Special Abstracts / Journal of Biotechnology 150S (2010) S1–S576 S419

database is open source and accessible at the following URL:www.ylichron.it/PHA pseudomonas DB.

doi:10.1016/j.jbiotec.2010.09.572

[P-I.207]

Investigation of Liquid and Solid Fermentation Processes of theFungus Aspergillus fumigatus for Protease Production

R.R. Silva 1, A. Rodrigues 2, H. Cabral 3,∗

1 Instituto de Biociências, Letras e Ciências Exatas - UNESP, Brazil2 Universidade Estadual de Santa Cruz, Brazil3 Faculdade de Ciências Farmacêuticas de Ribeirão Preto – USP, BrazilKeywords: Protease; Aspergillus fumigatus; Fermentation

The production of microbial enzymes obeys the proper con-ditions of each microorganism species and the means applied tofermentation, being subject to the differences between liquid andsolid fermentations. This way, filamentous fungi have been widelyexploited as a productive source of proteases due to their growthcapacity and a wide production range. The fungus A. fumigatus wasisolated from the soil having decaying wood. Production studieswere conducted through liquid-state fermentation in Erlenmeyerflasks of 250 mL with 50 mL containing KH2PO4, 0,7%; K2HPO4,0,2%; MgSO4.7H2O, 0,01%; citrate 2H2O, 0,05%; yeast extract, 0,1%;CaCl2.2H2O, 0,01% and peptone 1% with supplementation of caseinand glucose. The solid fermentation was performed in Erlenmeyerflasks of 250 mL using wheat bran as a substrate adding proteinsupplementation, e.g. ovalbumin. The partial characterization wasdone with partially-purified enzyme through ethanol precipitation.The partial characterization was done by using azocasein as a sub-strate. The peak protease production in liquid fermentation was of96 hours, the same production period obtained for the solid one.The protease resulted from liquid fermentation presented pH andoptimum temperature of 8,0 and 50 ◦C, respectively and thermosta-bility of 50% after a 60-minute incubation at 45 ◦C, and belongsto the class of the serine proteases. The protease obtained fromsolid fermentation presented pH and optimum temperature of8,0 and 50 ◦C, respectively and thermostability of 75% at 45 ◦C for60 minutes. We conclude that the proteases produced by the fungusA. fumigatus in liquid and solid fermentation processes present thesame biochemical characteristics, except for the thermostability.Future experiments of purification and biochemical characteriza-tion will be carried out for protease obtained by solid fermentation.Financial support: FAPESP N◦ 2008/11303-6.

doi:10.1016/j.jbiotec.2010.09.573

[P-I.208]

Production of xylanase, cellulase and laccase by mesophilicfungi isolated from sugar cane soil

Tassia Chiachio Egea ∗, Andre Rodrigues, Roberto da Silva, EleniGomes

UNESP, BrazilKeywords: Enzyme; Fungi; Isolation; Soil

Isolation and identification of microorganisms with ability toproduce biocatalysts is of great importance for presentation of“new” enzymes to the biotechnological business. Isolation is alsoused as a tool that provides a survey of microorganisms capa-ble of metabolizing potentially toxic chemical compounds and isessential in the bioremediation, acting as a mitigation measure to

the problems of irrational use of pesticides. In this study, wereisolated 75 fungi strains from sugar cane soil, with different car-bon sources (glucose, sugar-cane bagasse, wheat bran and filterpaper) and at different times of the crop season. Among the isolates,were found 10 different genus, such as Trichoderma (39%), Fusarium(16%) and Cunninghamela (14%). The genera Aspergillus, Cunning-hamela, Fusarium and Trichoderma were isolated in all carbonsources media. The genera Cunninghamela, Mucor, and Trichodermawere the constant in the samples and only Trichoderma and Fusar-ium were found in all sample collected. The isolates were usedfor a solid-state fermentation, with sugarcane bagasse and wheatbran as substrates, to evaluate their potential to produce xylanase,CMCase and laccase. The enzyme production was quite varied,with xylanase activity reaching levels of 59.9 U/mL (Aspergillus sectfumigate) and 64,0 U/mL (Aspergillus sect flavus). The productionof CMCase was 4,7 U/mL for Trichoderma sp. and 4.2 U/mL forAspergillus sect nigri and laccase was determined as 0.5 U/mL forVerticilium sp. and 0.4 U/mL for Fusarium sp..

doi:10.1016/j.jbiotec.2010.09.574

[P-I.209]

Selection of the best source of carbon for production of recom-binants enzymes in liquid fermentation

Bárbara Garcia São Jóse, Fabiana Carina Pavezzi ∗, Cèlia Maria LandiFrando, Eleni Gomes, Roberto da Silva

UNESP, BrazilKeywords: Enzyme; Glucoamylase; Production; Starch

Glucoamylase (GA) is of great importance to the fermentationand food industries for saccharification of starch and other relatedpoly and oligosaccharides. The principal industrial use of GA isin the production of glucose and fructose syrups, which in turnserve as a feedstock for biological fermentations in the productionof ethanol. The enzyme is produced by a variety of microorgan-isms, in which Aspergillus niger, Aspergillus awamori and Rhizopusoryzae have been considered as the most important for indus-trial application. It is also produced by transformed Saccharomycescerevisiae, strains SRE15 and TH58. The purpose of this study wasto produce two thermostable recombinants GAs SRE15 and TH58using different culture medium in liquid fermentation. Two percentof starch from different sources was used soluble starch, cassavastarch, potato starch, corn starch, wheat starch; also brans of cas-sava and wheat were used as carbon source. The starches and branswere gelatinized at 100 ◦C for 3 min and, afterwards, the follow-ing items were added in each solution: 0.5% ammonium sulphate,2% glucose, 0.17% “Yeast nitrogen base without aminoacids”, L-histidin 20 mg/L, thus constituting seven diverse culture mediums.The strains SRE15 produced GA with high specific activity, 41.7,25.9 and 25.6U/mgprotein for the substrates: cassava bran, potatostarch and soluble starch, respectively. But lower specific activ-ity 4.0U/mgprotein for the substrate corn starch. The strains TH58also show high specific activity 30.0, 26.3 and 23.0U/mg proteinfor the substrates: soluble starch, cassava bran, and cassava starch.The cassava bran and soluble starch were best substrate for GAproduction by both strains.

Financial support: FAPESP

doi:10.1016/j.jbiotec.2010.09.575