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Literature summary extracted from

  • Ben Abdelmalek-Khedher, I.; Urdaci, M.C.; Limam, F.; Schmitter, J.M.; Marzouki, M.N.; Bressollier, P.
    Purification, characterization, and partial primary sequence of a major-maltotriose-producing alpha-amylase, ScAmy43, from Sclerotinia sclerotiorum (2008), J. Microbiol. Biotechnol., 18, 1555-1563.
    View publication on PubMed

Activating Compound

EC Number Activating Compound Comment Organism Structure
3.2.1.1 2-mercaptoethanol 5 mM, pH 4.0, 55°C, 172% relative activity, strongly increase the activity, probably due to a better accessibility of the substrate to the catalytic site after disruption of the intersulfide bridge Sclerotinia sclerotiorum
3.2.1.1 dithiothreitol 5 mM, pH 4.0, 55°C, 166% relative activity, strongly increase the activity, probably due to a better accessibility of the substrate to the catalytic site after disruption of the intersulfide bridge Sclerotinia sclerotiorum

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.2.1.1 BaCl2 5 mM, pH 4.0, 55°C, 25% relative activity Sclerotinia sclerotiorum
3.2.1.1 CuCl2 5 mM, pH 4.0, 55°C, 26% relative activity Sclerotinia sclerotiorum
3.2.1.1 EDTA 5, 10, and 20 mM, pH 4.0, 55°C, 95% relative activity, enzyme is not sensitive to chelating agent, probably not a metalloenzyme Sclerotinia sclerotiorum
3.2.1.1 EGTA 5, 10, and 20 mM, pH 4.0, 55°C, 100% relative activity, enzyme is not sensitive to chelating agent, probably not a metalloenzyme Sclerotinia sclerotiorum
3.2.1.1 FeCl2 5 mM, pH 4.0, 55°C, 85% relative activity Sclerotinia sclerotiorum
3.2.1.1 MnCl2 5 mM, pH 4.0, 55°C, 44% relative activity Sclerotinia sclerotiorum
3.2.1.1 SDS 2%, pH 4.0, 55°C, 11% relative activity Sclerotinia sclerotiorum
3.2.1.1 Urea 5 M, pH 4.0, 55°C, 63% relative activity Sclerotinia sclerotiorum

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.2.1.1 additional information
-
 soluble starch Km: 1.66 mg/ml, Vmax: 0.0001 mmol/min/ml Sclerotinia sclerotiorum

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.2.1.1 CaCl2 5 mM, pH 4.0, 55°C, 111% relative activity Sclerotinia sclerotiorum
3.2.1.1 MgCl2 5 mM, pH 4.0, 55°C, 103% relative activity Sclerotinia sclerotiorum
3.2.1.1 ZnCl2 5 mM, pH 4.0, 55°C, 104% relative activity Sclerotinia sclerotiorum

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.2.1.1 40000
-
 gel filtration Sclerotinia sclerotiorum
3.2.1.1 43000
-
 SDS-PAGE Sclerotinia sclerotiorum

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.1 Sclerotinia sclerotiorum
-
 pathogenic fungus, grown on oats flour
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.1.1
-
 Sclerotinia sclerotiorum

Source Tissue

EC Number Source Tissue Comment Organism Textmining
3.2.1.1 culture medium
-
 Sclerotinia sclerotiorum
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.1 additional information ScAmy43 does not hydrolyze native starches Sclerotinia sclerotiorum ?
-
 ?
3.2.1.1 starch + H2O pH 4, 55°C, various starches are tested: soluble potato starch as reference 100% relative activity, gelatinized amylose: 74%, potato native starch: 0.2%, potato gelatinized starch: 106%, maize native starch: 0.8%, maize gelatinized starch: 53%, rice native starch: 0.5%, and rice gelatinized starch: 52% Sclerotinia sclerotiorum maltotriose + glucose + maltose + maltotetraose identified by thin-layer-chromatography, maltotriose represents 70% of the end products, only traces of glucose, small amounts of maltose and maltotetraose ?

Subunits

EC Number Subunits Comment Organism
3.2.1.1 monomer 1 * 43000, SDS-PAGE Sclerotinia sclerotiorum

Synonyms

EC Number Synonyms Comment Organism
3.2.1.1 maltotriose-producing alpha-amylase
-
 Sclerotinia sclerotiorum
3.2.1.1 ScAmy43
-
 Sclerotinia sclerotiorum

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.2.1.1 55
-
 assay at Sclerotinia sclerotiorum

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
3.2.1.1 40 65 40°C: ca. 40% relative activity, 65°C: ca. 40% relative activity, pH 4.0 for 30 min Sclerotinia sclerotiorum

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.2.1.1 40
-
 stable in the absence of substrate during 30 min pre-incubation, assay at pH 4.0 and 55°C, above 40°C rapid inactivation Sclerotinia sclerotiorum
3.2.1.1 50
-
 in the presence of 1% (w/v) potato starch the thermostability is enhanced above 50°C Sclerotinia sclerotiorum
3.2.1.1 55
-
 20% of maximal activity after 30 min pre-incubation in the absence of a substrate, but the enzyme is stable during short time assay for 30 min Sclerotinia sclerotiorum

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.2.1.1 4
-
-
 Sclerotinia sclerotiorum

pH Range

EC Number pH Minimum pH Maximum Comment Organism
3.2.1.1 3 6.5 pH 3: below 10%, pH 6.5: ca. 10 relativ activity, 50% of maximal activity at pH 3.5 and 5.5, 55°C Sclerotinia sclerotiorum

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
3.2.1.1 4 7 ScAmy43 is stable over a narrow range, pH 4.0: below 60% residual activity, pH 7: below 80% residual activity, after incubation at various pH at 4°C for 24 h, assay at pH 4.0 and 55°C for 30 min Sclerotinia sclerotiorum