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Ítem Biodegradation of vegetable residues by polygalacturonase-agar using a trickle-bed bioreactor(Institution of Chemical Engineers, 2018-09-01) Ramírez-Tapias, Y.A.; Rivero, C.W.; Giraldo-Estrada, C.; Britos, C.N.; Trelles, J.A.; Universidad EAFIT. Departamento de Ciencias; Ciencias Biológicas y Bioprocesos (CIBIOP)Bacterial pectinases degrade the pectic substances present in plant tissues and particularly, polygalacturonases catalyze the hydrolysis of a-(1,4) glycosidic bonds linking D-galacturonic acid units. In this study, polygalacturonase from Streptomyces halstedii ATCC 10897 was immobilized by the matrix entrapment technique using different thermogels. Bacteriological agar added with magnesium cation produced beads with a more stabilized microstructure for enzyme retention, monitored by oscillatory measurements of storage and loss modulus. Agar concentration and protein content were optimized to maximize protein entrapment, product conversion, and reaction yield. Results showed that the mixture at 10:90% (v/v) of protein (2 mg/mL) and agar (4% w/v) was the best immobilization condition to retain 91% of protein and hydrolyze 38% of pectin to allow the highest reaction yield (9.279 g/g) and increase stability up to 48 h of successive reactions. Agarose bead biocatalysts were used in a trickle-bed column operated with recirculation, and this bioreactor allowed the degradation of pear and cucumber residues by enzymatic liquefaction to enhance sugar content up to 15.33 and 9.35 mg/mL, respectively, and decrease viscosity by 92.3%. The scale-up of this process adds value to vegetable residues such as fructooligosaccharides or fermentable sugars, which become a sustainable source of fuels and chemicals. © 2018 Institution of Chemical EngineersÍtem Enhanced molecular visualization of root colonization and growth promotion by Bacillus subtilis EA-CB0575 in different growth systems(Elsevier GmbH, 2018-01-01) Posada, L.F.; Álvarez, J.C.; Romero-Tabarez, M.; de-Bashan, L.; Villegas-Escobar, V.; Universidad EAFIT. Departamento de Ciencias; Ciencias Biológicas y Bioprocesos (CIBIOP)Bacillus subtilis EA-CB0575 is a plant growth-promoting bacterium (PGPB) associated with banana and tomato crops. Root colonization is an important trait for PGPB microorganisms and potentiates the bacterial effect related to the mechanisms of plant growth promotion. Therefore, detection of bacterial colonization of roots in different culture systems is important in the study of plant–microorganism interactions. In this study, fluorescent in situ hybridization (FISH) and catalyzed reporter deposition–FISH (CARD–FISH) were evaluated to determine the colonization ability of B. subtilis EA-CB0575 on banana and tomato roots planted on solid and liquid Murashige and Skoog medium (MS(S) and MS(L), respectively) and in soil for tomato plants. Results showed B. subtilis colonization 0–30 days post inoculation for banana and tomato plants in different culture systems with differential distribution of bacterial cells along tomato and banana roots. FISH and CARD–FISH methodologies were both successful in detecting B. subtilis colonies, but CARD–FISH proved to be superior due to its enhanced fluorescence signal. The presence of bacteria correlated with the promotion of plant growth in both plant species, providing clues to relate rhizospheric colonization with improvement in plant growth. FISH and CARD–FISH analysis results suggested the presence of native microbiota on the roots of in vitro banana plants, but not on those of tomato plants. © 2018 Elsevier GmbHÍtem Enhanced molecular visualization of root colonization and growth promotion by Bacillus subtilis EA-CB0575 in different growth systems(Elsevier GmbH, 2018-01-01) Posada, L.F.; Álvarez, J.C.; Romero-Tabarez, M.; de-Bashan, L.; Villegas-Escobar, V.; Universidad EAFIT. Departamento de Ciencias; Biodiversidad, Evolución y ConservaciónBacillus subtilis EA-CB0575 is a plant growth-promoting bacterium (PGPB) associated with banana and tomato crops. Root colonization is an important trait for PGPB microorganisms and potentiates the bacterial effect related to the mechanisms of plant growth promotion. Therefore, detection of bacterial colonization of roots in different culture systems is important in the study of plant–microorganism interactions. In this study, fluorescent in situ hybridization (FISH) and catalyzed reporter deposition–FISH (CARD–FISH) were evaluated to determine the colonization ability of B. subtilis EA-CB0575 on banana and tomato roots planted on solid and liquid Murashige and Skoog medium (MS(S) and MS(L), respectively) and in soil for tomato plants. Results showed B. subtilis colonization 0–30 days post inoculation for banana and tomato plants in different culture systems with differential distribution of bacterial cells along tomato and banana roots. FISH and CARD–FISH methodologies were both successful in detecting B. subtilis colonies, but CARD–FISH proved to be superior due to its enhanced fluorescence signal. The presence of bacteria correlated with the promotion of plant growth in both plant species, providing clues to relate rhizospheric colonization with improvement in plant growth. FISH and CARD–FISH analysis results suggested the presence of native microbiota on the roots of in vitro banana plants, but not on those of tomato plants. © 2018 Elsevier GmbH