Alfa-Galactosidases of Penicillium simplicissimum: production, purification and characterization of the gene encoding AGLI

Elina Luonteri, Edward Alatalo, Matti Siika-aho, Merja Penttilä, Maija Tenkanen

Research output: Contribution to journalArticleScientificpeer-review

Abstract

Production of extracellular a‐galactosidases by the filamentous fungus Penicillium simplicissimum (previously P. janthinellum) VTT‐D‐78090 was studied on different carbon sources.
Steam‐exploded oat husks were chosen as the best carbon source for enzyme production. Three a‐galactosidases (AGL) were purified from the culture filtrate using ion‐exchange chromatography, hydrophobic interaction chromatography and gel filtration.
The isoelectric points of AGLI, AGLII and AGLIII were 5.2, 4.4 and 7.0, and the molecular masses as determined by SDS/PAGE were 61, 84 and 61 kDa, respectively. All enzymes were glycosylated. The optimum pH for the activity of AGLI and AGLIII was between 3.0 and 4.5 and that of AGLII was between 4.0 and 5.0. AGLII was more stable and more resistant to product inhibition by galactose than the other two enzymes. AGLI and AGLIII were also inhibited by p‐nitrophenol‐a‐D‐galactopyranoside, the substrate used for enzyme activity assay.
The gene encoding AGLI was cloned and sequenced. The gene, agl1, encodes 435 amino acids including the signal sequence. It showed similarity with the other a‐galactosidases belonging to the glycosyl hydrolase family 27.
The N‐terminal amino acid sequence of AGLIII was also similar to the sequences of other members of family 27, whereas the N‐terminus of AGLII was completely different from the sequences of other reported hydrolases.
Original languageEnglish
Pages (from-to)179-188
JournalBiotechnology and Applied Biochemistry
Volume28
Issue number2
DOIs
Publication statusPublished - 1998
MoE publication typeA1 Journal article-refereed

Fingerprint

alpha-Galactosidase
Gene encoding
Penicillium
Purification
Hydrolases
Enzymes
Hydrophobic chromatography
Amino acids
Carbon
Genes
Enzyme inhibition
Amino Acids
Isoelectric Point
Enzyme Assays
Enzyme activity
Molecular mass
Protein Sorting Signals
Chromatography
Fungi
Galactose

Cite this

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title = "Alfa-Galactosidases of Penicillium simplicissimum: production, purification and characterization of the gene encoding AGLI",
abstract = "Production of extracellular a‐galactosidases by the filamentous fungus Penicillium simplicissimum (previously P. janthinellum) VTT‐D‐78090 was studied on different carbon sources.Steam‐exploded oat husks were chosen as the best carbon source for enzyme production. Three a‐galactosidases (AGL) were purified from the culture filtrate using ion‐exchange chromatography, hydrophobic interaction chromatography and gel filtration. The isoelectric points of AGLI, AGLII and AGLIII were 5.2, 4.4 and 7.0, and the molecular masses as determined by SDS/PAGE were 61, 84 and 61 kDa, respectively. All enzymes were glycosylated. The optimum pH for the activity of AGLI and AGLIII was between 3.0 and 4.5 and that of AGLII was between 4.0 and 5.0. AGLII was more stable and more resistant to product inhibition by galactose than the other two enzymes. AGLI and AGLIII were also inhibited by p‐nitrophenol‐a‐D‐galactopyranoside, the substrate used for enzyme activity assay. The gene encoding AGLI was cloned and sequenced. The gene, agl1, encodes 435 amino acids including the signal sequence. It showed similarity with the other a‐galactosidases belonging to the glycosyl hydrolase family 27. The N‐terminal amino acid sequence of AGLIII was also similar to the sequences of other members of family 27, whereas the N‐terminus of AGLII was completely different from the sequences of other reported hydrolases.",
author = "Elina Luonteri and Edward Alatalo and Matti Siika-aho and Merja Penttil{\"a} and Maija Tenkanen",
year = "1998",
doi = "10.1111/j.1470-8744.1998.tb00528.x",
language = "English",
volume = "28",
pages = "179--188",
journal = "Biotechnology and Applied Biochemistry",
issn = "0885-4513",
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}

Alfa-Galactosidases of Penicillium simplicissimum: production, purification and characterization of the gene encoding AGLI. / Luonteri, Elina; Alatalo, Edward; Siika-aho, Matti; Penttilä, Merja; Tenkanen, Maija.

In: Biotechnology and Applied Biochemistry, Vol. 28, No. 2, 1998, p. 179-188.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Alfa-Galactosidases of Penicillium simplicissimum: production, purification and characterization of the gene encoding AGLI

AU - Luonteri, Elina

AU - Alatalo, Edward

AU - Siika-aho, Matti

AU - Penttilä, Merja

AU - Tenkanen, Maija

PY - 1998

Y1 - 1998

N2 - Production of extracellular a‐galactosidases by the filamentous fungus Penicillium simplicissimum (previously P. janthinellum) VTT‐D‐78090 was studied on different carbon sources.Steam‐exploded oat husks were chosen as the best carbon source for enzyme production. Three a‐galactosidases (AGL) were purified from the culture filtrate using ion‐exchange chromatography, hydrophobic interaction chromatography and gel filtration. The isoelectric points of AGLI, AGLII and AGLIII were 5.2, 4.4 and 7.0, and the molecular masses as determined by SDS/PAGE were 61, 84 and 61 kDa, respectively. All enzymes were glycosylated. The optimum pH for the activity of AGLI and AGLIII was between 3.0 and 4.5 and that of AGLII was between 4.0 and 5.0. AGLII was more stable and more resistant to product inhibition by galactose than the other two enzymes. AGLI and AGLIII were also inhibited by p‐nitrophenol‐a‐D‐galactopyranoside, the substrate used for enzyme activity assay. The gene encoding AGLI was cloned and sequenced. The gene, agl1, encodes 435 amino acids including the signal sequence. It showed similarity with the other a‐galactosidases belonging to the glycosyl hydrolase family 27. The N‐terminal amino acid sequence of AGLIII was also similar to the sequences of other members of family 27, whereas the N‐terminus of AGLII was completely different from the sequences of other reported hydrolases.

AB - Production of extracellular a‐galactosidases by the filamentous fungus Penicillium simplicissimum (previously P. janthinellum) VTT‐D‐78090 was studied on different carbon sources.Steam‐exploded oat husks were chosen as the best carbon source for enzyme production. Three a‐galactosidases (AGL) were purified from the culture filtrate using ion‐exchange chromatography, hydrophobic interaction chromatography and gel filtration. The isoelectric points of AGLI, AGLII and AGLIII were 5.2, 4.4 and 7.0, and the molecular masses as determined by SDS/PAGE were 61, 84 and 61 kDa, respectively. All enzymes were glycosylated. The optimum pH for the activity of AGLI and AGLIII was between 3.0 and 4.5 and that of AGLII was between 4.0 and 5.0. AGLII was more stable and more resistant to product inhibition by galactose than the other two enzymes. AGLI and AGLIII were also inhibited by p‐nitrophenol‐a‐D‐galactopyranoside, the substrate used for enzyme activity assay. The gene encoding AGLI was cloned and sequenced. The gene, agl1, encodes 435 amino acids including the signal sequence. It showed similarity with the other a‐galactosidases belonging to the glycosyl hydrolase family 27. The N‐terminal amino acid sequence of AGLIII was also similar to the sequences of other members of family 27, whereas the N‐terminus of AGLII was completely different from the sequences of other reported hydrolases.

U2 - 10.1111/j.1470-8744.1998.tb00528.x

DO - 10.1111/j.1470-8744.1998.tb00528.x

M3 - Article

VL - 28

SP - 179

EP - 188

JO - Biotechnology and Applied Biochemistry

JF - Biotechnology and Applied Biochemistry

SN - 0885-4513

IS - 2

ER -