Transformation of Trichoderma reesei with the Hormoconis resinae glucoamylase P (gamP) gene: production of a heterologous glucoamylase by Trichoderma reesei

Joutsjoki, Vesa; Torkkeli, Tuula; Nevalainen, Helena

doi:10.1007/bf00351796

Cited by 39 publications

(17 citation statements)

References 25 publications

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“…The fungal mycelia for DNA isolations were obtained after growing the strains for 2 days on Trichoderma minimal medium containing 2% proteose peptone (Difco). Complex lactose-based cellulase-inducing media (22) were used for enzyme production in shake flasks and fermentations. The transformants were screened using 50-ml cultivations, and the mycelium for the RNA isolations was collected from 200-ml cultivations.…”

Section: Methodsmentioning

confidence: 99%

High-Yield Production of a Bacterial Xylanase in the Filamentous Fungus Trichoderma reesei Requires a Carrier Polypeptide with an Intact Domain Structure

Paloheimo

Mäntylä

Kallio

et al. 2003

Appl Environ Microbiol

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A bacterial xylanase gene, Nonomuraea flexuosa xyn11A, was expressed in the filamentous fungus Trichoderma reesei from the strong cellobiohydrolase 1 promoter as fusions to a variety of carrier polypeptides. By using single-copy isogenic transformants, it was shown that production of this xylanase was clearly increased (up to 820 mg/liter) when it was produced as a fusion protein with a carrier polypeptide having an intact domain structure compared to the production (150 to 300 mg/liter) of fusions to the signal sequence alone or to carriers having incomplete domain structures. The carriers tested were the T. reesei mannanase I (Man5A, or MANI) core-hinge and a fragment thereof and the cellulose binding domain of T. reesei cellobiohydrolase II (Cel6A, or CBHII) with and without the hinge region(s) and a fragment thereof. The flexible hinge region was shown to have a positive effect on both the production of Xyn11A and the efficiency of cleavage of the fusion polypeptide. The recombinant Xyn11A produced had properties similar to those of the native xylanase. It constituted 6 to 10% of the total proteins secreted by the transformants. About three times more of the Man5A core-hinge carrier polypeptide than of the recombinant Xyn11A was observed. Even in the best Xyn11A producers, the levels of the fusion mRNAs were only ϳ10% of the level of cel7A (cbh1) mRNA in the untransformed host strain.

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Section: Methodsmentioning

confidence: 99%

High-Yield Production of a Bacterial Xylanase in the Filamentous Fungus Trichoderma reesei Requires a Carrier Polypeptide with an Intact Domain Structure

Paloheimo

Mäntylä

Kallio

et al. 2003

Appl Environ Microbiol

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“…In T. reesei, it has been shown that the overexpression of the mannosylphosphodolichol synthase-encoding gene from S. cerevisiae, which is required for O-glycan precursor synthesis, increased the production of CBH1 (Kruszewska et al 1999). As for foreign proteins, it has been shown that Candida antarctica lipase produced in A. oryzae (Heogh et al 1995) and Hormoconis resinae glucoamylase P produced in T. reesei (Joutsjoki et al 1993) are overglycosylated. Van den Brink et al (2006) improved a poorly used N-glycosylation site within the prochymosin molecule.…”

Section: Engineering Of the Glycosylation Pathwaymentioning

confidence: 99%

Approaches for refining heterologous protein production in filamentous fungi

Sharma

Katoch

Srivastava

et al. 2009

World J Microbiol Biotechnol

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Fungi combine the advantages of a microbial system such as a simple fermentability with the capability of secreting proteins that are modified according to a general eukaryotic scheme. Filamentous fungi such as Aspergillus niger efficiently secrete genuine proteins but the secretion of recombinant proteins turned out be a difficult task. Aspergillus niger is an attractive organism because of its high secretion capacity and is frequently used as a model organism. Whereas high production yields can be obtained when homologous proteins are expressed, much lower amounts are obtained with the production of heterologous proteins. To fully exploit the potential of filamentous fungi, understanding of the molecular genetics, their physiology, and the glycosylation metabolism has to be investigated and clarified in more detail. This review summarizes recent developments in heterologous protein production by filamentous fungi and also generalizes the possibilities of improving the protein production by various genetic and bioprocessing approaches, thereby easing recognition of filamentous fungi as a relevant and reliable expression platform.

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“…The transformants were inoculated from the PD slants to shake flasks containing 50 ml of complex lactose-based cellulase-inducing medium [37] buffered with 5 % KH 2 PO 4 at pH 6.0. The protease production of the transformants was analyzed from the culture supernatants after growing them for 7 days at 30°C, 250 rpm.…”

Section: Production Of the Recombinant Fe Proteasementioning

confidence: 99%

A New Subtilase-Like Protease Deriving from Fusarium equiseti with High Potential for Industrial Applications

Juntunen

Mäkinen

Isoniemi

et al. 2015

Appl Biochem Biotechnol

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A gene encoding a novel extracellular subtilisin-like protease was cloned from the ascomycete Fusarium equiseti and expressed in Trichoderma reesei. The F. equiseti protease (Fe protease) showed excellent performance in stain removal and good compatibility with several commercial laundry detergent formulations, suggesting that it has high potential for use in various industrial applications. The recombinant enzyme was purified and characterized. The temperature optimum of the Fe protease was 60 °C and it showed high activity in the pH range of 6-10, with a sharp decline in activity at pH above 10. The amino acid specificity of the Fe protease was studied using casein, cytochrome c, and ubiquitin as substrates. The Fe protease had broad substrate specificity: almost all amino acid residues were accepted at position P1, even though it showed some preference for cleavage at the C-terminal side of asparagine and histidine residues. The S4 subsite of Fe protease favors aspartic acid and threonine. The other well-characterized proteases from filamentous fungi, Proteinase K from Engyodontium album, Thermomycolin from Malbranchea sulfurea, and alkaline subtilisins from Bacillus species prefer hydrophobic amino acids in both the S1 and S4 subsites. Due to its different specificity compared to the members of the S8 family of clan SB of proteases, we consider that the Fe protease is a new protease. It does not belong to any previously defined IUBMB groups of proteases.

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Transformation of Trichoderma reesei with the Hormoconis resinae glucoamylase P (gamP) gene: production of a heterologous glucoamylase by Trichoderma reesei

Cited by 39 publications

References 25 publications

High-Yield Production of a Bacterial Xylanase in the Filamentous Fungus Trichoderma reesei Requires a Carrier Polypeptide with an Intact Domain Structure

High-Yield Production of a Bacterial Xylanase in the Filamentous Fungus Trichoderma reesei Requires a Carrier Polypeptide with an Intact Domain Structure

Approaches for refining heterologous protein production in filamentous fungi

A New Subtilase-Like Protease Deriving from Fusarium equiseti with High Potential for Industrial Applications

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