Secretion of the sweet-tasting protein thaumatin by recombinant strains of Aspergillus niger var. awamori

Faus, Ignacio; Moral, Catalina del; Adroer, Núria; Río, José Luis del; Patiño, Cristina; Sisniega, Heidi; Casas, C.; Bladé, J.; Rubio, Víctor J.

doi:10.1007/s002530051188

Cited by 31 publications

(16 citation statements)

References 14 publications

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“…Another expression system using A. niger var. awamori has been tried with secretion of thaumatin in concentrations of 5 -7 mgL À1 (Faus et al, 1998).…”

Section: Introductionmentioning

confidence: 99%

Cloning, expression and characterization of recombinant sweet‐protein thaumatin II using the methylotrophic yeast Pichia pastoris

Masuda

Tamaki

Kaneko

et al. 2004

Biotech & Bioengineering

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Thaumatin, an intensely sweet-tasting protein, was secreted by the methylotrophic yeast Pichia pastoris. The mature thaumatin II gene was directly cloned from Taq polymerase-amplified PCR products by using TA cloning methods and fused the pPIC9K expression vector that contains Saccharomyces cerevisiae prepro alpha-mating factor secretion signal. Several additional amino acid residues were introduced at both the N- and C-terminal ends by genetic modification to investigate the role of the terminal end region for elicitation of sweetness in the thaumatin molecule. The secondary and tertiary structures of purified recombinant thaumatin were almost identical to those of the plant thaumatin molecule. Recombinant thaumatin II elicited a sweet taste as native plant thaumatin II; its threshold value of sweetness to humans was around 50 nM, which is the same as that of plant thaumatin II. These results demonstrate that the functional expression of thaumatin II was attained by Pichia pastoris systems and that the N- and C-terminal regions of the thaumatin II molecule do not -play an important role in eliciting the sweet taste of thaumatin.

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“…Another expression system using A. niger var. awamori has been tried with secretion of thaumatin in concentrations of 5 -7 mgL À1 (Faus et al, 1998).…”

Section: Introductionmentioning

confidence: 99%

Cloning, expression and characterization of recombinant sweet‐protein thaumatin II using the methylotrophic yeast Pichia pastoris

Masuda

Tamaki

Kaneko

et al. 2004

Biotech & Bioengineering

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“…Strong induction of glaA-promoter controlled gene expression by maltose has been described before for glucoamylase production (Barton et al, 1972;Chiquetto et al, 1992;Fowler et al, 1990) as well as for glaA-promoter controlled production of heterologous proteins (Faus et al, 1998;Verdoes et al, 1994b;Ward et al, 1997;Wiebe et al, 2001). Maltotriose induced equally high levels of GFP fluorescence as maltose.…”

Section: High Throughput Screening Of Different Metabolizable Carbon mentioning

confidence: 69%

In-depth analysis of the Aspergillus niger glucoamylase (glaA) promoter performance using high-throughput screening and controlled bioreactor cultivation techniques

Ganzlin

Rinas

2008

Journal of Biotechnology

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CitationIn-depth analysis of the Aspergillus niger glucoamylase (glaA) promoter performance using high-throughput screening and controlled bioreactor cultivation techniques. However, induction of the glaA expression system through the monosaccharide glucose is significantly lower compared to starch and the higher molecular weight starch degradation products. All other 26 carbon substrates tested do not induce, or even, as in the case of the easily metabolizable monosaccharide xylose, repress glaA-promoter controlled gene expression in the presence of the inducing disaccharide maltose with an increase of repression strength by increasing xylose concentrations. The complex effect of glucose on glaA-promoter controlled expression was also analyzed using non-metabolizable glucose analogs, namely 5-thio-glucose and 2-deoxyglucose, which were identified as novel and potent inducers of the glaA expression system. The results show that the induction strength depends on the inducer concentration with a maximum at defined concentrations and lower induction or even repression at concentrations above. Moreover, controlled fed-batch cultivations using a high maltose feed rate with concomitant extracellular accumulation of glucose resulted in lower levels of the reporter protein compared to cultures with a low maltose feed rate without extracellular glucose accumulation, thus supporting the conclusion that increasing the glucose concentration beyond a critical point reduces the induction strength or may even cause repression. This way, the speed of polymer hydrolysis, glucose uptake and intracellular breakdown can be fine-tuned for optimal fungal growth and the metabolic burden for glucoamylase synthesis can be limited adequately in response to nutrient availability.-3 -

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“…There has also been considerable work towards cloning of Aspergillus and other fungal genes and their expression in other organisms, including plants; for example fungal phytases have been expressed in crop plants in order to improve yields by enhancing phosphorous uptake from soil, while increasing the nutritive value for animals and reducing water eutrophication from agricultural runoV [1][2][3][4]. While most of the biologically active metabolites developed as pharmaceuticals are derived from plants, as yet there has been little work on expressing plant genes in Aspergillus; the major examples to date being the sweetening proteins thaumatin and curculin (neoculin) [5,6]. Recombinantly-produced human proteins and antibody products are emerging as an important new sector in therapeutics, however historically most drugs have been derived from small molecules, such as secondary metabolites.…”

Section: Introductionmentioning

confidence: 99%

Expression of a synthetic Artemesia annua amorphadiene synthase in Aspergillus nidulans yields altered product distribution

Lubertozzi

Keasling

2008

J Ind Microbiol Biotechnol

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A gene encoding a plant terpene cyclase, Artemisia annua amorpha-4,11-diene synthase (ADS), was expressed in Aspergillus nidulans under control of a strong constitutive promoter, (p)gpdA. The transformants produced only small amounts of amorphadiene, but much larger amounts of similar sesquiterpenes normally produced as minor by-products in planta. In contrast, expression of ADS in Escherichia coli produced almost exclusively amorpha-4,11-diene. These results indicate that the host environment can greatly impact the terpenes produced from terpene synthases.

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Secretion of the sweet-tasting protein thaumatin by recombinant strains of Aspergillus niger var. awamori

Cited by 31 publications

References 14 publications

Cloning, expression and characterization of recombinant sweet‐protein thaumatin II using the methylotrophic yeast Pichia pastoris

Cloning, expression and characterization of recombinant sweet‐protein thaumatin II using the methylotrophic yeast Pichia pastoris

In-depth analysis of the Aspergillus niger glucoamylase (glaA) promoter performance using high-throughput screening and controlled bioreactor cultivation techniques

Expression of a synthetic Artemesia annua amorphadiene synthase in Aspergillus nidulans yields altered product distribution

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