Purification and characterisation of amylolytic enzymes from thermophilic fungus Thermomyces lanuginosus strain ATCC 34626

Q. Nguyen, Judit M. Rezessy-Szabó, Marc Claeyssens, Ingeborg Stals, Ágoston Hoschke

Research output: Contribution to journalArticle

114 Citations (Scopus)

Abstract

Amylolytic enzymes (α-amylase and glucoamylase) from Thermomyces lanuginosus ATCC 34626 were purified to electrophoretic homogeneity. The molecular mass of purified α-amylase and glucoamylase were 61 and 75kDa, respectively. Their pI values were calculated to be 3.5-3.6 and 4.1-4.3. The amylolytic enzymes from T. lanuginosus exhibit pH optima in the range 4.6-6.6 in the case of α-amylase and 4.4-5.6 in the case of glucoamylase. Both purified enzymes have temperature optima at 70°C. Zn2+ ions strongly inhibit both enzyme activities. Mn2+ and Fe2+ ions are activators in the case of glucoamylase; Ca2+ and Ba2+ are activators in the case of α-amylase. With half-life times longer than 1 day at 60°C both enzymes prove to be thermostable in the pH range 4.5-8.5. The amylolytic enzymes from T. lanuginosus loose activities rapidly when incubated at temperature higher than 80°C or at pH lower than 4.0. Both enzymes are found to be glycosylated; 8.5% carbohydrate in the case of α-amylase and 3.3% in the case of glucoamylase. The Km and Vmax of α-amylase on soluble starch were 0.68mg/ml and 45.19U/mg, respectively. The Km values of glucoamylase on maltose, maltotriose, maltotetraose, maltopentose and soluble starch were 6.5, 3.5, 2.1, 1.1mM and 0.8mg/ml, respectively. The first 37 residues of N-terminal of the purified α-amylase of T. lanuginosus ATCC 34626 were sequenced. Almost complete homology with the α-amylase from Aspergillus oryzae and Emericella nidulans was observed.

Original languageEnglish
Pages (from-to)345-352
Number of pages8
JournalEnzyme and Microbial Technology
Volume31
Issue number3
DOIs
Publication statusPublished - Aug 2 2002

Fingerprint

Amylases
Fungi
Glucan 1,4-alpha-Glucosidase
Purification
Enzymes
Starch
Emericella
Ions
Aspergillus oryzae
Maltose
Aspergillus nidulans
Temperature
Aspergillus
Enzyme activity
Molecular mass
Carbohydrates
Half-Life

Keywords

  • α-Amylase
  • Enzyme purification and kinetics
  • Glucoamylase
  • T. lanuginosus

ASJC Scopus subject areas

  • Biochemistry
  • Biotechnology
  • Applied Microbiology and Biotechnology

Cite this

Purification and characterisation of amylolytic enzymes from thermophilic fungus Thermomyces lanuginosus strain ATCC 34626. / Nguyen, Q.; Rezessy-Szabó, Judit M.; Claeyssens, Marc; Stals, Ingeborg; Hoschke, Ágoston.

In: Enzyme and Microbial Technology, Vol. 31, No. 3, 02.08.2002, p. 345-352.

Research output: Contribution to journalArticle

Nguyen, Q. ; Rezessy-Szabó, Judit M. ; Claeyssens, Marc ; Stals, Ingeborg ; Hoschke, Ágoston. / Purification and characterisation of amylolytic enzymes from thermophilic fungus Thermomyces lanuginosus strain ATCC 34626. In: Enzyme and Microbial Technology. 2002 ; Vol. 31, No. 3. pp. 345-352.
@article{efa30a87ab2741f99510c89f9c931fbc,
title = "Purification and characterisation of amylolytic enzymes from thermophilic fungus Thermomyces lanuginosus strain ATCC 34626",
abstract = "Amylolytic enzymes (α-amylase and glucoamylase) from Thermomyces lanuginosus ATCC 34626 were purified to electrophoretic homogeneity. The molecular mass of purified α-amylase and glucoamylase were 61 and 75kDa, respectively. Their pI values were calculated to be 3.5-3.6 and 4.1-4.3. The amylolytic enzymes from T. lanuginosus exhibit pH optima in the range 4.6-6.6 in the case of α-amylase and 4.4-5.6 in the case of glucoamylase. Both purified enzymes have temperature optima at 70°C. Zn2+ ions strongly inhibit both enzyme activities. Mn2+ and Fe2+ ions are activators in the case of glucoamylase; Ca2+ and Ba2+ are activators in the case of α-amylase. With half-life times longer than 1 day at 60°C both enzymes prove to be thermostable in the pH range 4.5-8.5. The amylolytic enzymes from T. lanuginosus loose activities rapidly when incubated at temperature higher than 80°C or at pH lower than 4.0. Both enzymes are found to be glycosylated; 8.5{\%} carbohydrate in the case of α-amylase and 3.3{\%} in the case of glucoamylase. The Km and Vmax of α-amylase on soluble starch were 0.68mg/ml and 45.19U/mg, respectively. The Km values of glucoamylase on maltose, maltotriose, maltotetraose, maltopentose and soluble starch were 6.5, 3.5, 2.1, 1.1mM and 0.8mg/ml, respectively. The first 37 residues of N-terminal of the purified α-amylase of T. lanuginosus ATCC 34626 were sequenced. Almost complete homology with the α-amylase from Aspergillus oryzae and Emericella nidulans was observed.",
keywords = "α-Amylase, Enzyme purification and kinetics, Glucoamylase, T. lanuginosus",
author = "Q. Nguyen and Rezessy-Szab{\'o}, {Judit M.} and Marc Claeyssens and Ingeborg Stals and {\'A}goston Hoschke",
year = "2002",
month = "8",
day = "2",
doi = "10.1016/S0141-0229(02)00128-X",
language = "English",
volume = "31",
pages = "345--352",
journal = "Enzyme and Microbial Technology",
issn = "0141-0229",
publisher = "Elsevier Inc.",
number = "3",

}

TY - JOUR

T1 - Purification and characterisation of amylolytic enzymes from thermophilic fungus Thermomyces lanuginosus strain ATCC 34626

AU - Nguyen, Q.

AU - Rezessy-Szabó, Judit M.

AU - Claeyssens, Marc

AU - Stals, Ingeborg

AU - Hoschke, Ágoston

PY - 2002/8/2

Y1 - 2002/8/2

N2 - Amylolytic enzymes (α-amylase and glucoamylase) from Thermomyces lanuginosus ATCC 34626 were purified to electrophoretic homogeneity. The molecular mass of purified α-amylase and glucoamylase were 61 and 75kDa, respectively. Their pI values were calculated to be 3.5-3.6 and 4.1-4.3. The amylolytic enzymes from T. lanuginosus exhibit pH optima in the range 4.6-6.6 in the case of α-amylase and 4.4-5.6 in the case of glucoamylase. Both purified enzymes have temperature optima at 70°C. Zn2+ ions strongly inhibit both enzyme activities. Mn2+ and Fe2+ ions are activators in the case of glucoamylase; Ca2+ and Ba2+ are activators in the case of α-amylase. With half-life times longer than 1 day at 60°C both enzymes prove to be thermostable in the pH range 4.5-8.5. The amylolytic enzymes from T. lanuginosus loose activities rapidly when incubated at temperature higher than 80°C or at pH lower than 4.0. Both enzymes are found to be glycosylated; 8.5% carbohydrate in the case of α-amylase and 3.3% in the case of glucoamylase. The Km and Vmax of α-amylase on soluble starch were 0.68mg/ml and 45.19U/mg, respectively. The Km values of glucoamylase on maltose, maltotriose, maltotetraose, maltopentose and soluble starch were 6.5, 3.5, 2.1, 1.1mM and 0.8mg/ml, respectively. The first 37 residues of N-terminal of the purified α-amylase of T. lanuginosus ATCC 34626 were sequenced. Almost complete homology with the α-amylase from Aspergillus oryzae and Emericella nidulans was observed.

AB - Amylolytic enzymes (α-amylase and glucoamylase) from Thermomyces lanuginosus ATCC 34626 were purified to electrophoretic homogeneity. The molecular mass of purified α-amylase and glucoamylase were 61 and 75kDa, respectively. Their pI values were calculated to be 3.5-3.6 and 4.1-4.3. The amylolytic enzymes from T. lanuginosus exhibit pH optima in the range 4.6-6.6 in the case of α-amylase and 4.4-5.6 in the case of glucoamylase. Both purified enzymes have temperature optima at 70°C. Zn2+ ions strongly inhibit both enzyme activities. Mn2+ and Fe2+ ions are activators in the case of glucoamylase; Ca2+ and Ba2+ are activators in the case of α-amylase. With half-life times longer than 1 day at 60°C both enzymes prove to be thermostable in the pH range 4.5-8.5. The amylolytic enzymes from T. lanuginosus loose activities rapidly when incubated at temperature higher than 80°C or at pH lower than 4.0. Both enzymes are found to be glycosylated; 8.5% carbohydrate in the case of α-amylase and 3.3% in the case of glucoamylase. The Km and Vmax of α-amylase on soluble starch were 0.68mg/ml and 45.19U/mg, respectively. The Km values of glucoamylase on maltose, maltotriose, maltotetraose, maltopentose and soluble starch were 6.5, 3.5, 2.1, 1.1mM and 0.8mg/ml, respectively. The first 37 residues of N-terminal of the purified α-amylase of T. lanuginosus ATCC 34626 were sequenced. Almost complete homology with the α-amylase from Aspergillus oryzae and Emericella nidulans was observed.

KW - α-Amylase

KW - Enzyme purification and kinetics

KW - Glucoamylase

KW - T. lanuginosus

UR - http://www.scopus.com/inward/record.url?scp=0037008421&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0037008421&partnerID=8YFLogxK

U2 - 10.1016/S0141-0229(02)00128-X

DO - 10.1016/S0141-0229(02)00128-X

M3 - Article

AN - SCOPUS:0037008421

VL - 31

SP - 345

EP - 352

JO - Enzyme and Microbial Technology

JF - Enzyme and Microbial Technology

SN - 0141-0229

IS - 3

ER -