Genetic modification of carbon catabolite repression in Trichoderma reesei for improved protein production

Tiina Nakari-Setälä, Marja Paloheimo, Jarno Kallio, Jari Vehmaanperä, Merja Penttilä, Markku Saloheimo

Research output: Contribution to journalArticleScientificpeer-review

102 Citations (Scopus)

Abstract

The cellulase and hemicellulase genes of the filamentous fungus Trichoderma reesei have been shown to be under carbon catabolite repression mediated by the regulatory gene cre1. In this study, strains were constructed in which the cre1 gene was either completely removed or replaced by a truncated mutant variant, cre1-1, found previously in the Rut-C30 mutant strain with enhanced enzyme production capability. The T. reesei transformants with either deletion or truncation of cre1 had clearly altered colony morphology compared with the parental strains, forming smaller colonies and fewer aerial hyphae and spores. Liquid cultures in a medium with glucose as a carbon source showed that the transformants were derepressed in cellulase and hemicellulase production. Interestingly, they also produced significantly elevated levels of these hydrolytic enzymes in fermentations carried out in a medium inducing the hydrolase genes. This suggests that cre1 acts as a modulator of cellulase and hemicellulase gene expression under both noninducing and inducing conditions. There was no phenotypic difference between the Δcre1 and cre1-1 mutant strains in any of the experiments done, indicating that the cre1-1 gene is practically a null allele. The results of this work indicate that cre1 is a valid target gene in strain engineering for improved enzyme production in T. reesei.

Original languageEnglish
Pages (from-to)4853-4860
Number of pages8
JournalApplied and Environmental Microbiology
Volume75
Issue number14
DOIs
Publication statusPublished - 1 Jul 2009
MoE publication typeA1 Journal article-refereed

Fingerprint

Catabolite Repression
Trichoderma reesei
Trichoderma
genetic engineering
Cellulase
metabolites
protein
endo-1,4-beta-glucanase
gene
carbon
Genes
Proteins
genes
proteins
mutants
enzyme
Enzymes
enzymes
null alleles
Hyphae

Keywords

  • Trichoderma reesei
  • catabolite inactivation
  • protein production

Cite this

@article{26acf18dd3034f8c8ca30aba34bb2904,
title = "Genetic modification of carbon catabolite repression in Trichoderma reesei for improved protein production",
abstract = "The cellulase and hemicellulase genes of the filamentous fungus Trichoderma reesei have been shown to be under carbon catabolite repression mediated by the regulatory gene cre1. In this study, strains were constructed in which the cre1 gene was either completely removed or replaced by a truncated mutant variant, cre1-1, found previously in the Rut-C30 mutant strain with enhanced enzyme production capability. The T. reesei transformants with either deletion or truncation of cre1 had clearly altered colony morphology compared with the parental strains, forming smaller colonies and fewer aerial hyphae and spores. Liquid cultures in a medium with glucose as a carbon source showed that the transformants were derepressed in cellulase and hemicellulase production. Interestingly, they also produced significantly elevated levels of these hydrolytic enzymes in fermentations carried out in a medium inducing the hydrolase genes. This suggests that cre1 acts as a modulator of cellulase and hemicellulase gene expression under both noninducing and inducing conditions. There was no phenotypic difference between the Δcre1 and cre1-1 mutant strains in any of the experiments done, indicating that the cre1-1 gene is practically a null allele. The results of this work indicate that cre1 is a valid target gene in strain engineering for improved enzyme production in T. reesei.",
keywords = "Trichoderma reesei, catabolite inactivation, protein production",
author = "Tiina Nakari-Set{\"a}l{\"a} and Marja Paloheimo and Jarno Kallio and Jari Vehmaanper{\"a} and Merja Penttil{\"a} and Markku Saloheimo",
year = "2009",
month = "7",
day = "1",
doi = "10.1128/AEM.00282-09",
language = "English",
volume = "75",
pages = "4853--4860",
journal = "Applied and Environmental Microbiology",
issn = "0099-2240",
publisher = "American Society for Microbiology",
number = "14",

}

Genetic modification of carbon catabolite repression in Trichoderma reesei for improved protein production. / Nakari-Setälä, Tiina; Paloheimo, Marja; Kallio, Jarno; Vehmaanperä, Jari; Penttilä, Merja; Saloheimo, Markku.

In: Applied and Environmental Microbiology, Vol. 75, No. 14, 01.07.2009, p. 4853-4860.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Genetic modification of carbon catabolite repression in Trichoderma reesei for improved protein production

AU - Nakari-Setälä, Tiina

AU - Paloheimo, Marja

AU - Kallio, Jarno

AU - Vehmaanperä, Jari

AU - Penttilä, Merja

AU - Saloheimo, Markku

PY - 2009/7/1

Y1 - 2009/7/1

N2 - The cellulase and hemicellulase genes of the filamentous fungus Trichoderma reesei have been shown to be under carbon catabolite repression mediated by the regulatory gene cre1. In this study, strains were constructed in which the cre1 gene was either completely removed or replaced by a truncated mutant variant, cre1-1, found previously in the Rut-C30 mutant strain with enhanced enzyme production capability. The T. reesei transformants with either deletion or truncation of cre1 had clearly altered colony morphology compared with the parental strains, forming smaller colonies and fewer aerial hyphae and spores. Liquid cultures in a medium with glucose as a carbon source showed that the transformants were derepressed in cellulase and hemicellulase production. Interestingly, they also produced significantly elevated levels of these hydrolytic enzymes in fermentations carried out in a medium inducing the hydrolase genes. This suggests that cre1 acts as a modulator of cellulase and hemicellulase gene expression under both noninducing and inducing conditions. There was no phenotypic difference between the Δcre1 and cre1-1 mutant strains in any of the experiments done, indicating that the cre1-1 gene is practically a null allele. The results of this work indicate that cre1 is a valid target gene in strain engineering for improved enzyme production in T. reesei.

AB - The cellulase and hemicellulase genes of the filamentous fungus Trichoderma reesei have been shown to be under carbon catabolite repression mediated by the regulatory gene cre1. In this study, strains were constructed in which the cre1 gene was either completely removed or replaced by a truncated mutant variant, cre1-1, found previously in the Rut-C30 mutant strain with enhanced enzyme production capability. The T. reesei transformants with either deletion or truncation of cre1 had clearly altered colony morphology compared with the parental strains, forming smaller colonies and fewer aerial hyphae and spores. Liquid cultures in a medium with glucose as a carbon source showed that the transformants were derepressed in cellulase and hemicellulase production. Interestingly, they also produced significantly elevated levels of these hydrolytic enzymes in fermentations carried out in a medium inducing the hydrolase genes. This suggests that cre1 acts as a modulator of cellulase and hemicellulase gene expression under both noninducing and inducing conditions. There was no phenotypic difference between the Δcre1 and cre1-1 mutant strains in any of the experiments done, indicating that the cre1-1 gene is practically a null allele. The results of this work indicate that cre1 is a valid target gene in strain engineering for improved enzyme production in T. reesei.

KW - Trichoderma reesei

KW - catabolite inactivation

KW - protein production

UR - http://www.scopus.com/inward/record.url?scp=67650469491&partnerID=8YFLogxK

U2 - 10.1128/AEM.00282-09

DO - 10.1128/AEM.00282-09

M3 - Article

VL - 75

SP - 4853

EP - 4860

JO - Applied and Environmental Microbiology

JF - Applied and Environmental Microbiology

SN - 0099-2240

IS - 14

ER -