.), inhame (Colocasia esculenta (L.) Schott) ou de gelatina, foram utilizadas na acilação da (RS)-sec-butilamina com diferentes doadores acila em vários solventes orgânicos sob aquecimento convencional (CH) ou irradiação por microondas (MW). Ao utilizar a lipase de A. niger livre, a conversão foi três vezes maior sob irradiação por MW em relação ao CH a 35 ºC. Usando a lipase de A. niger livre, a (R)-amida foi obtida com 21% de conversão, resultando em valores de ee p > 99% e de E (enantiosseletividade) > 200, em 1 min de reação em MW. Ao utilizar a lipase de A. niger imobilizada em filmes de amido de cará e inhame, em 3 ou 5 min de reação em MW, a amida foi obtida com conversões moderadas, sendo de 8 a 25%, porém com maior seletividade (ee p > 99% e E > 200) em comparação à forma livre (45% de conversão, ee p 81% e 18 para o valor de E).

The lipases CAL-B, PSL, PSL-C, PSL-D, and A. niger lipase, free or immobilized in starch (obtained from two types of yam, known in Brazil as “cará” (Discorea alata L.) and “inhame” (Colocasia esculenta (L.) Schott) or gelatin films, were used in the acylation of (RS)-sec-butylamine with different acyl donors in various organic solvents applying conventional heating (CH) or microwave (MW) irradiation. In the case of free A. niger lipase, the conversion degrees were three times higher using MW irradiation when compared to conventional heating at 35 ºC. Using free A. niger lipase, the (R)-amide was obtained with a conversion degree of 21%, resulting in ee p > 99% and E-value (enantioselectivity value) > 200, in 1 min of reaction under MW irradiation. When the A. niger lipase was immobilized in yam starch films, the (R)-amide was obtained in moderate conversions of 8-25% after 3 or 5 min of reaction under MW irradiation, but with higher selectivity (ee p > 99% and E > 200) in comparison with the free form (conversion degree of 45%, ee p 81% and E value of 18).

In recent years, the introduction of the Green Chemistry concepts in undergraduate chemistry classes has been intensively pursued. In this regard, the two-step preparation of Epoxone (an organocatalyst developed by Shi & col.) from commercial D-fructose, through ketalization of vicinal diols followed by oxidation of a sterically congested secondary alcohol, involves important topics in Organic Chemistry and employs inexpensive and nontoxic reagents. The reactions are easy to perform and the products from both steps are readily obtained as crystalline solids after simple procedures, thus facilitating their chemical characterization.

Foi realizada a epoxidação quimio-enzimática de ésteres metílicos do óleo de girassol, utilizando lipase em meio aquoso, na presença ou não de acil doadores. As reações no sistema bifásico (CH2Cl2/H2O) foram conduzidas utilizando-se a Candida antarctica lipase B (CALB, 1000 u g-1) e peróxido de hidrogênio a 30% (v/v). A conversão foi maior do que 99% em 16 h de reação. Os melhores resultados foram obtidos com o sistema bifásico, em tempo reacional de 16 h, a 30 ºC, 10 mmol de ácido octanóico em relação a 1 g de óleo, 6 mL de diclorometano e 5 mL de água, garantindo uma alta conversão dos ésteres e maior tempo de uso da lipase.

The chemo-enzymatic epoxidation of the methyl esters of sunflower oil with lipase from Candida antarctica B and aqueous H2O2 in the presence and absence of an acyl donor was investigated. The biphasic system (CH2Cl2/H2O) comprised Candida antarctica B lipase (CALB, 1000 u g-1) and 30% (v/v) aqueous hydrogen peroxide. In some cases the conversion was higher than 99%. The best results were obtained for the biphasic system after 16 h of reaction, at 30 ºC, using 10 mmol of octanoic acid in relation to 1 g of the oil, 6 mL of dichlomethane and 5 mL of water.

A doença de Newcastle é caracterizada por manifestações respiratórias associadas a sintomas nervosos e/ou digestivos. Sua prevenção é feita pela vacinação com vacinas vivas atenuadas (cepas lentogênicas) e/ou inativadas. As cepas lentogênicas podem determinar acentuada reação pós-vacinal, principalmente na presença de outros patógenos. Entre eles, o Mycoplasma synoviae tem importância mundial, principalmente no Brasil. A disseminação deste agente nos planteís avícolas tem sido facilitada, devido a dificuldades de reprodução e diagnóstico da doença em aves, variação de virulência entre as diferentes cepas de M.synoviae e atribuição a outros patógenos de manifestação típica da micoplasmose por M.synoviae. Este estudo experimental em aves (Gallus gallus) SPF, previamente infectadas por M.synoviae e depois vacinadas contra Newcastle, foi realizado com objetivo de avaliar a patogenicidade do M.synoviae pela obtenção dareação respiratória pós-vacinal e a resposta sorológica para o vírus vacinal da doença de Newcastle, na ausência de fatores ambientais. Um total de 86 aves, com três dias de idade foram utilizadas, sendo 57 infectadas via ocular e intranasal, com cepa MS WVU 1853, ativada em galinhas. Sete dias depois, 21 aves infectadas por micoplasma e 29 não infectadas foram vacinadas contra a doença de Newcastle. Como resultados, aves não infectadas e vacinadas produziram resposta sorológica para o vírus vacinal da doença de Newcastle, significativamente mais elevada e mais duradora que aquelas infectadas e vacinadas. Igualmente, aves infectadas e vacinadas produziram reações sorológicas para M.synoviae mais baixa, que aquelas apenas infectadas. Não foram observadas reações respiratórias pós-vacinal nas aves vacinadas.

Newcastle disease is characterized by respiratory manifestations in association with nervous and/or digestive symptoms. Its prevention is done by vaccination with live attenuated (lentogenic strains) and/or killed vaccines. The lentogenic strains can lead to strong post-vaccination reaction, principally due to the presence of other pathogenic agents. Among them, Mycoplasma synoviae is worldwide important, mainly in Brazil. The dissemination of this agent in poultry flocks has been achieved due to difficulties in diagnosis and disease reproduction, virulence variations among different M.synoviae strains, and attribution of typical M.synoviae disease manifestation to other disease agents. This experimental study in SPF chicks (Gallus gallus), previously infected by M.synoviae and thereafter vaccinated against Newcastle disease, was done with the objective of evaluating M.synoviae pathogenicity through assessment of post-vaccinal respiratory reactions and serologic responses to Newcastle disease virus vaccine in the absence of environmental factors. A total of 86 three days old chicks were used, being 57 infected by eye and nostril drop, with chicken activated M. synoviae strain WVU 1853. Seven days later, 21 mycoplasma infected birds plus 29 not mycoplasma infected ones were vaccinated against Newcastle disease. As results, the not infected and vaccinated birds yielded, significantly, higher and longer lasting serologic responses to Newcastle disease vaccine virus than those infected and vaccinated. Similarly, the infected and vaccinated birds yielded lower serologic reactions to M.synoviae than those only mycoplasma infected. No post-vaccinal respiratory reaction was observed in the vaccinated birds.

Lipases from different sources were immobilized in sodium caseinate/glycerol film and used in the esterification reactions of aliphatic acids with alcohols in the presence of organic solvents. Lipases from Pseudomonas sp and Rhizopus oryzae were selected and the influence of several parameters was analyzed, including: lipase loading, organic solvent polarity, reaction temperature, chain length of alcohol and acid and enzyme/support reuse. For comparison, free enzymes were used under similar experimental conditions.

The application of biocatalysis is a promising field related to new technologies for organic synthesis. The development of immobilization techniques is very important due to the multiple or repetitive use of a single batch of enzymes and the ability to stop the reaction rapidly, at any stage, by removing the enzymes. In most cases, after immobilization, enzymes and microorganisms maintain or even increase their activity and stability. This work presents an overview of the common methods for lipase immobilization in polymers and applications of these systems to obtain compounds of synthetic interest.

A series of seven Schiff bases have been synthesized from 3,3-diphenylpropilamine and substituted benzaldehydes. These imines were treated with NaBH4 in ethanol affording the corresponding amines in 98-55% yields. A molecular modeling study was performed with the Schiff bases in order to compare the theoretical parameters with the experimental results. The theoretical parameters were obtained by AM1 and PM3 semi-empirical methods. The analysis of charge, electron densities and LUMO coefficients suggested that the most favorable interactions should occur with Schiff bases containing electron-donating groups, in accordance with experimental yields, showing that the higher reactivity is due to higher electrophilic character of imine carbons.

O presente estudo foi desenvolvido em ratos de dois biotérios, um em Niterói e outro no Rio de Janeiro, sendo um classificado como convencional controlado e o outro como convencional. O objetivo foi verificar a presença do Mycoplasma pulmonis em animais que apresentavam sintomas de doença respiratória e baixa produtividade. No biotério convencional controlado foram necropsiados 16 ratos da linhagem Wistar-Furth enquanto no biotério convencional a necrópsia foi realizada em 12 ratos da linhagem Hooded Lister. Os espécimes de pulmão, traquéia, orofaringe, ouvido médio, útero e ovário foram submetidas ao cultivo e o soro obtido ao teste de ELISA. Dos 28 ratos que foram submetidos ao cultivo para micoplasmas, 57,14% (16/28) foram positivos para M. pulmonis sendo 81,25% (13/16) provenientes do biotério convencional controlado, enquanto no biotério convencional a taxa foi de 25,00% (3/12). O teste de ELISA foi realizado em soro de 20 animais, de ambos os biotérios, verificando-se que no biotério convencional controlado, 92,86% (13/14) dos ratos examinados, foram positivos enquanto 7,14% (1/14) foram considerados suspeitos. No biotério convencional, 100% (6/6) dos ratos foram positivos. Esses resultados confirmaram que M.pulmonis era o agente etiológico da doença respiratória que acometia os animais estudados e que a sensibilidade do ELISA foi maior do que a do isolamento.

This work was conducted on rats in two premises located in Niterói and Rio de Janeiro, Brazil. One is classified as conventional controlled and the other, conventional. The objective of the present study was to detect the presence of Mycoplasma pulmonis in animals with symptoms of respiratory disease and low reproductive performance. In the conventional controlled premises, 16 rats of Wistar-Furth strain were necropsied while in the conventional premises necropsy was performed on 12 rats of Hooded Lister strain. The clinical samples of lungs, trachea, oropharynx, middle ear, uterus and ovaries were subjected to culturing while the sera were tested for antibody detection. From 28 rats, 57.14% (16/28) were culture positive for M. pulmonis, being 81.25% (13/16) from the conventional controlled premises, and 25.00% (3/12) from the conventional premises. The ELISA test was carried out in 20 animals of both colonies. In the conventional controlled premises, 92.86% (13/14) were positive for M. pulmonis, and 7.14% (1/14) were suspicious, while in the conventional premises, 100% (6/6) of the samples were positive. The results confirmed that M. pulmonis was the etiologic agent of the disease that affected the rats under study, and that the ELISA positivity rated higher than culture.

New methodologies for protection of Saccharomyces cerevisiae (FP) cells when supported in montmorillonite K10 (K10), recovered or not with gelatin (G) and in the presence or absence of sucrose (S) are presented. These systems were used for the enantioselective reduction of ethyl acetoacetate and a-chloroacetophenone in hexane, under FP/K10/G/S and FP/S at 20ºC during 24 hours, affording S-(+)-ethyl-3-hydroxybutanoate in 100% conversion and 99% ee, and R-(-)-2-chloro-1-phenylethanol 79% and 78% ee at 20 and 30 ºC, respectivelly.

Uma série de Bases de Schiff foi sintetizada através da reação de condensação do piperonal (3,4-metilenodioxibenzaldeido) com as correspondentes aminas primárias aromáticas. Seus espectros de RMN de ¹H e 13C foram obtidos e estudou-se as correlações de Hammett envolvendo os deslocamentos químicos e as constantes dos substituíntes (sigmap, sigmaR e sigmaI). Observou-se correlações lineares e bilineares significativas para o carbono imínico (C-alfa) e C-1’, indicando maior contribuição do efeito de ressonância do substituinte sobre os deslocamentos químicos. Os assinalamentos dos C-4’ apresentaram-se bem alterados pelos efeitos dos substituíntes, especialmente pelos halogênios, na direção esperada. A série de Bases de Schiff foi ensaiada contra microorganismos, sendo que apresentou atividade significativa frente a fungos filamentosos, especialmente o Epidermophyton floccosum. Não foram observadas correlações significativas entre a atividade biológica e os efeitos eletrônicos.

A series of eleven Schiff Bases have been synthesized. They were obtained by condensation of piperonal (3,4-methylenedioxybenzaldehyde) with the corresponding aromatic primary amines. Their ¹H and 13C-NMR spectra have been obtained and the Hammett correlations including chemical shifts and the subsitutent constants (sigmap, sigmaR e sigmaI) were studied. Linear and bilinear significant correlations were observed for iminic carbon (C-alpha) and C-1’, showing a more significant ressonance effect on chemical shifts. The chemical shifts for C-4’ were highly affected by substituent effects, especially for halogens in the expected direction. Their biological activity against microorganisms has also been measured and significant activity was showed against Epidermophyton floccosum. The biological activity did not give a reasonable relationship with electronic effects.

Lipases have been immobilized in microemulsion-based organogels (MBG's) and successfully utilized for the enantioselective esterification, diesterification and transesterification reactions, in organic solvents at 25ºC. This methodology is described as a new alternative for the use of enzymes in organic solvents. High enzymic stability has been observed. We have also used this methodology for the successful resolution of chiral secondary alcohols. This is a convenient way of using this catalyst in organic solvents which employs small amounts of the enzyme (250mg/mL).

Amostras de água destilada, autoclavadas, foram inoculadas com L. monocytogenes cepa V7 e cepa VPH-1, e incubadas, aerobicamente, a 30ºC por 48 horas. Cada cepa foi testada individualmente, e determinou-se curvas de crescimento a 1, 2, 3, 4, 5, 21, 24, e 48 horas. O crescimento ou sobrevivência das duas cepas foi semelhante e encontrou-se sobreviventes em 24 horas de incubação. Examinou-se a atividade bactericida de um dos peptídeos catiônicos sintéticos (NP-2) contra L. monocytogenes cepa V7, em sistema aquoso. A atividade antibacteriana de NP-2 (1, 5, and 10µg/ml) foi melhor aos 60 minutos de incubação, com 10µg/ml de peptídeo, a 30 C.

Autoclaved distilled water samples were inoculated with L. monocytogenes strain V7 and strain VPH-1, and incubated aerobically, at 30 C for 48 hours. Each strain was tested individually, and growth curves were determined at 1, 2, 3, 4, 5, 21, 24, and 48 hours. The growth or survival of L. monocytogenes was similar for both strains, with survivors at 24 hour-incubation. The microbicidal activity of one synthetic cationic peptide (NP-2) was examined against L. monocytogenes strain V7, in a water system. Antibacterial activity of NP-2 (1, 5, and 10 g/ml) was best expressed at 60 minute-incubation, with 10 g/ml of peptide, at 30 C.

Amostras de água destilada, autoclavadas, foram inoculadas com L. monocytogenes cepa V7 e cepa VPH-1, e incubadas, aerobicamente, a 30ºC por 48 horas. Cada cepa foi testada individualmente, e determinou-se curvas de crescimento a 1, 2, 3, 4, 5, 21, 24, e 48 horas. O crescimento ou sobrevivência das duas cepas foi semelhante e encontrou-se sobreviventes em 24 horas de incubação. Examinou-se a atividade bactericida de um dos peptídeos catiônicos sintéticos (NP-2) contra L. monocytogenes cepa V7, em sistema aquoso. A atividade antibacteriana de NP-2 (1, 5, and 10µg/ml) foi melhor aos 60 minutos de incubação, com 10µg/ml de peptídeo, a 30 C.

Autoclaved distilled water samples were inoculated with L. monocytogenes strain V7 and strain VPH-1, and incubated aerobically, at 30 C for 48 hours. Each strain was tested individually, and growth curves were determined at 1, 2, 3, 4, 5, 21, 24, and 48 hours. The growth or survival of L. monocytogenes was similar for both strains, with survivors at 24 hour-incubation. The microbicidal activity of one synthetic cationic peptide (NP-2) was examined against L. monocytogenes strain V7, in a water system. Antibacterial activity of NP-2 (1, 5, and 10 g/ml) was best expressed at 60 minute-incubation, with 10 g/ml of peptide, at 30 C.