S514 Special Abstracts / Journal of Biotechnology 150S (2010) S1–S576

of the plant wall cell, as cellulose, hemicellulose, lignin and pectinsare degraded by enzymatic systems produced by a wide varietyof saprophytic and phythopathogenic microorganisms, includingbacteria and fungi. The aim of this work was to produce avicelase,carboxymethylcellulase, xylanase and pectinase from the exo-1mutant Neurospora crassa, FGSC 2256 and the wild type Neurosporacrassa, FGSC 424. The exo-1 mutant has a pleiotropic mutationwhich confers derepression of alpha-amylase, glucoamylase, beta-fructofuranosidase, and trehalase, as previously documented. Thismutation appears to also affect the composition of the cell wall,since segregants resulting from the backcross of exo-1 to thewild-type strain from which it derived are altered in the ratio ofgalactosamine to glucosamine in the cell wall. Conidial cell wallsof exo-1 mutant exhibit a marked decrease in the amount of galac-tosamine. In this work, increased levels (approximately sixfold) ofpectinase were found in conidia of exo-1 as compared with those ofthe wild type and also the polysaccharide-degrading activities werepreferentially induced and secreted by the exo-1 mutant comparedwith wild type cells, in the presence of inducers. Then, in culturemedia supplemented with 1% avicel from the exo-1 mutant wereobserved higher levels of avicelase (10.6-fold), carboxymethylcel-lulase (8.92-fold) and xylanase (4.74-fold) than in the wild type.Galactose was better inducer of pectinase than pectin and the exo-1 mutant produced three-fold more polygalacturonase than thewild type. Constitutive levels of polygalacturonase and cellulolyticenzymes were detected in medium devoid of carbon source. Thisresults show the importance of composition of the fungal cell wallin the regulation and secretion of industrial enzymes with applica-tion in the biomass treatment and biofuel biotechnology.

Support: FAPESP and CNPq

doi:10.1016/j.jbiotec.2010.09.815

[P-S.9]

Potential Application in Animal Feed of Phytase Produced fromAgro-Industrial Residues by Aspergillus japonicus

A. Maller 1, A.R.L. Damásio 1, T.M. Silva 2, J.A. Jorge 2, H.F. Terenzi 2,M.L.T.M. Polizeli 2,∗

1 São Paulo University, Departamento Bioquímica e Imunologia,FMRP/USP, Brazil2 São Paulo University, Departamento Biologia, FFCLRP/USP, BrazilKeywords: Fungal phytase; Assay enzymatic method; Aspergillus

Phosphorus is a important and limiting nutrient in biologicalsystems and phytic acid is the main form of storage of this com-post in vegetables. Phytases are phosphatases that hydrolyse theester bond liberating inositol and inorganic phosphate and theycan be divided in histidine acid, �-propeller and purple acid phos-phatase. Phytases have a great potential for biotechnological use inanimal feed and are mainly obtained from filamentous fungi. Theaim of this work was to produce phytase from A. japonicus undersolid state fermentation (SSF) using 11 different carbon sources;to standardize physico-chemical parameters for activity assay andverifying the enzymatic efficiency of inorganic phosphorous pro-duction from animal feed. The organism belongs to a collection ofour laboratory. It was inoculated in 4 g of several agro-industrialresidues plus 2 mL of distilled water, at 30 ◦C for 5 days. The assayswere carried out with 1% phytic acid dodecasodium in 100 mMsodium acetate buffer, pH 6.0, but other buffers and pH values werealso tested. The inorganic phosphorous formed was quantified withacidified ammonium molybdate using a modification of Yin et al.method (2007). Reducing sugar was quantified by Miller (1959). Totest the activity in feed animal (FATEC®), 1 g of this compost was

treated with 5 mL of phytase plus 5 mL of 100 mM sodium acetatebuffer, pH 6.0. Phytase activity released 632 �M.mL−1 of inorganicphosphorous and 259 �M.mL−1 of reducing sugar after 60 min ofincubation. Wheat bran was the best agro-industrial substrate forphytase production. It was observed a maximum activity in therange 30–50 ◦C and pH 3.5, but at pH 6.0 and 7.5 also was observedhigh activity, suggesting isoforms. Phytase was thermostable at50 ◦C, with a t50 of 40 min. This work showed that agro-industrialresidues are efficient inducers of phytase. The phytase producedby A. japonicus has potential application, improving the amountof free inorganic phosphate in the animal feed, resulting in gain ofweight, and diminishing the necessity of addition of this compoundto animal feed.

Support: FAPESP, CNPq

doi:10.1016/j.jbiotec.2010.09.816

[P-S.10]

Bioconversion of Bio-oils to hydrocarbon in Escherichia coli

Y.J. Choi 1,∗, J.H. Park 1, S.Y. Lee 1,2

1 Metabolic and Biomolecular Engineering National Research Labora-tory, Department of Chemical and Biomolecular Engineering (BK21Program) and BioProcess Engineering Research Center, KAIST, Daejeon,Republic of Korea2 Department of Bio and Brain Engineering, and BioinformaticsResearch Center, KAIST, Daejeon, Republic of KoreaKeywords: Hydrocarbon; Oils; Escherichia coli

Hydrocarbon is an organic compound consisting entirely ofhydrogen and carbon and the major components of natural gasand petroleum. Currently, bio-production of hydrocarbon has greatinterest as bio-gasoline has lots of interesting and outstandingbeneficial properties compared to petroleum-based gasoline. Mostdifferent thing compared to petroleum-based gasoline is that notonly the use of bio-gasoline maintains a balanced carbon diox-ide cycle due to based on renewable biological materials but alsoreduced carbon monoxide emissions during combustion. Further-more, it has high flash point and pure bio-gasoline or mixed withpetroleum-based gasoline can be used in conventional engineswithout any modifications. In spite of these positive ecologicalaspects, there is no report about production bio-gasoline. Here wereport the present study that assembly of novel synthetic pathwaysbased on known metabolic and regulatory information allowed forthe development of engineered Escherichia coli capable of produc-ing bio-gasoline using bio-oils. [This work was supported by theKorean Systems Biology Research Project (20090065571) of theMinistry of Education, Science and Technology (MEST) through theNational Research Foundation of Korea (NRF). Further supports bythe World Class University Program (R32-2008-000-10142-0) ofthe MEST, LG Chem Chair Professorship, IBM SUR program, andMicrosoft are appreciated.]

doi:10.1016/j.jbiotec.2010.09.817