17
views
0
recommends
+1 Recommend
1 collections
    0
    shares
      • Record: found
      • Abstract: found
      • Article: found
      Is Open Access

      PURIFICACIÓN Y CARACTERIZACIÓN DE UNA a-AMILASA PRODUCIDA POR LA CEPA NATIVA Bacillus sp. BBM1 Translated title: PURIFICATION AND CHARACTERIZATION OF A a-AMYLASE PRODUCED BY Bacillus sp. BBM1

      research-article

      Read this article at

      Bookmark
          There is no author summary for this article yet. Authors can add summaries to their articles on ScienceOpen to make them more accessible to a non-specialist audience.

          Abstract

          La cepa Bacillus sp. BBM1, productora de a-amilasas, fue aislada a partir de una muestra de suelo de la Universidad Nacional de Colombia sede Medellín. La caracterización morfológica, bioquímica y molecular indica que esta bacteria está filogenéticamente relacionada con las especies B. subtilis o B. amyloliquefaciens. La amilasa producida (BBM1) fue purificada por precipitación con sulfato de amonio y su peso molecular fue estimado en 77.6 kDa por electroforésis SDS-PAGE. Esta enzima es completamente estable a 60°C y presenta actividad significativa entre 30 y 80°C. La amilasa BBM1 tiene un pH óptimo entre 5.0-7.0 y no requiere calcio para su funcionamiento; estas propiedades hacen que esta enzima sea atractiva en aplicaciones industriales.

          Translated abstract

          A novel a-amylase producing strain, Bacillus sp. BBM1, was isolated a soil sample from Universidad Nacional de Colombia sede Medellín. Morphological, Biochemical and molecular data suggests that this strain belongs to either B. subtillis or B. amyloliquefaciens species. a-amylase BBM1 was purified by ammonium sulphate precipitation and its molecular weight estimated as 77.7 kDa by SDS-PAGE electrophoresis. This enzyme was completely stable at 60°C as presents significant activity in the 30-80°C range. Amylase BBM1 has an optimal pH in the range 5.0-7.0 and does not require calcium. All this properties make this enzyme interesting for future industrial applications.

          Related collections

          Most cited references29

          • Record: found
          • Abstract: found
          • Article: found
          Is Open Access

          Potential and utilization of thermophiles and thermostable enzymes in biorefining

          In today's world, there is an increasing trend towards the use of renewable, cheap and readily available biomass in the production of a wide variety of fine and bulk chemicals in different biorefineries. Biorefineries utilize the activities of microbial cells and their enzymes to convert biomass into target products. Many of these processes require enzymes which are operationally stable at high temperature thus allowing e.g. easy mixing, better substrate solubility, high mass transfer rate, and lowered risk of contamination. Thermophiles have often been proposed as sources of industrially relevant thermostable enzymes. Here we discuss existing and potential applications of thermophiles and thermostable enzymes with focus on conversion of carbohydrate containing raw materials. Their importance in biorefineries is explained using examples of lignocellulose and starch conversions to desired products. Strategies that enhance thermostablity of enzymes both in vivo and in vitro are also assessed. Moreover, this review deals with efforts made on developing vectors for expressing recombinant enzymes in thermophilic hosts.
            Bookmark
            • Record: found
            • Abstract: found
            • Article: not found

            Cloning, sequencing, characterization, and expression of an extracellular alpha-amylase from the hyperthermophilic archaeon Pyrococcus furiosus in Escherichia coli and Bacillus subtilis.

            A gene encoding a highly thermostable extracellular alpha-amylase from the hyperthermophilic archaeon Pyrococcus furiosus was identified. The gene was cloned, sequenced, and expressed in Escherichia coli and Bacillus subtilis. The gene is 1383 base pairs long and encodes a protein of 461 amino acids. The open reading frame of the gene was verified by microsequencing of the recombinant purified enzyme. The deduced amino acid sequence is 25 amino acids longer at the N terminus than that determined by sequencing of the purified protein, suggesting that a leader sequence is removed during transport of the enzyme across the membrane. The recombinant alpha-amylase was biochemically characterized and shows an activity optimum at pH 4.5, whereas the optimun temperature for enzymatic activity is close to 100 degrees C. alpha-Amylase shows sequence homology to the other known alpha-amylases and belongs to family 13 of glycosyl hydrolases. This extracellular alpha-amylase is not homologous to the subcellular alpha-amylase previously isolated from the same organism.
              Bookmark
              • Record: found
              • Abstract: found
              • Article: not found

              Hyperthermostable mutants of Bacillus licheniformis alpha-amylase: multiple amino acid replacements and molecular modelling.

              We have identified previously two critical positions for the thermostability of the highly thermostable alpha-amylase from Bacillus licheniformis. We have now introduced all 19 possible amino acid residues to these two positions, His133 and Ala209. The most favourable substitutions were to Ile and Val, respectively, which both increased the half-life of the enzyme at 80 degrees C by a factor of approximately 3. At both positions a stabilizing effect of hydrophobic residues was observed, although only in the case of position 133 could a clear correlation be drawn between the hydrophobicity of the inserted amino acid and the gain in protein stability. The construction of double mutants showed a cumulative effect of the most favourable and/or deleterious substitutions. Computer modelling was used to generate a 3-D structure of the wild-type protein and to model substitutions at position 209, which lies in the conserved (alpha/beta)8 barrel domain of alpha-amylase; Ala209 would be located at the beginning of the third helix of the barrel, in the bottom of a small cavity facing the fourth helix. The model suggests that replacement by, for example, a valine could fill this cavity and therefore increase intra- and interhelical compactness and hydrophobic interactions.
                Bookmark

                Author and article information

                Contributors
                Role: ND
                Role: ND
                Role: ND
                Journal
                dyna
                DYNA
                Dyna rev.fac.nac.minas
                Universidad Nacional de Colombia (Medellín )
                0012-7353
                June 2010
                : 77
                : 162
                : 31-38
                Affiliations
                [1 ] Universidad Nacional de Colombia Colombia
                [2 ] Universidad Nacional de Colombia Colombia
                [3 ] Universidad Nacional de Colombia Colombia
                Article
                S0012-73532010000200004
                94834118-d18a-4d84-867e-2662c9fe5b46

                http://creativecommons.org/licenses/by/4.0/

                History
                Product

                SciELO Colombia

                Self URI (journal page): http://www.scielo.org.co/scielo.php?script=sci_serial&pid=0012-7353&lng=en
                Categories
                ENGINEERING, MULTIDISCIPLINARY

                General engineering
                Bacillus,enzymes,a-amylase,enzimas,a-amilasa
                General engineering
                Bacillus, enzymes, a-amylase, enzimas, a-amilasa

                Comments

                Comment on this article