Purification and characterization of an extracellular alpha-amylase from Clostridium perfringens type A

Shih, Neng-Jen; Labbé, Ronald G.

doi:10.1128/aem.61.5.1776-1779.1995

Cited by 25 publications

(6 citation statements)

References 29 publications

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“…This result could classify the enzyme as α-amylase with ability for liquefaction and saccharification. Similar results were reported by Shih and Labbe (1995) which found that the degradation of starch by purified C. perfringens amylase yielded products with a high degree of starch hydrolysis but low levels of reducing sugars indicating an endohydrolysis pattern. As expected, exo-acting enzymes like Starch Regular sugar amylase and amyloglucosidase gave high amounts of reducing sugar but low degrees of starch hydrolysis.…”

Section: Characterization Of -Amylase A3supporting

confidence: 89%

“…This value was confirmed by SDS-PAGE, and appeared as single subunits ( Figure 5). Similar molecular weights were reported for α-amylases from Aspergillus chevalieri (68 kDa) (Olutiola and Nwaogwugwu, 1982) and perfringens (76 KDa) (Shih and Labbe, 1995).…”

Section: Purification Of α-Amylase From T Harzianumsupporting

confidence: 81%

“…Amylolytic enzymes are metalloenzymes with up to six Ca 2+ atoms at the active site whose catalytic activity can be affected by mono-and divalent metals (Normurodova et al, 2007). Ca 2+ enhanced the activity of α-amylases from C. perfringens (Shih and Labbe, 1995), B. subtilis (Asgher et al, 2007) and Acremonium sporosulcatum (Valaparla, 2010). T. harzianum α-amylase A3 was partially inhibited by Zn 2+ and Ni 2+ , while Co 2+ had strong inhibitory effect.…”

Section: Characterization Of -Amylase A3mentioning

confidence: 99%

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Production, purification and characterization of α-amylase from Trichoderma harzianum grown on mandarin peel

Mohamed¹,

Azhar²,

Baakdah³

et al. 2011

Afr. J. Microbiol. Res.

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The production, purification and characterization of α-amylase from Trichoderma harzianum grown on mandarin peel were investigated. The effect of incubation time and mandarin peel concentration on the production of α-amylase by T. harzianum was studied. α-Amylase A3 was purified from T. harzianum to electrophoretic homogeneity by using DEAE-Sepharose and Sephacryl S-200 columns. The enzyme had molecular weight of 70 kDa using gel filtration and SDS-PAGE. The affinity of the substrates toward A3 was in the order of amylopectin > glycogen > starch > β-cyclodextrin > dextrin > α-cyclodextrin. These findings tend to suggest that the enzyme has high affinity toward high-molecular mass substrates. The K m and V max values of the enzyme for hydrolyzing potato soluble starch and glycogen were 6.53, 4.5 mg/ml and 2 and 2.2 μmol reducing sugar/ml, respectively. The maximum activity of enzyme against soluble starch was determined at pH 4.5 and 40°C. α-Amylase A3 was stable up to 40°C for 30 min of incubation and retained 70 and 50% of its activity at 50 and 60°C, respectively. While all the examined metal cations were effective in inhibiting the enzyme, Ca 2+ considerably enhanced the activity. The metal chelators, EDTA, sodium citrate and sodium oxalate had inhibitory effects on A3. The rate of breakdown of starch was higher than the rate of formation of reduced sugar indicating A3 is endoacting enzyme. These properties of A3 with its remarkable activity meet the prerequisites needed for liquefaction and saccharification of starch industry.

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Section: Characterization Of -Amylase A3supporting

confidence: 89%

Section: Purification Of α-Amylase From T Harzianumsupporting

confidence: 81%

Section: Characterization Of -Amylase A3mentioning

confidence: 99%

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Production, purification and characterization of α-amylase from Trichoderma harzianum grown on mandarin peel

Mohamed¹,

Azhar²,

Baakdah³

et al. 2011

Afr. J. Microbiol. Res.

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“…C. perfringens NCTC 8679 is able to sporulate in the presence of glucose (8 mM) in the presence of 30 mM organic acids (acetic, lactic, or butyric acid) (20). To determine the possible role of such acids as inducers of sporulation, their presence in the CSF of vegetative cultures of strain NCTC 8679 was examined by high-pressure liquid chromatography with an Aminex PHX-87H hydrogen column (Bio-Rad, Hercules, Calif.).…”

Section: The Culture Supernatant Fluids (Csfs) Of 12 Strains Of Clostmentioning

confidence: 99%

Sporulation-promoting ability of Clostridium perfringens culture fluids

Shih

Labbé

1996

Appl Environ Microbiol

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The culture supernatant fluids (CSFs) of 12 strains of Clostridium perfringens types A, B, C, and D stimulated sporulation of test strains NCTC 8238 and NCTC 8449 of this organism. The sporulation-promoting ability was present in vegetative and sporulating CSFs of both enterotoxin-positive (Ent+) and Ent- strains. The sporulation factor possessed a molecular weight between 1,000 and 5,000 and was heat and acid stable. This study suggests a potential role for Ent- strains in food-borne disease outbreaks caused by Ent+ strains of C. perfringens type A.

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“…These may be crucial for the overall hydrogen production rather than the original mixed cultures (Hiligsmann et al, 2011). Clostridia are known to grow on both polysaccharidic and proteinaceous substrates, and possess necessary hydrolytic enzymes, such as cellulases (Demain et al, 2005), xylanases (Thomas et al, 2014), amylases (Shih and Labbé, 1995), or proteases (Janoir et al, 2004). Nevertheless, some differences in the utilization of carbonaceous substrates among the used clostridial strains could be observed (Beckers et al, 2010).…”

Section: Introductionmentioning

confidence: 99%

Carbon source utilization and hydrogen production by isolated anaerobic bacteria

Jame

Zelená

Lakatoš

et al. 2016

Acta Chimica Slovaca

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Five bacterial isolates were tested for their ability to generate hydrogen during anaerobic fermentation with various carbon sources. One isolate from sheep rumen was identified as Escherichia coli and four isolates belonged to Clostridium spp. Glucose, arabinose, ribose, xylose, lactose and cellobiose were used as carbon sources. Results showed that all bacterial strains could utilize these compounds, although the utilization of pentoses diminished growth yield. The excretion of monocarboxylic acids (acetate, propionate, formiate, butyrate) into medium was changed after replacing glucose by other carbon sources. Di- and tricarboxylic acids were excreted in negligible amounts only. Spectra of excreted carboxylic acids were unique for each strain and all carbon sources. All isolates produced H2 between 4—9 mmol·L−1 during the stationary phase of growth with glucose as energy source. This value was dramatically reduced when pentoses were used as carbon source. Lactose and cellobiose, starch and cellulose were suitable substrates for the H2 production in some but not all isolates. No H2 was produced by proteinaceous substrate, such as blood. Results show that both substrate utilization and physiological responses (growth, excretion of carboxylates, H2 production) are unique functions of each isolate.

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Purification and characterization of an extracellular alpha-amylase from Clostridium perfringens type A

Cited by 25 publications

References 29 publications

Production, purification and characterization of α-amylase from Trichoderma harzianum grown on mandarin peel

Production, purification and characterization of α-amylase from Trichoderma harzianum grown on mandarin peel

Sporulation-promoting ability of Clostridium perfringens culture fluids

Carbon source utilization and hydrogen production by isolated anaerobic bacteria

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