Purification of α-Amylase Isoenzymes from Scytalidium thermophilum on a Fluidized Bed of Alginate Beads Followed by Concanavalin A–Agarose Column Chromatography

Roy, Ipsita; Sastry, Murali; Johri, B. N.; Gupta, Munishwar N.

doi:10.1006/prep.2000.1308

Cited by 14 publications

(2 citation statements)

References 31 publications

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“…A Sephacryl S-200 column was used to obtain A3 with the highest possible specific activities of 2260 units/mg protein which represented 4.47 fold purification over the cell-free broth with 39.4% recovery (Figure 4). Similar specific activity of -amylases from Scytalidium thermophilum (2450 units/ mg protein) was detected (Roy et al, 2000). The lowest specific activity of -amylases from Acremonium sporosulcatum (55.35 units/ mg protein) (Valaparla, 2010) and Thermobifida fusca (245 units/ mg mg protein) (Yang and Liu, 2004) Molecular weights of α-amylases vary from about 10 to 210 kDa.…”

Section: Purification Of α-Amylase From T Harzianummentioning

confidence: 59%

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: Purification Of α-Amylase From T Harzianummentioning

confidence: 59%

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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“…Many publications reported the affinity of amylases (from various sources) for alginate [28,29]. This character was used in the separation and purification of amylases [30]. Very few publications have been found addressing the covalent immobilization of enzymes onto alginate [22,23,31,32].…”

mentioning

confidence: 99%

Affinity Covalent Immobilization of Glucoamylase onto ρ-Benzoquinone Activated Alginate Beads: I. Beads Preparation and Characterization

Eldin

Seuror

Nasr

et al. 2010

Appl Biochem Biotechnol

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ρ-Benzoquinone-activated alginate beads were presented as a new carrier for affinity covalent immobilization of glucoamylase enzyme. Evidences of alginate modification were extracted from FT-IR and thermal gravimetric analysis and supported by morphological changes recognized through SEM examination. Factors affecting the modification process such as ρ-benzoquinone (PBQ) concentration, reaction time, reaction temperature, reaction pH and finally alginate concentration, have been studied. Its influence on the amount of coupled PBQ was consequently correlated to the changes of the catalytic activity and the retained activity of immobilized enzyme, the main parameters judging the success of the immobilization process. The immobilized glucoamylase was found kept almost 80% of its native activity giving proof of non-significant substrate, starch, diffusion limitation. The proposed affinity covalent immobilizing technique would rank among the potential strategies for efficient immobilization of glucoamylase enzyme.

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Free Polymeric Bioligands in Aqueous Two-Phase Affinity Extractions of Microbial Xylanases and Pullulanase

Teotia

Lata

Gupta

2001

Protein Expression and Purification

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Purification of α-Amylase Isoenzymes from Scytalidium thermophilum on a Fluidized Bed of Alginate Beads Followed by Concanavalin A–Agarose Column Chromatography

Cited by 14 publications

References 31 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

Affinity Covalent Immobilization of Glucoamylase onto ρ-Benzoquinone Activated Alginate Beads: I. Beads Preparation and Characterization

Free Polymeric Bioligands in Aqueous Two-Phase Affinity Extractions of Microbial Xylanases and Pullulanase

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