Optimization and stability of glucoamylase production by recombinant strains ofAspergillus niger in chemostat culture

Withers, Julie M.; Swift, Richard J.; Wiebe, Marilyn G.; Robson, Geoffrey D.; Punt, Peter J.; Hondel, Cees A. M. J. J. van den; Trinci, Anthony P. J.

doi:10.1002/(sici)1097-0290(19980820)59:4<407::aid-bit3>3.0.co;2-k

Cited by 64 publications

(7 citation statements)

References 36 publications

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“…In contrast to most processes with yeast and bacteria, protein expression in Aspergillus niger has often been carried out using chromosomal expression instead of plasmid-based systems [ 53 , 54 , 55 , 56 ]. Often, hosts with multiple copies of the target gene are used for stronger expression [ 53 , 55 ]. In most cases, the product formation kinetics seem to be growth coupled and highest q p have been reached at µ > 0.1 h −1 [ 53 , 54 , 55 , 56 ].…”

Section: μ-Dependent Recombinant Product Formation Kineticsmentioning

confidence: 99%

Application of Continuous Culture Methods to Recombinant Protein Production in Microorganisms

Peebo

Neubauer

2018

Microorganisms

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Depending on the environmental conditions, cells adapt their metabolism and specific growth rate. Rearrangements occur on many different levels such as macromolecular composition, gene and protein expression, morphology and metabolic flux patterns. As the interplay of these processes also determines the output of a recombinant protein producing system, having control over specific growth rate of the culture is advantageous. Continuous culture methods were developed to grow cells in a constant environment and have been used for decades to study basic microbial physiology in a controlled and reproducible manner. Our review summarizes the uses of continuous cultures in cell physiology studies and process development, with a focus on recombinant protein-producing microorganisms.

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Section: μ-Dependent Recombinant Product Formation Kineticsmentioning

confidence: 99%

Application of Continuous Culture Methods to Recombinant Protein Production in Microorganisms

Peebo

Neubauer

2018

Microorganisms

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“…Chemostats, in which fresh medium is continually added to a bioreactor, and effluent containing metabolite products, used medium, and microbial biomass continually removed, enables steady state microbial growth. Accordingly, optimal growth rates for product biosynthesis can be maintained, and the period of product biosynthesis increased when compared to batch cultivation [8]. …”

Section: Introductionmentioning

confidence: 99%

Highly active promoters and native secretion signals for protein production during extremely low growth rates in Aspergillus niger

Wanka¹,

Arentshorst

Cairns³

et al. 2016

Microb Cell Fact

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BackgroundThe filamentous ascomycete Aspergillus niger is used in many industrial processes for the production of enzymes and organic acids by batch and fed-batch cultivation. An alternative technique is continuous cultivation, which promises improved yield and optimized pipeline efficiency.ResultsIn this work, we have used perfusion (retentostat) cultivation to validate two promoters that are suitable for A. niger continuous cultivation of industrially relevant products. Firstly, promoters of genes encoding either an antifungal protein (Panafp) or putative hydrophobin (PhfbD) were confirmed as active throughout retentostat culture by assessing mRNA and protein levels using a luciferase (mluc) reporter system. This demonstrated the anafp promoter mediates a high but temporally variable expression profile, whereas the hfbD promoter mediates a semi-constant, moderate-to-high protein expression during retentostat culture. In order to assess whether these promoters were suitable to produce heterologous proteins during retentostat cultivation, the secreted antifungal protein (AFP) from Aspergillus giganteus, which has many potential biotechnological applications, was expressed in A. niger during retentostat cultivation. Additionally, this assay was used to concomitantly validate that native secretion signals encoded in anafp and hfbD genes can be harnessed for secretion of heterologous proteins. Afp mRNA and protein abundance were comparable to luciferase measurements throughout retentostat cultivation, validating the use of Panafp and PhfbD for perfusion cultivation. Finally, a gene encoding the highly commercially relevant thermal hysteresis protein (THP) was expressed in this system, which did not yield detectable protein.ConclusionBoth hfbD and anafp promoters are suitable for production of useful products in A. niger during perfusion cultivation. These findings provide a platform for further optimisations for high production of heterologous proteins with industrial relevance.Electronic supplementary materialThe online version of this article (doi:10.1186/s12934-016-0543-2) contains supplementary material, which is available to authorized users.

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“…Production of extracellular proteins by filamentous fungi, such as α-amylase in Aspergillus oryzae [2], [17] and glucoamylase in Aspergillus niger [11], [16], [22], has been shown to be growth-associated in many cases. However, examples of non growth associated production are also known, such as recombinant protein production by Fusarium venenatum [21].…”

Section: Discussionmentioning

confidence: 99%

High cell density cultivation of six fungal strains efficient in azo dye bioremediation

El‐Rahim

Mostafa

Moawad

2016

Biotechnology Reports

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HighlightsFungal strains having the potential of high cell density accumulation to be used for textile azo dyes bioremediation.The optimization of growth conditions for enhancing the biomass accumulation and protein production by six fungal strains was studied in 7 liter fermentor.The maximum biomass accumulation and protein production were achieved in molasses containing.The molasses as a cheaper carbon source was successfully used to promote fungal biomass and protein accumulation by the azo dye removing fungal strains.

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Optimization and stability of glucoamylase production by recombinant strains ofAspergillus niger in chemostat culture

Cited by 64 publications

References 36 publications

Application of Continuous Culture Methods to Recombinant Protein Production in Microorganisms

Application of Continuous Culture Methods to Recombinant Protein Production in Microorganisms

Highly active promoters and native secretion signals for protein production during extremely low growth rates in Aspergillus niger

High cell density cultivation of six fungal strains efficient in azo dye bioremediation

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