Endoxylanase II from Trichoderma reesei has several isoforms with different isoelectric points

Arja Lappalainen (Corresponding Author), Matti Siika-aho, Nisse Kalkkinen, Richard Fagerström, Maija Tenkanen

Research output: Contribution to journalArticleScientificpeer-review

26 Citations (Scopus)

Abstract

Two minor xylanases present in Trichoderma reesei Rut C30 cultivation broth were purified as a mixture using ion‐exchange, hydrophobic‐interaction and gel chromatography. The purified enzyme preparation contained two active xylanases with pI values of 7.1 and 8.1. Both components had a molecular mass of 20 kDa. The purified xylanase preparation exhibited properties very similar to those of the previously isolated XYL II (pI 9.0) of T. reesei Rut C30. The activity and stability properties, apparent kinetic parameters as well as the titration curve forms were similar. The major difference in enzymic properties was the significantly lower specific activity of the pI‐7.1+8.1 xylanase mixture (3350 nkat/mg) compared with the specific activity of XYL II (13500 nkat/mg). Amino acid sequences of tryptic peptides (34% of the total amino acid sequence was determined) were identical to the amino acid sequence of XYL II. Furthermore, in vitro modification of the pI‐9.0 form of XYL II to pI‐8.1 and pI‐7.1 forms was demonstrated. Thus the purified xylanase preparation most probably contained two modified forms of XYL II. The primary amino acid sequence of XYL II contains 28 glutamine and asparagine residues and theoretically deamination of one of them lowers the pI to 8.06 and deamination of two amino acids lowers the pI to 7.02.

Original languageEnglish
Pages (from-to)61 - 68
Number of pages8
JournalBiotechnology and Applied Biochemistry
Volume31
Issue number1
DOIs
Publication statusPublished - 2000
MoE publication typeA1 Journal article-refereed

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Endo-1,4-beta Xylanases
Trichoderma
Isoelectric Point
Amino acids
Amino Acid Sequence
Protein Isoforms
Amino Acids
Deamination
Asparagine
Glutamine
Gel Chromatography
Molecular mass
Chromatography
Titration
Kinetic parameters
Peptides
Gels
Enzymes

Cite this

@article{d69011d59e214c638c493d5f5e5fa25c,
title = "Endoxylanase II from Trichoderma reesei has several isoforms with different isoelectric points",
abstract = "Two minor xylanases present in Trichoderma reesei Rut C30 cultivation broth were purified as a mixture using ion‐exchange, hydrophobic‐interaction and gel chromatography. The purified enzyme preparation contained two active xylanases with pI values of 7.1 and 8.1. Both components had a molecular mass of 20 kDa. The purified xylanase preparation exhibited properties very similar to those of the previously isolated XYL II (pI 9.0) of T. reesei Rut C30. The activity and stability properties, apparent kinetic parameters as well as the titration curve forms were similar. The major difference in enzymic properties was the significantly lower specific activity of the pI‐7.1+8.1 xylanase mixture (3350 nkat/mg) compared with the specific activity of XYL II (13500 nkat/mg). Amino acid sequences of tryptic peptides (34{\%} of the total amino acid sequence was determined) were identical to the amino acid sequence of XYL II. Furthermore, in vitro modification of the pI‐9.0 form of XYL II to pI‐8.1 and pI‐7.1 forms was demonstrated. Thus the purified xylanase preparation most probably contained two modified forms of XYL II. The primary amino acid sequence of XYL II contains 28 glutamine and asparagine residues and theoretically deamination of one of them lowers the pI to 8.06 and deamination of two amino acids lowers the pI to 7.02.",
author = "Arja Lappalainen and Matti Siika-aho and Nisse Kalkkinen and Richard Fagerstr{\"o}m and Maija Tenkanen",
year = "2000",
doi = "10.1042/BA19990066",
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Endoxylanase II from Trichoderma reesei has several isoforms with different isoelectric points. / Lappalainen, Arja (Corresponding Author); Siika-aho, Matti; Kalkkinen, Nisse; Fagerström, Richard; Tenkanen, Maija.

In: Biotechnology and Applied Biochemistry, Vol. 31, No. 1, 2000, p. 61 - 68.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Endoxylanase II from Trichoderma reesei has several isoforms with different isoelectric points

AU - Lappalainen, Arja

AU - Siika-aho, Matti

AU - Kalkkinen, Nisse

AU - Fagerström, Richard

AU - Tenkanen, Maija

PY - 2000

Y1 - 2000

N2 - Two minor xylanases present in Trichoderma reesei Rut C30 cultivation broth were purified as a mixture using ion‐exchange, hydrophobic‐interaction and gel chromatography. The purified enzyme preparation contained two active xylanases with pI values of 7.1 and 8.1. Both components had a molecular mass of 20 kDa. The purified xylanase preparation exhibited properties very similar to those of the previously isolated XYL II (pI 9.0) of T. reesei Rut C30. The activity and stability properties, apparent kinetic parameters as well as the titration curve forms were similar. The major difference in enzymic properties was the significantly lower specific activity of the pI‐7.1+8.1 xylanase mixture (3350 nkat/mg) compared with the specific activity of XYL II (13500 nkat/mg). Amino acid sequences of tryptic peptides (34% of the total amino acid sequence was determined) were identical to the amino acid sequence of XYL II. Furthermore, in vitro modification of the pI‐9.0 form of XYL II to pI‐8.1 and pI‐7.1 forms was demonstrated. Thus the purified xylanase preparation most probably contained two modified forms of XYL II. The primary amino acid sequence of XYL II contains 28 glutamine and asparagine residues and theoretically deamination of one of them lowers the pI to 8.06 and deamination of two amino acids lowers the pI to 7.02.

AB - Two minor xylanases present in Trichoderma reesei Rut C30 cultivation broth were purified as a mixture using ion‐exchange, hydrophobic‐interaction and gel chromatography. The purified enzyme preparation contained two active xylanases with pI values of 7.1 and 8.1. Both components had a molecular mass of 20 kDa. The purified xylanase preparation exhibited properties very similar to those of the previously isolated XYL II (pI 9.0) of T. reesei Rut C30. The activity and stability properties, apparent kinetic parameters as well as the titration curve forms were similar. The major difference in enzymic properties was the significantly lower specific activity of the pI‐7.1+8.1 xylanase mixture (3350 nkat/mg) compared with the specific activity of XYL II (13500 nkat/mg). Amino acid sequences of tryptic peptides (34% of the total amino acid sequence was determined) were identical to the amino acid sequence of XYL II. Furthermore, in vitro modification of the pI‐9.0 form of XYL II to pI‐8.1 and pI‐7.1 forms was demonstrated. Thus the purified xylanase preparation most probably contained two modified forms of XYL II. The primary amino acid sequence of XYL II contains 28 glutamine and asparagine residues and theoretically deamination of one of them lowers the pI to 8.06 and deamination of two amino acids lowers the pI to 7.02.

U2 - 10.1042/BA19990066

DO - 10.1042/BA19990066

M3 - Article

VL - 31

SP - 61

EP - 68

JO - Biotechnology and Applied Biochemistry

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SN - 0885-4513

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