Abstract The total cost of processes that use lipases can be reduced by using the mycelium or not purified crude extract with lipolytic activity. These catalysts can be used without complex purification and can show desirable properties to emerging applications like hydrolysis and biotransformation of vegetable oils. To explore the potential of multiple forms of lipases (produced by the same filamentous fungus species), this work investigated intracellular and extracellular lipases produced by Trichoderma koningii (intracellular lipases were never studied), Penicillium spp. F02 (isolated from industrial grease trap) and Aspergillus niger. For this, batch fermentations were conducted for 96h using olive oil as a carbon source. The submerged culture was interrupted with vacuum filtration to isolate the mycelium and crude extract. The lipolytic activities were determined by the hydrolysis method of olive oil. The best results were obtained for intracellular activity with T. koningii (81.5 U/g of dry biomass) and for extracellular activity with A. niger (10.6 U/mL of crude extract). Both were attained in 96h of fermentation. T. koningii showed an accumulation of intracellular lipases and a depreciated extracellular activity in the initial 24 hours, which can be reasonable indications that there is strong retention of lipases in the mycelium. Statistical analysis within the range of 37°C to 44°C associated with pH from 7.8 to 8.0 favored the activity of lipases from T. koningii. All these findings are promising for future studies in the enzymatic synthesis of biodiesel with lipases (intracellular and extracellular) from T. koningii. oils species, species , species) studied, studied studied) spp F F0 isolated trap h source T 81.5 815 81 5 (81. Ug U g biomass A 10.6 106 10 6 (10. UmL mL . extract) fermentation 2 hours 37C C 37 44C 44 78 7 8 7. 80 0 8. 81. (81 10. 1 (10 3 4 (8 (1 (

RESUMO Uma variedade de estudos tem sido realizados com o intuito de produzir celulases de baixo custo para serem utilizadas na produção de álcool de segunda geração (2G) a partir de resíduos lignocelulósicos. Dentre eles destaca-se o uso de bagaço de cana-de-açúcar como fonte de carbono para o crescimento dos microrganismos. Baseado nisso, este artigo teve como objetivo realizar uma revisão bibliométrica sobre o uso do Aspergillus niger empregando como fonte de carbono o bagaço de cana-de-açúcar, pretendendo a produção de celulase, através do estudo dos avanços científicos e tecnológicos sobre os temas. Para efetuar a revisão, recorreu-se ao banco de dados Web of Science, inserindo-se quatro palavras-chave: “Aspergillus niger”, “cellulase”, “cane” e “submerged” entre os anos de 1945 e 2018. A partir desta pesquisa, foi possível apontar os autores, periódicos e grupos de pesquisas mais importantes neste assunto. Identificou-se um aumento do número de publicações ao longo dos anos por diversos países, revelando o interesse pela produção de celulases de menor custo para geração de álcool 2G. O Brasil foi o país com o maior número de publicações sobre o tema, devido a grande capacidade de plantio de cana-de-açúcar e produção de álcool. Pela análise das redes de co-citação dos autores verificou-se a existência de uma relação dos grupos de pesquisa no Brasil, entre as cidades de Campinas, São Paulo e Curitiba e outra associação na Dinamarca, entre as cidades de Lyngby e Copenhagen.

ABSTRACT A variety of studies have been realized with the objective of producing celulases with low cost to be used in the production of second generation alcohol (2G) from lignocellulosic residues. Among them, the use of sugarcane bagasse as carbon source for the growth of microorganisms is highlighted. Based on this, this article aimed to carry out a bibliometric review on the use of Aspergillus niger using sugarcane bagasse as a carbon source, aiming the production of cellulase, through the study of scientific and technological advances on the themes. To perform the review, we used the Web of Science database, inserting four keywords: "Aspergillus niger", "cellulase", "cane" and "submerged" between the years of 1945 and 2018. From this search, was possible to identify the most important authors, periodicals and research groups in this subject. It was identified an increase in the number of publications over the years by several countries, showing interest in the production of lower cost cellulases for 2G alcohol generation. Brazil was the country with the largest number of publications on the subject, due to the great capacity of planting sugarcane and production of alcohol. By analyzing the correlation networks of the authors it was verified the existence of a relation of the research groups in Brazil, between the cities of Campinas, São Paulo and Curitiba and another association in Denmark, between the cities of Lyngby and Copenhagen.

Oily waste from sanitation processes was characterized for its potential use in the generation of biofuels. The waste residues studied showed high levels of oil and grease, reaching up to 87% (m/m) in a grease trap of a food company, showing that these residues can be utilized for the production of biofuels. The results revealed high levels of moisture and saponification as well as high levels of free fatty acids (FFA). Analysis of chromatographic profiles (HPLC) showed great heterogeneity of the fatty acids contained in the sample, and a predominance of palmitic, oleic, linoleic and linolenic acids.

O objetivo deste trabalho foi caracterizar a enzima penicilina G acilase (PGA) produzida por Bacillus megaterium, uma importante enzima industrial que catalisa a hidrólise de penicilina G, para produção de antibióticos semi-sintéticos. Purificação da enzima por ultra/diafiltração não permitiu detectar a banda de PGA por eletroforese SDS-PAGE devido ao elevado conteúdo de outras proteínas remanescentes. Contudo, utilizando DNA do microrganismo que vem sendo estudado, foi possível amplificar os genes das duas sub-unidades de PGA previstas na literatura, mostrando que a enzima em estudo é também constituída de duas sub-unidades, 245 e 537 aminoácidos cada, com massas moleculares médias de 26950 e 59070 Da, respectivamente. Foram estudadas as influências da temperatura 25-60(0)C, pH 5-10, e concentração do substrato na velocidade da reação de hidrólise da penicilina G. A temperatura e pH ótimos foram de 37(0)C e 8,0, respectivamente. O modelo de Michaelis-Menten representou bem a cinética da reação, com valores de parâmetros estimados de 1,83 mM para Km e Vmáx= 0,165*10-3 mmol/min/UI.

The purpose of this work was to characterize the enzyme penicillin G acylase (PGA) produced by Bacillus megaterium. Purification of the enzyme by ultra/diafiltration did not allow the detection of the PGA band by SDS-PAGE electrophoresis due to the high content of remaining proteins. However, using the DNA of the microorganism, it was possible to replicate the genes of the two B. megaterium PGA reported in literature, showing that the enzyme consisted of two sub-units, having 245 and 537 amino acids each and an average molecular mass of 26950 and 59070 Da, respectively. The parameters studied were: 1) the influence of temperature in the 25-60(0)C range, 2) pH in the 5-10 range and 3) substrate concentration, this was tested to obtain results on the Penicillin G hydrolysis reaction rate, using the initial velocities approach. The maximum hydrolysis rate was obtained at 37ºC and pH 8.0. The Michaelis-Menten model fitted well, resulting in estimated Km and Vmax parameters values of 1.83 mM and 0.165*10-3 mmol/min/UI, respectively.