Biotechnological production of fuels, chemicals and proteins is dependent on efficient systems, typically genetically engineered microorganisms. New genome editing methods are making it increasingly easy to introduce new genes functionalities in a broad range organisms. However, engineering all these organisms hampered by the lack suitable gene expression tools. Here, we describe synthetic system (SES) that functional spectrum fungal species without need for host-dependent optimization. The...

Abstract Trichoderma reesei is an established protein production host with high natural capacity to secrete enzymes. The lack of efficient genome engineering approaches and absence robust constitutive gene expression systems limits exploitation this organism in some applications. Here we report T . for high-level highly enriched lipase B Candida antarctica (calB) using glucose as a carbon source. Multiplexed CRISPR/Cas9 combination the use our recently synthetic system (SES) enabled...

The inositolphosphosphingolipid phospholipase C (Isc1p) of Saccharomyces cerevisiae belongs to the family neutral sphingomyelinases that generates bioactive sphingolipid ceramide. In this work role Isc1p in oxidative stress resistance and chronological lifespan was investigated. Loss resulted a higher sensitivity hydrogen peroxide associated with an increase markers, namely intracellular oxidation, protein carbonylation, lipid peroxidation. Microarray analysis showed deficiency up-regulated...

Sustainable production of chemicals, materials, and pharmaceuticals is increasingly performed by genetically engineered cell factories. Engineering complex metabolic routes or behavior control systems requires robust predictable gene expression tools. In this challenging task, orthogonality a fundamental prerequisite for such study, we developed characterized in depth comprehensive toolkit that allows accurate Saccharomyces cerevisiae without marked interference with native cellular...

ABSTRACT d -Galacturonic acid can be obtained by hydrolyzing pectin, which is an abundant and low value raw material. By means of metabolic engineering, we constructed fungal strains for the conversion -galacturonate to meso -galactarate (mucate). Galactarate has applications in food, cosmetics, pharmaceuticals as a platform chemical. In fungi catabolized through reductive pathway with reductase first enzyme. Deleting corresponding gene Hypocrea jecorina Aspergillus niger resulted unable...

Resistance to weak organic acids is important relative both acid preservatives and the development of inhibitor tolerant yeast as industrial production organisms. The ABC transporter Pdr12 for resistance sorbic propionic acid, but its role in tolerance other with relevance not well established. In this study, strains altered expression PDR12 and/or CMK1, a protein kinase associated post-transcriptional negative regulation Pdr12, were exposed seven acids: acetic, formic, glycolic, lactic,...

ABSTRACT d -Galacturonic acid, the main monomer of pectin, is an attractive substrate for bioconversions, since pectin-rich biomass abundantly available and pectin easily hydrolyzed. l -Galactonic acid intermediate in eukaryotic pathway -galacturonic catabolism, but extracellular accumulation -galactonic has not been reported. By deleting gene encoding dehydratase ( lgd1 or gaaB ) two filamentous fungi, strains were obtained that converted to acid. Both Trichoderma reesei Δ Aspergillus niger...

Turnover of damaged molecules is considered to play a key role in housekeeping cells exposed oxidative stress, and during the progress ageing. In this work, global changes transcriptome were analysed recovery yeast after H 2 O stress. Regarding induced genes, those associated with protein fate most significantly over-represented. addition genes encoding subunits 20S proteasome, related vacuolar proteolysis ( PEP4 LAP4 ), sorting into vacuole, fusion found be induced. The upregulation gene...

The first enzyme in the pathway for l-arabinose catabolism eukaryotic microorganisms is a reductase, reducing to l-arabitol. enzymes catalyzing this reduction are general nonspecific and would also reduce d-xylose xylitol, step catabolism. It not clear whether use different depending on carbon source. Here we show that Aspergillus niger makes of two enzymes. We identified, cloned, characterized an larA, from xyrA. larA up-regulated l-arabinose, while xyrA d-xylose. There however initial...

ABSTRACT The ability of cells to maintain pH homeostasis in response environmental changes has elicited interest basic and applied research prompted the development methods for intracellular measurements. Many traditional provide information at population level thus average values studied cell physiological phenomena, excluding fact that cultures are very heterogeneous. Single-cell analysis, on other hand, offers more detailed insight into variability, thereby facilitating a considerably...

This work describes the development and characterization of a modular synthetic expression system that provides broad range adjustable predictable levels in S. cerevisiae. The works as fixed-gain transcription amplifier, where input signal is transferred via factor (sTF) onto promoter, containing defined core generating output signal. activation based on bacterial LexA-DNA-binding domain, set modified, LexA-binding sites selection domains. We show both experimentally computationally tuning...

This study focused on the performance of dextran producer Leuconostoc citreum as starter culture during 30 days wheat flour type I sourdough propagation (back-slopping). As confirmed by RAPD-PCR analysis, strain dominated throughout procedure, consisting daily fermentations at 20 °C. The sourdoughs were characterized consistent lactic acid bacteria cell density and acidification parameters, reaching pH values 4.0 mild titratable acidity. Carbohydrates consumption remained leading to...

Anthraquinones in the emodin family are produced by bacteria, fungi, and plants. They display various biological activities exploited, e.g., for crop protection, may also be utilized as sustainable, bio-based colorants textile, paints, electronics, cosmetic industries. Anthraquinone pigments from Cortinarius mushrooms have been used artisan dyeing because they stable, colorfast, compatible with methods. However, their chemical synthesis is complex uneconomical, harvesting wild forests...

In filamentous fungi d ‐galactose can be catabolised through the oxido‐reductive and/or Leloir pathway. pathway is converted to ‐fructose in a series of steps where last step oxidation ‐sorbitol by an NAD‐dependent dehydrogenase. We identified sorbitol dehydrogenase gene, sdhA (JGI53356), Aspergillus niger encoding medium chain which involved and catabolism. The gene upregulated presence ‐galactose, galactitol ‐sorbitol. An deletion strain showed reduced growth on was completely abolished....

Abstract Background Anthranilate is a platform chemical used by the industry in synthesis of broad range high-value products, such as dyes, perfumes and pharmaceutical compounds. Currently anthranilate produced via from non-renewable resources. Biological would allow use renewable carbon sources avoid accumulation toxic by-products. Microorganisms produce an intermediate tryptophan biosynthetic pathway. Several prokaryotic microorganisms have been engineered to overproduce but attempts...

l ‐Xylulose reductase is part of the eukaryotic pathway for ‐arabinose catabolism. A previously identified ‐xylulose in Hypocrea jecorina turned out to be not ‘true’ one since it was upregulated during growth on and deletion strain showed no reduced activity but instead lost d ‐mannitol dehydrogenase [17]. In this communication we Aspergillus niger . The gene, lxrA (JGI177736), results a lacking NADPH‐specific having ‐arabinose. purified enzyme had K m 25 mM ν max 650 U/mg.

Citrus processing waste is a leftover from the citrus industry and available in large amounts. Typically, this dried to produce animal feed, but sometimes it just dumped. Its main component peel, which consists mostly of pectin, with D-galacturonic acid as monomer. Aspergillus niger filamentous fungus that efficiently produces pectinases for hydrolysis pectin uses resulting most other components peel growth. We used engineered A. strains were not able catabolise acid, instead converted...

The D-galacturonic acid derived from plant pectin can be converted into a variety of other chemicals which have potential use as chelators, clarifiers, preservatives and plastic precursors. Among these is the deoxy-keto L-galactonic acid, keto-deoxy-L-galactonic or 3-deoxy-L-threo-hex-2-ulosonic acid. keto-deoxy sugars been found to useful precursors for producing further derivatives. Keto-deoxy-L-galactonate natural intermediate in fungal D-galacturonate metabolic pathway, thus...

L-Xylulose reductases belong to the superfamily of short chain dehydrogenases and (SDRs) catalyze NAD(P)H-dependent reduction L-xylulose xylitol in L-arabinose glucuronic acid catabolism. Here we report identification a novel reductase LXR3 fungus Trichoderma reesei by bioinformatic approach combination with functional analysis. LXR3, 31 kDa protein, catalyzes via NADPH is also able convert D-xylulose, D-ribulose, L-sorbose, D-fructose their corresponding polyols. Transcription lxr3...