Purification and properties of a maltoheptaose- and maltohexaose-forming amylase produced by Bacillus subtilis US116

Messaoud, Ezzedine Ben; Ali, Mamdouh Ben; Elleuch, N.; Masmoudi, Najla; Béjar, Samír

doi:10.1016/j.enzmictec.2004.03.002

Cited by 36 publications

(25 citation statements)

References 19 publications

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“…Endo and exo-hydrolytic activities were assayed, as previously reported [24] according to Ceralpha and Betamyl methods, respectively, using the corresponding assay kits commercialized by Megazyme (Wicklow, Ireland).…”

Section: Endo and Exo-hydrolytic Activities Assaymentioning

confidence: 99%

Cloning and Sequencing of an Original Gene Encoding a Maltogenic Amylase from Bacillus sp. US149 Strain and Characterization of the Recombinant Activity

et al. 2007

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The molecular diagnosis of prion diseases almost always involves the use of a protease to distinguish PrPC from PrPSc and invariably the protease of choice is proteinase K. Here, we have applied the protease thermolysin to the diagnosis of animal prion diseases. This thermostable protease cleaves at the hydrophobic residues Leu, Ile, Phe, Val, Ala, and Met, residues that are absent from the protease accessible aminoterminal region of PrPSc. Therefore, although thermolysin readily digests PrPC into small protein fragments, full-length PrPSc is resistant to such proteolysis. This contrasts with proteinase K digestion where an aminoterminally truncated PrPSc species is produced, PrP27-30. Thermolysin was used in the diagnosis of ovine scrapie and bovine spongiform encephalopathy and produced comparable assay sensitivity to assays using proteinase K digestion. Furthermore, we demonstrated the concentration of thermolysin-resistant PrPSc using immobilized metal-affinity chromatography. The use of thermolysin to reveal a full-length PrPSc has application for the development of novel immunodiagnostics by exploiting the wide range of commercially available immunoreagents and metal affinity matrices that bind the amino-terminal region of PrP. In addition, thermolysin provides a complementary tool to proteinase K to allow the study of the contribution of the amino-terminal domain of PrPSc to disease pathogenesis.

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Section: Endo and Exo-hydrolytic Activities Assaymentioning

confidence: 99%

Cloning and Sequencing of an Original Gene Encoding a Maltogenic Amylase from Bacillus sp. US149 Strain and Characterization of the Recombinant Activity

et al. 2007

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“…Furthermore, it is worthy to note that these enzymes performed some important difference in enzymatic properties. Firstly, besides being active and stable at low pH, the enzyme BYXA indicated remarkable stability in a wide range of pH values BYXA, however, the B. amyloliquefaciens -amylase produced largely maltose, maltotriose, maltopentaose and maltohexaose from soluble starch (Demirkan et al, 2005) while the B. subtilis -amylase yielded predominantly maltohexaose and maltoheptaose (Ben Messaoud et al, 2004). These strongly demonstrated a relationship between the primary sequence and catalysis function of the protein.…”

Section: Resultsmentioning

confidence: 98%

Molecular cloning and characterization of an α-amylase with raw starch digestibility fromBacillus sp. YX-1

Liu

2009

Ann. Microbiol.

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The complete gene (amy) that encoding a raw starch digesting -amylase (BYXA) from Bacillus sp. YX-1 was cloned, sequenced and expressed in Escherichia coli. The amy gene consisted of 1545 base pairs and encoded a protein of 514 amino acid residues with a calculated molecular mass of 58 kDa. The recombinant protein was purified by Ni 2+ affinity chromatography and subsequently characterized. The optimal pH and temperature for the purified recombinant enzyme were 5.5 and 45 °C, respectively. The enzyme showed stability over a pH range of 5.0-11.0 and below 50 °C. The enzyme exhibited remarkable digestibility towards raw starch at low pH values, which was valuable in starch processing industry.

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“…However, amylases that speciWcally produce malto-oligosaccharides from starch have been reported. These include the maltohexaose producing amylases of Aerobacter and Bacillus [2][3][4], a maltopentaose-producing amylase from Bacillus [5], the maltotetraose-producing amylases from Pseudomonas [6], Bacillus [7] and ChloroXexus [8] and the maltotriose producing amylases from Streptomyces [9], Bacillus [10], Microbacterium [11], Natronoccus [12] and Streptococcus [13,14].…”

Section: Introductionmentioning

confidence: 99%

Cloning and characterization of a maltotriose-producing α-amylase gene from Thermobifida fusca

Yang¹,

Liu²

2007

J Ind Microbiol Biotechnol

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The gene (tfa), encoding a maltotriose-producing alpha-amylase from Thermobifida fusca NTU22, was cloned, sequenced and expressed in Escherichia coli. The gene consists of 1,815 base pairs and encodes a protein of 605 amino acids. The base composition of the tfa coding sequence is 69% G+C and the protein has a predicted pI value of 5.5. The deduced amino acid sequence of the tfa amylase exhibited a high degree of similarity with amylases from Thermomonospora curvata and Streptomyces amylases. The purified amylase could be detected as a single band of about 65 kDa by SDS-polyacrylamide gel electrophoresis and this agrees with the predicted size based on the nucleotide sequence. The optimal pH and temperature of the purified amylase were 7.0 and 60 degrees C, respectively. The properties of purified amylase from the E. coli transformant are similar to that of an amylase purified from the original T. fusca NTU22.

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Purification and properties of a maltoheptaose- and maltohexaose-forming amylase produced by Bacillus subtilis US116

Cited by 36 publications

References 19 publications

Cloning and Sequencing of an Original Gene Encoding a Maltogenic Amylase from Bacillus sp. US149 Strain and Characterization of the Recombinant Activity

Cloning and Sequencing of an Original Gene Encoding a Maltogenic Amylase from Bacillus sp. US149 Strain and Characterization of the Recombinant Activity

Molecular cloning and characterization of an α-amylase with raw starch digestibility fromBacillus sp. YX-1

Cloning and characterization of a maltotriose-producing α-amylase gene from Thermobifida fusca

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