fevereiro 2015 vol. 1 num. 2 - XX Congresso Brasileiro de Engenharia Química
Artigo - Open Access.
Production of Recombinant Enzymatic Mixture by Co-culture Strategy
Sugarcane bagasse is considered to be a residue with great potential as feedstock for second-generation ethanol production, and other added-value products. However, certain obstacles must be overcome to make this technology viable, as in the case of enzymatic route in which the high cost of enzymes production, and the low efficiency in the saccharification step still represent a bottleneck. Recently, recombinant enzymes have been used as supplement fungal cocktails to improve lignocellulosic biomass saccharification, as well as to assembly specific enzymatic mixtures aiming at other applications. In this study, we present the development of recombinant enzymatic mixture production process based on co-culture strategy. The process development includes the inoculum assembly, screening of alternative carbon sources, and improve of induction phase. Escherichia coli clones co-culture present as main advantages: the possibility to produce an enzymatic mixture with a desired composition based on inoculum assembling, the use of low cost carbon source as hemicellulosic hydrolysates based on complete metabolic machinery of E. coli, and reduction of the production process and recovery costs since more than one enzyme is produced in the same reactor.
-  BRADFORD, M.M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry, 72 (1-2) (1976) 248-54.
-  GAO, D.; UPPUGUNDLA, N.; CHUNDAWAT, P.S.; YU, X.; HERMANSON, S.; GOWDA, K.; BRUMM, P.; MEAD, D.; BALAN, V.; DALE, D.B. Hemicellulases and auxiliary enzymes for improved conversion of lignocellulosic biomass to monosaccharides. Biotechnology for Biofuels, (2011) 4:5 GOMES, D.S. Produção de misturas enzimáticas recombinantes utilizando a técnica de co-cultivo. Relatório de estágio realizado no Laboratório Nacional de Ciência e Tecnologia do Bioetanol – CTBE/ CNPEM, (2013) LEE, S.Y. High cell density culture of Escherichia coli. Tibtech, 14 (1996) 98–105.
-  Área temática: Processos Biotecnológicos 7SHIMIZU, K. Metabolic Regulation of a Bacterial Cell System with Emphasis on Escherichia coli Metabolism. Biochemistry, (2013) 47.
-  LAEMMLI, U.K. Cleavage of structural proteins during the assembly of the head of bacteriophage T Nature, 2273 (1970) 397–407.
-  MILLER, G.L. Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal. Chem, 31 (1959) 426–428.
-  NEHME, N.; MATHIEU, F.; TAILLANDIER, P. Impact of the co-culture of Saccharomyces cerevisiae–Oenococcus oeni on malolactic fermentation and partial characterization of a yeast-derived inhibitory peptidic fraction. Food Microbiology, 27 (2010) 150–157.
-  OLAEF, O.A.; BURTON, S.G.; COWAN, D.A.; HARRISON, S.T.L. Improving of a thermostable amidase through optimising IPTG induction in a highly dense culture of recombinant Escherichia coli. Biochemical Engeneering journal. 52 (2010) 19-24.
-  SHIN, H.D.; MCCLEDON, S.; VO, T.; CHEN, R.R. Escherichia coli binary culture engineered for direct fermentation of hemicellulose to a biofuel. Applied and environmental microbiology, 76 (24) (2010) 8150-8159 ZHOU, J.; WANG, Y.H.; CHU, J.; LUO, L.Z.; ZHUANG, Y.P.; ZHANG, S.L. Optimization of cellulase mixture for efficient hydrolysis of steam-exploded corn stover by statistically designed experiments. Bioresour Technol, 100 (2009) 819-825. PubMed doi:10.1016/j.biortech.20006.068
SILVA, M. R.; GOMES, D. S.; FREITAS, S.; "Production of Recombinant Enzymatic Mixture by Co-culture Strategy", p. 1193-1200 . In: Anais do XX Congresso Brasileiro de Engenharia Química - COBEQ 2014 [= Blucher Chemical Engineering Proceedings, v.1, n.2].
São Paulo: Blucher,
ISSN 2359-1757, DOI 10.5151/chemeng-cobeq2014-0799-23741-175193
últimos 30 dias | último ano | desde a publicação