Lipid production by robust Aspergillus oryzae BCC7051 and a mathematical model describing its growth and lipid phenotypic traits.

Aims: To identify the promising oleaginous Aspergillus oryzae strain and leverage its lipid and biomass production through a mathematical model.

Methods And Results: Comparative profiling of the cell growth and total fatty acid (TFA) content among 13 strains of A. oryzae was performed to explore the discrimination in their lipid productions.

Holistic transcriptional responses of Cordyceps militaris to different culture temperatures.

Cordyceps militaris is a medicinal entomopathogenic fungus containing valuable biometabolites for pharmaceutical applications. Its genetic inheritance and environmental factors play a crucial role in the production of biomass enriched with cordycepin. While temperature is a crucial controlled parameter for fungal cultivation, its impacts on growth and metabolite biosynthesis remains poorly characterized.

Dissecting Holistic Metabolic Acclimatization of WJ11 Defective in Carotenoid Biosynthesis.

WJ11 is a lipid-producing strain with industrial potential. A holistic approach using gene manipulation and bioprocessing development has improved lipid production and the strain's economic viability. However, the systematic regulation of lipid accumulation and carotenoid biosynthesis in remains unknown.

Light-Exposed Metabolic Responses of through Transcriptome-Integrated Genome-Scale Modeling.

The genome-scale metabolic model (GSMM) of provides a comprehensive basis of carbon assimilation for cell growth and metabolite production. However, the model with a simple mass balance concept shows limited capability to probe the metabolic responses of under light exposure. This study, therefore, employed the transcriptome-integrated GSMM approach to extend the investigation of 's metabolism under light conditions.

Efficient de novo production of bioactive cordycepin by Aspergillus oryzae using a food-grade expression platform.

Background: Cordycepin (3'-deoxyadenosine) is an important bioactive compound in medical and healthcare markets. The drawbacks of commercial cordycepin production using Cordyceps spp. include long cultivation periods and low cordycepin yields.

Functional genomics and systems biology of Cordyceps species for biotechnological applications.

The use of Cordyceps species for the manufacture of natural products has been established; however, the tremendous advances observed in recent years in genetic engineering and molecular biology have revolutionized the optimization of Cordyceps as cell factories and drastically expanded the biotechnological potential of these fungi. Here, we present a review of systems and synthetic biology studies of Cordyceps and their implications for fungal biology and industrial applications. We summarize the current status of synthetic biology for enhancing targeted metabolites in Cordyceps species, such as cordycepin, adenosine, polysaccharide, and pentostatin.

Transcriptome-based Mining of the Constitutive Promoters for Tuning Gene Expression in Aspergillus oryzae.

Transcriptional regulation has been adopted for developing metabolic engineering tools. The regulatory promoter is a crucial genetic element for strain optimization. In this study, a gene set of Aspergillus oryzae with highly constitutive expression across different growth stages was identified through transcriptome data analysis.

Enhancing Genome-Scale Model by Integrative Exometabolome and Transcriptome: Unveiling Carbon Assimilation towards Sphingolipid Biosynthetic Capability of .

is an industrially important fungus, which is often used in Asia as traditional medicine. There has been a published genome-scale metabolic model (GSMM) of useful for predicting its growth behaviors; however, lipid metabolism, which plays a vital role in cellular functions, remains incomplete in the GSMM of . A comprehensive study on was thus performed by enhancing GSMM through integrative analysis of metabolic footprint and transcriptome data.

The Exploring Functional Role of Ammonium Transporters of in Nitrogen Metabolism: Challenges towards Cell Biomass Production.

Ammonium is a source of fermentable inorganic nitrogen essential for the growth and development of filamentous fungi. It is involved in several cellular metabolic pathways underlying nitrogen transport and assimilation. Ammonium can be transferred into the cell by an ammonium transporter.

Potential of Aspergillus oryzae as a biosynthetic platform for indigoidine, a non-ribosomal peptide pigment with antioxidant activity.

The growing demand for natural pigments in the industrial sector is a significant driving force in the development of production processes. The production of natural blue pigments, which have wide industrial applications, using microbial systems has been gaining significant attention. In this study, we used Aspergillus oryzae as a platform cell factory to produce the blue pigment indigoidine (InK), by genetic manipulation of its non-ribosomal peptide synthetase system to overexpress the indigoidine synthetase gene (AoinK).

Novel pentose-regulated promoter of with application in controlling heterologous gene expression.

The potent promoter and its transcriptional control make a significant contribution to strain optimization. Using transcriptomebased approach, a novel pentose-regulated promoter of the xylose reductase gene () of  was identified. The promoter analysis showed that the  was tightly regulated by pentose sugars, which xylose and xylan were favorable inducers.

Promoter exchange of the cryptic nonribosomal peptide synthetase gene for oligopeptide production in Aspergillus oryzae.

Oligopeptides with functional activities are of current interest in the nutraceutical and medical sectors. The development of the biosynthetic process of oligopeptides through a nonribosomal peptide synthetase (NRPS) system has become more challenging. To develop a production platform for nonribosomal peptides (NRPs), reprogramming of transcriptional regulation of the acv gene encoded ACV synthetase (ACVS) was implemented in Aspergillus oryzae using the CRISPR-Cas9 system.

Metabolic Regulation of Sugar Assimilation for Lipid Production in BCC7051 through Comparative Transcriptome Perspective.

Microbial lipid production with cost effectiveness is a prerequisite for the oleochemical sector. In this work, genome-wide transcriptional responses on the utilization of xylose and glucose in oleaginous were studied with relation to growth and lipid phenotypic traits. Comparative analysis of the active growth (t1) and lipid-accumulating (t2) stages showed that the C5 cultures efficiently consumed carbon sources for biomass and lipid production comparable to the C6 cultures.

Significance of two intracellular triacylglycerol lipases of Aspergillus oryzae in lipid mobilization: A perspective in industrial implication for microbial lipid production.

Microbial lipid production of oleaginous strains involves in a complex cellular metabolism controlling lipid biosynthesis, accumulation and degradation. Particular storage lipid, triacylglycerol (TAG), contributes to dynamic traits of intracellular lipids and cell growth. To explore a basis of TAG degradation in the oleaginous strain of Aspergillus oryzae, the functional role of two intracellular triacylglycerol lipases, AoTgla and AoTglb, were investigated by targeted gene disruption using CRISPR/Cas9 system.

Physiological Traits of Dihomo-γ-Linolenic Acid Production of the Engineered by Comparing Mathematical Models.

Dihomo-γ-linolenic acid (DGLA; C20:3 -6) is expected to dominate the functional ingredients market for its role in anti-inflammation and anti-proliferation. The DGLA production by the engineered strain of with overexpressing Δ-desaturase and Δ-elongase genes was investigated by manipulating the nutrient and fermentation regimes. Of the nitrogen sources tested, the maximum biomass and DGLA titers were obtained in the cultures using NaNO grown at pH 6.

Exploring differential traits of lipid-producing stages of the wild type and morphologically engineered strain of Aspergillus oryzae by comparative kinetic modeling.

Comparative profilings of cell growth and lipid production in the morphologically engineered strain (Δags1) and the wild type (WT) of Aspergillus oryzae BCC7051 were implemented. Using various nitrogen sources, a discrimination in cell morphology between the two strains was found, of which the Δags1 culture exhibited mycelial growth as small pellets in contrast to the WT. Of them, sodium nitrate and potassium nitrate were optimal for lipid production of the WT and Δags1 strains, respectively, which the highest lipid concentrations of 7.

Metabolic Responses of Carotenoid and Cordycepin Biosynthetic Pathways in under Light-Programming Exposure through Genome-Wide Transcriptional Analysis.

is currently exploited for commercial production of specialty products as its biomass constituents are enriched in bioactive compounds, such as cordycepin. The rational process development is important for economically feasible production of high quality bioproducts. Light is an abiotic factor affecting the cultivation process of this entomopathogenic fungus, particularly in its carotenoid formation.

Systematic genome analysis of a novel arachidonic acid-producing strain uncovered unique metabolic traits in the production of acetyl-CoA-derived products in Mortierellale fungi.

The fungi in order Mortierellales are attractive producers for long-chain polyunsaturated fatty acids (PUFAs). Here, the genome sequencing and assembly of a novel strain of Mortierella sp. BCC40632 were done, yielding 65 contigs spanning of 49,964,116 total bases with predicted 12,149 protein-coding genes.

Probing Carbon Utilization of by Sugar Transportome and Protein Structural Analysis.

Beyond comparative genomics, we identified 85 sugar transporter genes in , clustering into nine subfamilies as sequence- and phylogenetic-based functional classification, presuming the versatile capability of the fungal growths on a range of sugars. Further analysis of the global gene expression patterns of showed 123 genes were significantly expressed across the sucrose, glucose, and xylose cultures. The sugar transporters specific for pentose were then identified by gene-set enrichment analysis.

Functional Characterization of Novel U6 RNA Polymerase III Promoters: Their Implication for CRISPR-Cas9-Mediated Gene Editing in Aspergillus oryzae.

U6 RNA polymerase III promoter (PU6), which is a key element in controlling the generation of single-guide RNA (sgRNA) for gene editing through CRISPR-Cas9 system, was investigated in this work. Using bioinformatics approach, two novel U6 ribonucleic acid (U6 RNA) sequences of Aspergillus niger were identified, showing that they had conserved motifs similar to other U6 RNAs. The putative PU6 located at the upstream sequence of A.

Morphologically engineered strain of Aspergillus oryzae as a cell chassis for production development of functional lipids.

Cell morphology of the oleaginous fungus, Aspergillus oryzae BCC7051, was genetically engineered by disruption of non-essential genes involved in cell wall biosynthesis. Comparative phenotypic analysis of two disruptant strains defective either in α-1,3-glucan synthase 1 (ΔAoAgs1) or chitin synthase B (ΔAoChsB), and the wild type showed that the ΔAoAgs1 strain had no alterations in colonial growth and sporulation when grown on agar medium whereas the ΔAoChsB disruptant showed growth retardation and lower sporulation. However, tiny and loose pellets were found in the ΔAoAgs1 culture grown in liquid medium, where fungal pellet size was decreased by 35-50% of the wild type size.

Metabolic traits specific for lipid-overproducing strain of WJ11 identified by genome-scale modeling approach.

The genome-scale metabolic model of a lipid-overproducing strain of WJ11 was developed. The model (NI1159) contained 1,159 genes, 648 EC numbers, 1,537 metabolites, and 1,355 metabolic reactions, which were localized in different compartments of the cell. Using flux balance analysis (FBA), the NI1159 model was validated by predicting the specific growth rate.

Reengineering lipid biosynthetic pathways of Aspergillus oryzae for enhanced production of γ-linolenic acid and dihomo-γ-linolenic acid.

Biological significance of 18-carbon polyunsaturated fatty acids, γ-linolenic acid (GLA; C18:3 n-6) and dihomo-γ-linolenic acid (DGLA; C20:3 n-6) has gained much attention in the systematic development of optimized strains for industrial applications. In this work, a n-6 PUFAs-producing strain of Aspergillus oryzae was generated by manipulating metabolic reactions in fatty acid modification and triacylglycerol biosynthesis. The codon-optimized genes coding for Δ-desaturase and Δ-elongase of Pythium sp.

Alternative metabolic routes in channeling xylose to cordycepin production of Cordyceps militaris identified by comparative transcriptome analysis.

The responsive mechanism of C. militaris TBRC7358 on xylose utilization was investigated by comparative analysis of transcriptomes, growth kinetics and cordycepin productions. The result showed that the culture grown on xylose exhibited high production yield of cordycepin on dry biomass.

Uncovering global metabolic response to cordycepin production in Cordyceps militaris through transcriptome and genome-scale network-driven analysis.

The cellular metabolic adaptations of Cordyceps militaris have been progressively studied. In particular, the cordycepin pathway is of interest in medicinal applications. Even though the metabolic pathways for cordycepin production are known to be related to different carbon sources, the regulatory mechanisms at a systems level are poorly characterized.