Role of the hga1-encoded extracellular beta-galactosidase of Hypocrea jecorina in lactose metabolism and cellulase induction

Bernhard Seiboth, Lukas Hartl, Noora Salovuori, Jari Vehmaanperä, Merja Penttilä, Christian Kubicek

Research output: Chapter in Book/Report/Conference proceedingConference abstract in proceedingsScientific

Abstract

Lactose is the only economic carbon source for protein production under the control of cellulase promoters by the ascomycete H. jecorina (anamorph: Trichoderma reesei). However, the mechanism by which lactose triggers cellulase formation is not understood. We have investigated the role of beta-galactosidase in lactose metabolism and cellulase induction in H. jecorina. A genomic copy of the bga1 gene predicting a protein with a MW of 111 kDa (incl. a signal sequence) was cloned using degenerate primers. The Bga1 belongs to the Glycosyl hydrolases family 35. Transcriptional analysis of the bga1 expression shows that it is highly expressed on L-arabinose, D-galactose and lower on lactose. Deletion of the gene showed that it is not essential for growth on lactose but that knock-out strains grew slower and produced about half the biomass of the wt strain, whereas amplification of the bga1 gene under the pki1 promoter resulted in faster growth with a reduced lag phase. Bga1 is not necessary for lactose-dependent induction of cbh1 gene expression in H. jecorina but its overexpression impairs lactose-induction of cbh1 expression. Analysis of the deletion strain further showed that bga1 encodes the major extracellular beta-galactosidase which is also partially cell-wall bound. An additional cell wall-bound beta-galactosidase activity could be shown but no evidence for an intracellular beta-galactosidase was obtained.
Original languageEnglish
Title of host publication22nd Fungal Genetics Conference at Asilomar
Subtitle of host publicationAbstracts from the 2003 Fungal Genetics Conference at Asilomar
DOIs
Publication statusPublished - 2003
Event22nd Fungal Genetics Conference - Asilomar, United States
Duration: 18 Mar 200322 Mar 2003

Publication series

SeriesFungal Genetics Reports
NumberArticle 18
Volume50

Conference

Conference22nd Fungal Genetics Conference
CountryUnited States
CityAsilomar
Period18/03/0322/03/03

Fingerprint

Trichoderma reesei
beta-galactosidase
endo-1,4-beta-glucanase
lactose
metabolism
promoter regions
cell walls
gene deletion
glycosidases
anamorphs
arabinose
signal peptide
Ascomycota
galactose
protein sources
polymerase chain reaction
genomics
economics
gene expression
carbon

Cite this

Seiboth, B., Hartl, L., Salovuori, N., Vehmaanperä, J., Penttilä, M., & Kubicek, C. (2003). Role of the hga1-encoded extracellular beta-galactosidase of Hypocrea jecorina in lactose metabolism and cellulase induction. In 22nd Fungal Genetics Conference at Asilomar: Abstracts from the 2003 Fungal Genetics Conference at Asilomar [424] Fungal Genetics Reports, No. Article 18, Vol.. 50 https://doi.org/10.4148/1941-4765.1164
Seiboth, Bernhard ; Hartl, Lukas ; Salovuori, Noora ; Vehmaanperä, Jari ; Penttilä, Merja ; Kubicek, Christian. / Role of the hga1-encoded extracellular beta-galactosidase of Hypocrea jecorina in lactose metabolism and cellulase induction. 22nd Fungal Genetics Conference at Asilomar: Abstracts from the 2003 Fungal Genetics Conference at Asilomar. 2003. (Fungal Genetics Reports; No. Article 18, Vol. 50).
@inbook{6a1b966ecc74483ba06a78a7c12c34be,
title = "Role of the hga1-encoded extracellular beta-galactosidase of Hypocrea jecorina in lactose metabolism and cellulase induction",
abstract = "Lactose is the only economic carbon source for protein production under the control of cellulase promoters by the ascomycete H. jecorina (anamorph: Trichoderma reesei). However, the mechanism by which lactose triggers cellulase formation is not understood. We have investigated the role of beta-galactosidase in lactose metabolism and cellulase induction in H. jecorina. A genomic copy of the bga1 gene predicting a protein with a MW of 111 kDa (incl. a signal sequence) was cloned using degenerate primers. The Bga1 belongs to the Glycosyl hydrolases family 35. Transcriptional analysis of the bga1 expression shows that it is highly expressed on L-arabinose, D-galactose and lower on lactose. Deletion of the gene showed that it is not essential for growth on lactose but that knock-out strains grew slower and produced about half the biomass of the wt strain, whereas amplification of the bga1 gene under the pki1 promoter resulted in faster growth with a reduced lag phase. Bga1 is not necessary for lactose-dependent induction of cbh1 gene expression in H. jecorina but its overexpression impairs lactose-induction of cbh1 expression. Analysis of the deletion strain further showed that bga1 encodes the major extracellular beta-galactosidase which is also partially cell-wall bound. An additional cell wall-bound beta-galactosidase activity could be shown but no evidence for an intracellular beta-galactosidase was obtained.",
author = "Bernhard Seiboth and Lukas Hartl and Noora Salovuori and Jari Vehmaanper{\"a} and Merja Penttil{\"a} and Christian Kubicek",
year = "2003",
doi = "10.4148/1941-4765.1164",
language = "English",
series = "Fungal Genetics Reports",
publisher = "New Prairie Press",
number = "Article 18",
booktitle = "22nd Fungal Genetics Conference at Asilomar",

}

Seiboth, B, Hartl, L, Salovuori, N, Vehmaanperä, J, Penttilä, M & Kubicek, C 2003, Role of the hga1-encoded extracellular beta-galactosidase of Hypocrea jecorina in lactose metabolism and cellulase induction. in 22nd Fungal Genetics Conference at Asilomar: Abstracts from the 2003 Fungal Genetics Conference at Asilomar., 424, Fungal Genetics Reports, no. Article 18, vol. 50, 22nd Fungal Genetics Conference, Asilomar, United States, 18/03/03. https://doi.org/10.4148/1941-4765.1164

Role of the hga1-encoded extracellular beta-galactosidase of Hypocrea jecorina in lactose metabolism and cellulase induction. / Seiboth, Bernhard; Hartl, Lukas; Salovuori, Noora; Vehmaanperä, Jari; Penttilä, Merja; Kubicek, Christian.

22nd Fungal Genetics Conference at Asilomar: Abstracts from the 2003 Fungal Genetics Conference at Asilomar. 2003. 424 (Fungal Genetics Reports; No. Article 18, Vol. 50).

Research output: Chapter in Book/Report/Conference proceedingConference abstract in proceedingsScientific

TY - CHAP

T1 - Role of the hga1-encoded extracellular beta-galactosidase of Hypocrea jecorina in lactose metabolism and cellulase induction

AU - Seiboth, Bernhard

AU - Hartl, Lukas

AU - Salovuori, Noora

AU - Vehmaanperä, Jari

AU - Penttilä, Merja

AU - Kubicek, Christian

PY - 2003

Y1 - 2003

N2 - Lactose is the only economic carbon source for protein production under the control of cellulase promoters by the ascomycete H. jecorina (anamorph: Trichoderma reesei). However, the mechanism by which lactose triggers cellulase formation is not understood. We have investigated the role of beta-galactosidase in lactose metabolism and cellulase induction in H. jecorina. A genomic copy of the bga1 gene predicting a protein with a MW of 111 kDa (incl. a signal sequence) was cloned using degenerate primers. The Bga1 belongs to the Glycosyl hydrolases family 35. Transcriptional analysis of the bga1 expression shows that it is highly expressed on L-arabinose, D-galactose and lower on lactose. Deletion of the gene showed that it is not essential for growth on lactose but that knock-out strains grew slower and produced about half the biomass of the wt strain, whereas amplification of the bga1 gene under the pki1 promoter resulted in faster growth with a reduced lag phase. Bga1 is not necessary for lactose-dependent induction of cbh1 gene expression in H. jecorina but its overexpression impairs lactose-induction of cbh1 expression. Analysis of the deletion strain further showed that bga1 encodes the major extracellular beta-galactosidase which is also partially cell-wall bound. An additional cell wall-bound beta-galactosidase activity could be shown but no evidence for an intracellular beta-galactosidase was obtained.

AB - Lactose is the only economic carbon source for protein production under the control of cellulase promoters by the ascomycete H. jecorina (anamorph: Trichoderma reesei). However, the mechanism by which lactose triggers cellulase formation is not understood. We have investigated the role of beta-galactosidase in lactose metabolism and cellulase induction in H. jecorina. A genomic copy of the bga1 gene predicting a protein with a MW of 111 kDa (incl. a signal sequence) was cloned using degenerate primers. The Bga1 belongs to the Glycosyl hydrolases family 35. Transcriptional analysis of the bga1 expression shows that it is highly expressed on L-arabinose, D-galactose and lower on lactose. Deletion of the gene showed that it is not essential for growth on lactose but that knock-out strains grew slower and produced about half the biomass of the wt strain, whereas amplification of the bga1 gene under the pki1 promoter resulted in faster growth with a reduced lag phase. Bga1 is not necessary for lactose-dependent induction of cbh1 gene expression in H. jecorina but its overexpression impairs lactose-induction of cbh1 expression. Analysis of the deletion strain further showed that bga1 encodes the major extracellular beta-galactosidase which is also partially cell-wall bound. An additional cell wall-bound beta-galactosidase activity could be shown but no evidence for an intracellular beta-galactosidase was obtained.

UR - http://newprairiepress.org/cgi/viewcontent.cgi?article=1164&context=fgr

U2 - 10.4148/1941-4765.1164

DO - 10.4148/1941-4765.1164

M3 - Conference abstract in proceedings

T3 - Fungal Genetics Reports

BT - 22nd Fungal Genetics Conference at Asilomar

ER -

Seiboth B, Hartl L, Salovuori N, Vehmaanperä J, Penttilä M, Kubicek C. Role of the hga1-encoded extracellular beta-galactosidase of Hypocrea jecorina in lactose metabolism and cellulase induction. In 22nd Fungal Genetics Conference at Asilomar: Abstracts from the 2003 Fungal Genetics Conference at Asilomar. 2003. 424. (Fungal Genetics Reports; No. Article 18, Vol. 50). https://doi.org/10.4148/1941-4765.1164