New Promoters to Improve Heterologous Protein Production in Aspergillus vadensis

Appl Microbiol Biotechnol

2023

Filamentous fungi are able to produce a wide range of valuable proteins and enzymes for many industrial applications. Recent advances in fungal genomics and experimental technologies are rapidly changing the approaches for the development and use of filamentous fungi as hosts for the production of both homologous and heterologous proteins. In this review, we highlight the benefits and challenges of using filamentous fungi for the production of heterologous proteins. We review various techniques commonly employed to improve the heterologous protein production in filamentous fungi, such as strong and inducible promoters, codon optimization, more efficient signal peptides for secretion, carrier proteins, engineering of glycosylation sites, regulation of the unfolded protein response and endoplasmic reticulum associated protein degradation, optimization of the intracellular transport process, regulation of unconventional protein secretion, and construction of protease-deficient strains. Key points • This review updates the knowledge on heterologous protein production in filamentous fungi. • Several fungal cell factories and potential candidates are discussed. • Insights into improving heterologous gene expression are given.

Section: Main Fungi Used As Protein Cell Factoriesmentioning

confidence: 99%

Heterologous protein production in filamentous fungi

Liu

Garrigues

Appl Microbiol Biotechnol

2023

“…As described by Lagopodi et al [24], the pGPDGFP vector is composed of the gfp gene under the control of the gpdA promoter from A. nidulans [31] and terminated with the trpC terminator [28]. A new expression vector was built based on the pGP-DGFP vector where pgpdA from A. nidulans was replaced with pgpdA from A. vadensis [10] using restrictions enzymes NotI/NcoI. Due to the presence of an internal HindIII within pgpdA from A. vadensis, the HindIII site at the 3′ end of the gfp gene was replaced with AflII and a 6× Histidine tag and stop codon were added to assist in the downstream purification of the enzymes once expressed.…”

Section: Pcr and Expression Vectorsmentioning

confidence: 99%

“…For expression and characterization of the selected genes, the corresponding fragments were cloned behind the gpdA promoter from A. vadensis [10]. The resulting expression vectors containing the gene sequence were then transformed into A. vadensis CBS113365.…”

Section: Development Of Expression Strainsmentioning

confidence: 99%

Overexpression, purification and characterisation of homologous α-l-arabinofuranosidase and endo-1,4-β-d-glucanase in Aspergillus vadensis

Culleton

McKie²,

Journal of Industrial Microbiology and Biotechnology

2014

Self Cite

demonstrated interesting differences in temperate optima, pH stability and substrate specificities. The endo-1,4-β-dglucanase from A. vadensis exhibited a pH and temperature optimum of pH 4.5 and 50 °C, respectively. Comparative biochemical analysis to the orthologous EglA from A. niger presented similar pH and substrate specificity profiles. However, significant differences in temperature optima and stability were noted.

“…However, the application of mutagenesis to obtain hosts with low background expression is labor intensive. In addition to this, naturally occurring low-protease-producing fungal strains, such as Aspergillus vadensis, have been reported as alternative fungal strains for heterologous protein production [ 15 ]. In any case, filamentous fungi have undoubtedly become major contributors to the production of the majority of microbial-derived industrial products [ 16 ] and have become essential contributors to the circular bio-economy [ 2 ].…”

Section: Introductionmentioning

confidence: 99%

Strategies for the Development of Industrial Fungal Producing Strains

Salazar-Cerezo

Garrigues

2023

JoF

The use of microorganisms in industry has enabled the (over)production of various compounds (e.g., primary and secondary metabolites, proteins and enzymes) that are relevant for the production of antibiotics, food, beverages, cosmetics, chemicals and biofuels, among others. Industrial strains are commonly obtained by conventional (non-GMO) strain improvement strategies and random screening and selection. However, recombinant DNA technology has made it possible to improve microbial strains by adding, deleting or modifying specific genes. Techniques such as genetic engineering and genome editing are contributing to the development of industrial production strains. Nevertheless, there is still significant room for further strain improvement. In this review, we will focus on classical and recent methods, tools and technologies used for the development of fungal production strains with the potential to be applied at an industrial scale. Additionally, the use of functional genomics, transcriptomics, proteomics and metabolomics together with the implementation of genetic manipulation techniques and expression tools will be discussed.