Purification, characterization and application of a novel extracellular agarase from a marine Bacillus megaterium

Khambhaty, Yasmin; Mody, Kalpana; Jha, Bhavanath

doi:10.1007/s12257-008-0026-3

Cited by 21 publications

(14 citation statements)

References 29 publications

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“…Strains of B. megaterium have been recognised for biological control of root infections due to Rhizoctonia solani [8]. Strains producing biomolecules, such as phytohormone, 2-pentylfuran, pyruvate, oxetanocin, vitamin B12, and polyhydroxyalkanoate (PHA) which are of industrial importance have been investigated [9][10][11][12]. This bacterium produces proteins of economical importance, namely, agarase, chitosanase, penicillin G acylase, amylases, glucose dehydrogenase, neutral protease, and the family of P-450 cytochrome monooxygenases [13][14][15].…”

Section: Introductionmentioning

confidence: 99%

Rapid Identification of Polyhydroxyalkanoate Accumulating Members of Bacillales Using Internal Primers forphaCGene ofBacillus megaterium

et al. 2013

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Bacillus megaterium is gaining recognition as an experimental model and biotechnologically important microorganism. Recently, descriptions of new strains of B. megaterium and closely related species isolated from diverse habitats have increased. Therefore, its identification requires several tests in combination which is usually time consuming and difficult to do. We propose using the uniqueness of the polyhydroxyalkanoate synthase C gene of B. megaterium in designing primers that amplify the 0.9 kb region of the phaC for its identification. The PCR method was optimized to amplify 0.9 kb region of phaC gene. After optimization of the PCR reaction, two methods were investigated in detail. Method I gave an amplification of a single band of 0.9 kb only in B. megaterium and was demonstrated by several strains of B. megaterium isolated from different habitats. The use of Method I did not result in the amplification of the phaC gene with other members of Bacillales. The specificity for identification of B. megaterium was confirmed using sequencing of amplicon and RT-PCR. Method II showed multiple banding patterns of nonspecific amplicons among polyhydroxyalkanoate accumulating members of Bacillales unique to the respective species. These methods are rapid and specific for the identification of polyhydroxyalkanoate accumulating B. megaterium and members of Bacillales.

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Section: Introductionmentioning

confidence: 99%

Rapid Identification of Polyhydroxyalkanoate Accumulating Members of Bacillales Using Internal Primers forphaCGene ofBacillus megaterium

et al. 2013

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“…The pH of the culture strongly affects enzymatic processes and transport of compounds across the cell membrane. Several carrageenan degrading bacteria Pseudomonas elongata [24], Cytophaga sp. [25], Cytophaga like bacterium [26][27][28] were found to produce carrageenases at pH range of 7-8 and few at different temperatures, ranging from 25 °C [15,29] to 37 °C [24,27] are reported.…”

Section: Discussionmentioning

confidence: 99%

“…It is commonly assumed that marine bacteria live in the sea, most of them are salt tolerant. Most carrageenan utilizing bacteria like P. elongata [24], P. alteromonas-like bacterium WZUC10 International Letters of Natural Sciences Vol. 17 203 [30], P. carrageenovora [7,31], Vibrio sp.…”

Section: Discussionmentioning

confidence: 99%

Increased Production of Carrageenase by <i>Pseudomonas aeruginosa</i> ZSL-2 Using Taguchi Experimental Design

Ziayoddin

Lalitha

Shinde

2014

ILNS

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The culture conditions for the production of carrageenase were optimized using one-factor-at-atime method combined with orthogonal array design. With the one-factor-at-a-time method revealed optimal conditions for carrageenase production were 24 h of fermentation period, 28 °C incubation temperature at pH 8.0 with NaNO 3 as nitrogen source and carrageenan as carbon source in MMS media. Further optimization of carrgeenase production by using orthogonal experimental design L 9 (3 4 ) with four factors, temperature, pH, NH 4 NO 3 and carrageenan with their relevant levels revealed optimised conditions for carrageenase production were temperature of 28 °C, pH 8.0, 2 g L -1 NaNO 3 and 2 g L -1 carrageenan. The order of the factors affecting the fermentation process was found to be temperature > pH > NaNO 3 > carrageenan. The temperature played a significant role on the carrageenase production. Higher carrageenase yield with activity of 0.542 ±0.045 U ml -1 was obtained in the optimised medium when compared to those of basal medium. Carrageenase hydrolysed products of carrageenan were identified by LC-ESI-MS as neocarrabiose, neocarrabiose-4 sulfate, neocarratetraose, neocarratetraose-4 sulfate, anhydrogalactose, galactose, galactose-4 sulphate and sulphate.

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“…3a). Esta capacidad de degradar agarosa a azúcares más simples ha sido previamente descrita para diversas especies del género Bacillus (e.g., Bacillus megaterium, Bacillus cereus y Bacillus subtilis) aisladas desde sedimento o algas rojas de zonas tropicales (Kim et al, 1999;Khambhaty et al, 2008;Saraswathi FIGURA 2: Árbol fi logenético de secuencias del gen ribosomal 16S inferido por los métodos de máxima verosimilitud y distancia (usando 1396 posiciones de alineamiento). Los números presentados corresponden a los valores de soporte (valores de bootstrap) de PhyML y Phylip NJ respectivamente.…”

Section: Discussionunclassified

Cepa antártica de Bacillus sp., con actividad extracelular de tipo agarolítica y alginatoliasa

et al. 2013

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RESUMENDiversas bacterias asociadas a macroalgas pueden usar fi cocoloides como fuente de carbono. Las bacterias de la Antártica, poseen características fi siológicas adaptativas que habrían evolucionado para permitir su supervivencia y funcionamiento dentro de las condiciones adversas de ese ecosistema. Por lo tanto, bacterias aisladas desde algas antárticas podrían tener la capacidad de degradar enzimáticamente azúcares complejos a temperaturas inferiores que aquellas aisladas desde zonas más cálidas, lo que potencialmente podría tener aplicaciones en la mejora en procesos industriales que utilizan enzimas. El análisis del gen ribosomal 16S indica que la cepa bacteriana aislada desde algas varadas en la Isla Rey Jorge (Antártica), pertenece al género Bacillus. El sobrenadante libre de células presentó actividad agarasa y alginatoliasa. Se encontraron diferencias signifi cativas en la temperatura óptima para hidrolizar agarosa y alginato, evaluada dentro de un rango de 4 a 30 °C. La actividad agarolítica fue mayor a 4 °C, mientras que la actividad alginatoliasa fue mayor a 30 °C. Estos resultados poseen un alto valor biotecnológico y podría ser utilizada con fi nes industriales. PALABRAS CLAVE:Bacillus, Antártica, Agarasa, Alginatoliasa. ABSTRACTSeveral bacteria associated to macroalgae can use phycocolloids as carbon source. Antarctic Bacteria's have physiological characteristics that might have evolved to allow the survival and functioning under the harsh conditions of that ecosystem. Therefore, Antarctic bacteria isolated from algae should have the ability to degrade complex sugars at lower temperatures than those isolated from warmer areas, which may have applications in the improvement of industrial processes that uses enzymes. The bacterial strain isolated from wrack algae, in King George Island, Antarctica, was identifi ed as Bacillus on the base of 16S ribosomal gene analysis. The cell-free supernatant of the culture medium showed alginate-lyase and agarase activities. Signifi cant differences in the optimal temperature to hydrolyze agarose and alginate were found within a range of 4 to 30 °C. While the agarase activity was higher at 4° C, the alginate-lyase activity was higher at 30 °C. Our results have biotechnological value and could be used with industrial aims.

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Purification, characterization and application of a novel extracellular agarase from a marine Bacillus megaterium

Cited by 21 publications

References 29 publications

Rapid Identification of Polyhydroxyalkanoate Accumulating Members of Bacillales Using Internal Primers forphaCGene ofBacillus megaterium

Rapid Identification of Polyhydroxyalkanoate Accumulating Members of Bacillales Using Internal Primers forphaCGene ofBacillus megaterium

Increased Production of Carrageenase by <i>Pseudomonas aeruginosa</i> ZSL-2 Using Taguchi Experimental Design

Cepa antártica de Bacillus sp., con actividad extracelular de tipo agarolítica y alginatoliasa

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