title: Extracellular endoglucanase activity from Paenibacillus polymyxa BEb-40: production, optimization and enzymatic characterization.
abstract: Endoglucanase activity produced by Paenibacillus polymyxa BEb-40 was studied. In submerged culture with minimal medium supplemented with carboxymethylcellulose (CMC), this microorganism produced up to 0.37 U/mL endoglucanase activity with high specific activity (14.3 U/mgtotal protein). Detection of endoglucanase activity through zymography revealed at least 14 isoenzymes with molecular weights between 38 and 220 kDa. This high variety of secreted endoglucanases has not been described previously in Paenibacillus genus. The optimum conditions, determined by response surface methodology, were 48 °C and pH 3.4, which allowed an increase of 33.7 % in the relative endoglucanase activity obtained with respect to the standard conditions. Nevertheless, high levels of hydrolysis of at least 70 % of the maximum activity could be obtained at wide ranges of pH (2-9) and temperature (40-60 °C). Under optimal conditions, high levels of CMC hydrolysis were reached, of about 40 %, after only 12 h of reaction with substrate/total protein ratios between 19 and 76. Kinetic analysis revealed that endoglucanase activity followed a mixed inhibition model (K m = 8.4 mM, K ic = 0.03 mM, K iu = 0.35 mM, V max = 33.3 U/mgtotal protein). These results allow to consider P. polymyxa BEb-40 as a promising microorganism for the production of endoglucanases, with possibilities of application in the breakdown of lignocellulosic biomass. The high specific activity at wide ranges of pH and temperature can allow its use in a wide variety of processes, under both acidic and alkaline conditions, as well as in mesophilic and thermophilic temperatures, further reducing the amount of enzymes used.
authors.affiliation: Departamento de Biotecnología y Bioquímica, Centro de Investigación y de Estudios Avanzados del IPN, Unidad Irapuato. km 9.6 Libramiento Norte, Carr. Irapuato-León, Apdo. Postal 629, 36821, Irapuato, Guanajuato, México.
authors.forename: Argel
authors.initials: A
authors.lastname: Gastelum-Arellanez
authors.forename: Octavio
authors.initials: O
authors.lastname: Paredes-López
authors.forename: Víctor
authors.initials: V
authors.lastname: Olalde-Portugal
mesh_terms: D002266:Carboxymethylcellulose Sodium / Q000378:metabolism*; D002480:Cellulase / Q000737:chemistry / Q000378:metabolism*; D003470:Culture Media / Q000737:chemistry; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D004275:DNA, Ribosomal / Q000737:chemistry / Q000235:genetics; D004795:Enzyme Stability; D006863:Hydrogen-Ion Concentration; D007527:Isoenzymes / Q000737:chemistry / Q000378:metabolism; D007700:Kinetics; D008969:Molecular Sequence Data; D008970:Molecular Weight; D056507:Paenibacillus / Q000145:classification / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D017422:Sequence Analysis, DNA; D013696:Temperature
chemical_list: D003470:Culture Media; D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D007527:Isoenzymes; D012336:RNA, Ribosomal, 16S; D002480:Cellulase; D002266:Carboxymethylcellulose Sodium