Production Of Primary Metabolites Research Articles

Bioinformatics deciphers the thebaine biosynthesis pathway in opium poppy: Hub genes, network analysis, and miRNA regulation

Thebaine, a vital precursor in the codeine and morphine pathway, shows promise in addiction treatment. We conducted a comprehensive study on the thebaine biosynthesis pathway in opium poppy, utilizing bioinformatics tools. The dataset comprising the thirteen genes associated with the thebaine biosynthesis pathway was compiled from an extensive review of published literature and validated using the NCBI BLAST tool. Utilizing STRING and Cytoscape, we analyzed gene interactions and visualized the molecular interaction network, respectively. To identify hub proteins, CytoHubba was administered. The Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) at STRING were used for the enrichment analysis of the hub genes. CytoCluster was used to analyze the network in clusters. Promoter regions of hub genes and potential miRNAs were explored using MEME and the psRNATarget database. Hub genes crucial to thebaine biosynthesis were identified, contributing to essential cellular functions like growth, development, stress response, and signal transduction. Metabolic processes emerged as pivotal for thebaine production, indicating a broader role for the thebaine pathway gene network beyond primary metabolite production. Cell component subnetwork genes demonstrated associations with anatomical units, indicating involvement in plant defense responses. Dominant molecular functions drove plant defense responses. KEGG pathway analysis highlighted the significance of metabolic pathways and biosynthesis of secondary metabolites. Cluster analysis emphasized the relevance of the biosynthesis of amino acids, confirming the link between primary and secondary metabolites. Promoter analysis suggested the potential involvement of signal transduction in thebaine production. Hub genes were targeted by 40 miRNAs, suggesting potential novel biomarkers or target genes within the thebaine biosynthesis pathway. Based on the role of miRNAs identified in connection with the hub genes of the thebaine production process, the secondary metabolite pathway of thebaine appears to be associated with several key plant pathways, e.g. growth, development and stress response. However, these findings, based on bioinformatics analysis, warrant further experimental validation and promise to advance our understanding of the biosynthesis of thebaine and its interactions with other genes and metabolic pathways that influence the production of metabolites.

Insights into intraspecific diversity of central carbon metabolites in Saccharomyces cerevisiae during wine fermentation

Saccharomyces cerevisiae is a major actor in winemaking that converts sugars from the grape must into ethanol and CO2 with outstanding efficiency. Primary metabolites produced during fermentation have a great importance in wine. While ethanol content contributes to the overall profile, other metabolites like glycerol, succinate, acetate or lactate also have significant impacts, even when present in lower concentrations. S. cerevisiae is known for its great genetic diversity that is related to its natural or technological environment. However, the variation range of metabolic diversity which can be exploited to enhance wine quality depends on the pathway considered. Our experiment assessed the diversity of primary metabolites production in a set of 51 S. cerevisiae strains from various genetic backgrounds. Results pointed out great yield differences depending on the metabolite considered, with ethanol having the lowest variation. A negative correlation between ethanol and glycerol was observed, confirming glycerol synthesis as a suitable lever to reduce ethanol yield. Genetic groups were linked to specific yields, such as the wine group and high α-ketoglutarate and low acetate yields. This research highlights the potential of using natural yeast diversity in winemaking. It also provides a detailed data set on production of well known (ethanol, glycerol, acetate) or little-known (lactate) primary metabolites.

Effect of lactic acid bacteria by different concentrations of copper based on non-target metabolomic analysis.

Copper (Cu) is an essential element for mammals, but excess intake can have detrimental health consequences. However, Cu is no longer present in the "Limit of Contaminants in Foods" promulgated in 2022. The potential impact of different Cu (II) concentrations on human health remains unclear. In this study, a strain of lactic acid bacteria (LAB), namely, Lactiplantibacillus plantarum CICC 23121 (L23121), was selected as a prebiotic indicator strain to indirectly assess the effects of food-limited Cu (II) concentrations (issued by Tolerance limit of copper in foods in 1994) on the functions of intestinal microbes. We used non-target metabolomics, automatic growth curve detector, scanning electron microscopy (SEM), and Fourier transform infrared spectroscopy (FTIR) to investigate the effects of Cu (II) on L23121. The study revealed shows that the 50% minimum inhibitory concentration (MIC50) of Cu (II) for most lactic acid bacteria was 4 mg/L. At low Cu (II) concentrations (≤ 4 mg/L), the pentose phosphate pathway and pyrimidine metabolism of the lactic acid bacteria were affected, resulting in a decrease in the content of beneficial secondary metabolites and a significant decrease in the cell activity. As Cu (II) concentrations increase (≥ 6 mg/L), the key amino acid and lipid metabolisms were affected, leading to the inhibition of growth and primary metabolite production of the bacteria. Under high concentration of Cu (II) (6 mg/L), the surface adhesion of the bacteria was distorted and covered with significantly large particles, and the functional groups of the cells were significantly shifted. As a probiotic, the abundance of lactic acid bacteria in the intestine is significantly reduced, which will inevitably seriously damage intestinal homeostasis. Thus, to protect human intestinal microbes' health, it is recommended to limit the concentration of Cu in food to less than 4 mg/L.

Chitosan Induces Sorghum Tolerance to Water Deficits by Positively Regulating Photosynthesis and the Production of Primary Metabolites, Osmoregulators, and Antioxidants

Current climate change scenarios require strategies that mitigate the effects of water deficits. Given that chitosan stimulates induced resistance responses, we tested the hypothesis that foliar application of this polysaccharide can have positive effects on the primary metabolite, antioxidant, and osmoregulatory metabolisms of sorghum, mitigating the effects of water deficit on grain yield. Sorghum plants were cultivated under either irrigated or water-deficit conditions and were either treated or untreated with chitosan, and we assessed the following responses: water potential, gas exchange, nutritional status, biosynthesis of primary and osmoregulatory metabolites, the activity of the antioxidant system constituents, and hydrogen peroxide and malondialdehyde contents. An active methodology was used to impose water stress on plants grown in pots with 20 kg of soil, 06 replications per treatment. Sorghum plants subjected to water deficit that were treated with chitosan exhibited higher stomatal conductance and transpiration than the untreated plants. Water restriction reduced the nutrient contents of leaves, but in the irrigated plants, chitosan treatment increased the tissue contents of certain nutrients and leaf concentrations of total proteins and total amino acids. In the plants under water-deficit conditions that were treated with chitosan, an elevated photosynthetic rate contributed to an increase in carbohydrate accumulation and a higher production of panicle and grain dry biomass compared with the untreated plants. The activity of catalase and ascorbate peroxidase also increased in response to chitosan. Overall, chitosan was effective in enhancing the water deficit tolerance of sorghum plants cultivated under water-deficit conditions by 57%. With this work, we generate perspectives for studies focusing on the development of water-deficit-resistant sorghum lineages, by means of chitosan elicitation.

Production of Primary Metabolites by Rhizopus stolonifer, Causal Agent of Almond Hull Rot Disease.

Species in the fungal genus Rhizopus are able to convert simple sugars into primary metabolites such as fumaric acid, lactic acid, citric acid, and, to a lesser extent, malic acid in the presence of specific carbon and nitrogen sources. This ability has been linked to plant pathogenicity. Rhizopus stolonifer causes hull rot disease in almonds, symptoms of which have been previously associated with the fungus's production of fumaric acid. Six isolates of R. stolonifer taken from infected almond hulls were grown in artificial media amended with one of four carbon sources (glucose, fructose, sucrose, and xylose) and two nitrogen sources (asparagine and ammonium sulphate) chosen based on almond hull composition and used in industry. Proton nuclear magnetic resonance (1H NMR)-based metabolomics identified that R. stolonifer could metabolise glucose, fructose, sucrose, and to a lesser extent xylose, and both nitrogen sources, to produce three metabolites, i.e., fumaric acid, lactic acid, and ethanol, under in vitro conditions. Sugar metabolisation and acid production were significantly influenced by sugar source and isolates, with five isolates depleting glucose most rapidly, followed by fructose, sucrose, and then xylose. The maximum amounts of metabolites were produced when glucose was the carbon source, with fumaric acid produced in higher amounts than lactic acid. Isolate 19A-0069, however, preferred sucrose as the carbon source, and Isolate 19A-0030 produced higher amounts of lactic acid than fumaric acid. This is the first report, to our knowledge, of R. stolonifer producing lactic acid in preference to fumaric acid. Additionally, R. stolonifer isolate 19-0030 was inoculated into Nonpareil almond fruit on trees grown under high- and low-nitrogen and water treatments, and hull compositions of infected and uninfected fruit were analysed using 1H NMR-based metabolomics. Glucose and asparagine content of uninfected hulls was influenced by the nitrogen and water treatments provided to the trees, being higher in the high-nitrogen and water treatments. In infected hulls, glucose and fructose were significantly reduced but not sucrose or xylose. Large amounts of both fumaric and lactic acid were produced, particularly under high-nitrogen treatments. Moreover, almond shoots placed in dilute solutions of fumaric acid or lactic acid developed leaf symptoms very similar to the 'strike' symptoms seen in hull rot disease in the field, suggesting both acids are involved in causing disease.

Study of Epilobium angustifolium L. amino acids content by HPLC method

The use of plant raw materials is one of the areas of modern pharmaceutical science in the production of herbal drugs. The genus Epilobium counts more than 200 species, many species of which are used in traditional medicine. Among the Epilobium species, Epilobium angustifolium is one of the well-known medicinal plants which have been used worldwide in habitual medicine. There is insufficient information in the literature on the biologically active substances of Epilobium angustifolium L. The presence of three major polyphenol groups: phenolic acids, flavonoids, and ellagitannins were identified in E. angustifolium extracts. Traditionally, the infusion of leaves of this plant could be useful for headaches, cold and gastrointestinal disorder. The Epilobium angustifolium L. as an insufficiently studied plant is a promising object of study, including amino acids composition. To assess the relationship between the production of primary metabolites and their possible therapeutic properties, we analyzed the amino acid profile of the plant Epilobium angustifolium used in traditional medicine. The study of compounds generated by plants as a result of defense mechanisms permits an understanding of the molecular mechanism involved in their medicinal properties.
 The aim. Thus, the aim of the study was to conduct an HPLC analysis of the amino acids of E. angustifolium to establish the prospects for the use of the raw materials in medical and pharmaceutical practice. The results of the current study will be used in further breeding programs aimed to obtain an industrial form of E. angustifolium suitable for pharmaceutical and food applications.
 Materials and methods. The determination of amino acids composition of Epilobium angustifolium was conducted using Agilent 1200 (Agilent Technologies, USA).
 Results. The HPLC method identified sixteen free amino acids and seventeen bound amino acids in the Epilobium angustifolium herb. The studies have shown that Epilobium angustifolium L. herb is mainly composed of free amino acids such as L-phenylalanine (1.65 µg/mg), L-glutamic acid (1.51 µg/mg), L-arginine (1.24 µg/mg), L-alanine (0.98 µg/mg) and L-aspartic acid (0.57 µg/mg), which were presents in the greatest amount. The dominant bound amino acids in the studied raw material were L-glutamic acid, L-aspartic acid, L-leucine, and L-alanine, the content of which was 32.37 µg/mg, 10.59 µg/mg, 8.70 µg/mg, and 6.22 µg/mg respectively.
 Conclusions. Using the HPLC method determined the amino acids in the herb of Epilobium angustifolium L. The concentrations of L-aspartic acid, L-glutamic acid, L-arginine, L-alanine and L-phenylalanine are predominate among free and bound amino acids in the Epilobium angustifolium L. herb. The result shows that Epilobium angustifolium L. is the source of amino acids, so the use of this plant raw material for new remedies is possible in the future

Relationship between lipopeptide biosurfactant and primary metabolite production by Bacillus strains in solid-state and submerged fermentation

The relationship between lipopeptide and primary metabolite production by Bacillus spp. in solid-state fermentation (SSF) and submerged fermentation (SmF) was evaluated. Four wild-type strains and one mutant strain (unable to develop biofilm) were assessed in SSF and SmF, using a defined medium and polyurethane foam as inert support for SSF. Strain ATCC 21,332 in SSF presented the highest lipopeptide production. The wild-type strains revealed higher lipopeptide and biomass production and lower synthesis of primary metabolites in SSF than in SmF. However, the mutant strain showed a slightly higher production of primary metabolites in SSF than in SmF. Carbon balance analysis showed that the carbon flux was mainly directed to lipopeptides in SSF, whereas in SmF, it was directed to the production of primary metabolites and the carbon flux to lipopeptides is inversely related to primary metabolites in both types of cultures.

The effect of Aceh’s natural stone presence on the growth, produced biomass and primary metabolites production of Chlorella vulgaris LIPI12-AL042

This study sought to evaluate the influence of natural stone presence that was collected from Aceh Province, on the growth and the production of biomass, protein, total sugar and lipid of the microalgae Chlorella vulgaris LIPI12-AL042. Aceh’s natural stone used in this study were nephrite jade (NJ) and black jade (BJ) which were known to possess different minerals composition. The cellular growth of microalgae in terms of biomass yield and its productivity increased significantly (13,38%) under the presence of BJ stone compared to the control. Different results of reduced biomass yield and its productivity were demonstrated by the microalgae culture under the influence of NJ stone’s presence compared to the control. In terms of total sugar, both stones exhibited superior results over the control. The protein content and its yield of the Chlorella vulgaris LIPI12-AL042 cultivated under the influence of the stones demonstrated less quantity over the control.

Indoor mass cultivation of marine diatoms for biodiesel production using induction plasma synthesized nanosilica

In this work, two benthic marine diatoms Chaetoceros sp. and Thalassiosira sp. were grown in modified f/2 medium in which normal silica was replaced with inductively coupled plasma (ICP) nanosilica for indoor mass cultivation and its impact on growth, lipid content, lipid quality and metabolite production were monitored. Results indicate thatunder mass cultivation using ICP nano silica medium, Thalassiosirasp. reached 3.6 and Chaetoceros sp. reached 3.2-fold higher cell density compared to normal Si medium. The primary metabolite production and total lipid content was higher in Chaetoceros sp. (44.33 ± 2.51% DCW) compared to Thalassiosira sp. (29.66 ± 1.52% DCW). In mass cultivation, ICP synthesized nanosilica powder was effective in enhancing the cell density, production of metabolites, pigments, and lipids in the marine diatoms studied. This is the first report on the use of ICP nanosilica in carrying out indoor mass cultivation of marine diatom isolates as potential biodiesel and biomolecule feedstocks.

Optimization of Cellulose Production by Curvularia pallescens Isolated from Textile Effluent

Introduction: Celluloses are important industrial enzymes and find application in several industrial processes. Effects of pH, temperature, incubation time, source of carbon and nitrogen were tested in submerged fermentation process in the production of cellulose by Curvularia pallescens isolated from textile effluent.
 Aims: The present study was attempted in a fungus; Curvularia pallescens isolated from textile effluent for maximizing its production under optimal conditions in submerged fermentation by using inexpensive substrate wheat bran.
 Study Design: The production medium was prepared in distilled water, supplemented with 4.5% wheat bran, 0.05% KCl, 0.2% KH2PO4, (carbon source), yeast extract (nitrogen source), maintained with pH of 5.5 and incubated at 28ºC for 120 h was found optimal for the production of cellulose.
 Results: The test fungus achieved maximum FPA activity followed by cellobiohydrolase, endoglucanase and β-glucosidase activity at 46.76, 42.06, 26.94 and 3.56 U/ml respectively at pH 5.5 (Fig. 4). The temperature of 280C produced maximum cellulase activity. Highest activity recorded was of FPA (38.94 U/ml), followed by cellobiohydrolase (30.29 U/ml), endoglucanase (22.41 U/ml), and β-glucosidase (3.98 U/ml). The effect of process parameters such as the effect of temperature, pH and inoculum size was investigated. Maximum cellulase and xylanase having an enzyme activity of 694.45 and 931.25 IU, respectively, were produced at 30ºC incubation temperature.
 Conclusion: The effect of process parameters such as effect of temperature, pH and inoculum size was also investigated. The production of primary metabolites by microorganisms is highly influenced by their growth, which is determined by the availability of the nutrients in the substrates.

Aktivitas Mikrob dalam Pulp Biji Kakao (Theobroma cacao L.) Selama Fermentasi dengan Penambahan Ragi Tape

<em>Microbial activities in cocoa pulp decomposition is key in cocoa beans fermentation. </em>Ragi tape<em> is widely used </em><em>as a source of microbes (starters). The study aimed to investigate the pattern of changes in the number of microbes, substrate degradation activities and primary metabolites production during fermentation with the addition of ragi tape. The study was conducted at the laboratories within the Faculty of Agriculture Technology and Integrated Laboratory of Experiment and Research, UGM from March to October 2018 . Fermentation experiment used 2 kg of fresh cacao beans stored in a plastic box per batch, its temperature was set daily. The amount of microbes was enumerated using total plate count (TPC), whereas substrate degradation and primary metabolites were analyzed using high performance liquid chromatography (HPLC) and gas chromatography (GC). Temperature, pulp and seed pH, and fermentation index were observed. The results showed that adding </em>ragi tape<em> </em><em>for fermentation generated a maximum amount of yeast, lactic acid bacteria (LAB) and acetic acid bacteria (AAB) of 9.03<span style="text-decoration: underline;">+</span>0.85; 9.05<span style="text-decoration: underline;">+</span>0.17; and 9.15<span style="text-decoration: underline;">+</span>0.89 log cfu/g of cocoa pulp respectively, with substrate degradation in the form of sucrose reduced by 97%, glucose 98.6%, fructose 97%, and citric acid 71% on the third day. Maximum production of primary metabolites of ethanol is 27.84<span style="text-decoration: underline;">+</span>21.85 mg/g of cocoa pulp, lactic acid 4.18<span style="text-decoration: underline;">+</span>3.16 mg/g of cocoa pulp and acetic acid 3.38<span style="text-decoration: underline;">+</span>5.43 mg/g of cocoa pulp. Fermentation with the addition of </em>ragi tape<em> accelerates the process to three days with a fermentation index value of 1.05<span style="text-decoration: underline;">+</span>0.06 and seed pH of 5.97<span style="text-decoration: underline;">+</span>0.20.</em>

An Overview of Sucrose Synthases in Plants.

Sucrose is the end product of photosynthesis and the primary sugar transported in the phloem of most plants. Sucrose synthase (SuSy) is a glycosyl transferase enzyme that plays a key role in sugar metabolism, primarily in sink tissues. SuSy catalyzes the reversible cleavage of sucrose into fructose and either uridine diphosphate glucose (UDP-G) or adenosine diphosphate glucose (ADP-G). The products of sucrose cleavage by SuSy are available for many metabolic pathways, such as energy production, primary-metabolite production, and the synthesis of complex carbohydrates. SuSy proteins are usually homotetramers with an average monomeric molecular weight of about 90 kD (about 800 amino acids long). Plant SuSy isozymes are mainly located in the cytosol or adjacent to plasma membrane, but some SuSy proteins are found in the cell wall, vacuoles, and mitochondria. Plant SUS gene families are usually small, containing between four to seven genes, with distinct exon-intron structures. Plant SUS genes are divided into three separate clades, which are present in both monocots and dicots. A comprehensive phylogenetic analysis indicates that a first SUS duplication event may have occurred before the divergence of the gymnosperms and angiosperms and a second duplication event probably occurred in a common angiosperm ancestor, leading to the existence of all three clades in both monocots and dicots. Plants with reduced SuSy activity have been shown to have reduced growth, reduced starch, cellulose or callose synthesis, reduced tolerance to anaerobic-stress conditions and altered shoot apical meristem function and leaf morphology. Plants overexpressing SUS have shown increased growth, increased xylem area and xylem cell-wall width, and increased cellulose and starch contents, making SUS high-potential candidate genes for the improvement of agricultural traits in crop plants. This review summarizes the current knowledge regarding plant SuSy, including newly discovered possible developmental roles for SuSy in meristem functioning that involve sugar and hormonal signaling.

Production of primary metabolites in Microcystis aeruginosa in regulation of nitrogen limitation

The aim of this work was to study the regulatory effect of nitrogen (N) deficiency on primary metabolites in Microcystis aeruginosa, and promote the utilization of the alga. Low-N and Non-N conditions, especially Non-N, reduced the cell growth and photosynthetic abilities compared to Normal-N, as N deficiency triggered the down-regulation of genes involving in the photosynthetic process. Non-N not changed lipid content, due to no up-regulation of genes that promoted lipid synthesis. Soluble protein content significantly decreased under Non-N, which may result from the declined expression of genes relating to amino acid and histidyl-transfer RNA synthesis. Soluble and insoluble carbohydrate content significantly increased under Non-N, as the expression variation of genes blocked sugar degradation and promoted lipopolysaccharide synthesis. Therefore, M. aeruginosa can be used as the feedstock to produce carbohydrates under N deficiency for bioethanol production, and the remainder lipids after carbohydrate extraction can be used to produce biodiesel.

The impact of culture conditions on growth and metabolomic profiles of freshwater cyanobacteria.

Cultured cyanobacteria produce secondary metabolites with a wide range of biological activities and are an important source of natural products. In the context of secondary metabolite discovery, microbial culture conditions are expected to support optimum growth, induce maximum chemical diversity, and be suitable for the majority of cyanobacterial strains. We investigated the effect of nitrate and phosphate on biomass production and metabolomic profiles of three filamentous freshwater cyanobacterial strains: cf. Oscillatoria sp. UIC 10045, Scytonema sp. UIC 10036, and Nostoc sp. UIC 10110. A standardized inoculation procedure allowed for the assessment of cell mass production. Dried cyanobacterial cell mass was extracted and analyzed by liquid chromatography coupled with high resolution mass spectrometry (UPLC-HRMS), followed by comparative metabolomics analysis using XCMS Online. Results showed that low nitrate media significantly reduced cell mass production for all three strains. Low nitrate also induced production of primary metabolites (heterocyst glycolipids) in strains UIC 10036 and UIC 10110. Changes in phosphate levels affected each strain differently. Strain UIC 10110 showed a significant increase in production of merocyclophane C when cultivated in low phosphate, while strain UIC 10036 displayed higher production of tolytoxin under high phosphate. Additionally, these experiments led to the identification of a potentially new peptide produced by strain UIC 10036.

Herbivory-induced maternal effects on growth and defense traits in the clonal species Alternanthera philoxeroides

Plants have evolved a variety of defense traits against foliar herbivory, including the production of primary and secondary metabolites, the allocation of chemical compounds, and morphological plasticity. Using two vegetative generations of the invasive clonal species Alternanthera philoxeroides, we investigated the effects of maternal and offspring herbivory by Planococcus minor on the integrative defense strategy of plants. Herbivory severely inhibited leaf, stolon and root growth, as well as the production of primary metabolites (soluble sugars, starch, and total non-structural carbohydrates in stolons), and decreased average leaf area and specific leaf area of the second-generation A. philoxeroides. The changes in growth measures of the first-generation A. philoxeroides with herbivory were consistent with that of the second generation. By contrast, herbivory basically did not affect the concentration of non-structural carbohydrate compounds in the roots, and even increased the concentrations of N and total phenols in taproots. Furthermore, herbivory-induced maternal effects also reduced the growth of the second-generation plants. The results suggest that A. philoxeroides is capable of adapting to herbivory by P. minor, mainly via the allocation of available resources in belowground organs, and that the herbivory effect can persist across vegetative generations. These features may potentially improve the regeneration and tolerance of A. philoxeroides after a short-term herbivory.

Biomanufacturing: history and perspective.

Biomanufacturing is a type of manufacturing that utilizes biological systems (e.g., living microorganisms, resting cells, animal cells, plant cells, tissues, enzymes, or in vitro synthetic (enzymatic) systems) to produce commercially important biomoleculesfor use inthe agricultural, food, material, energy, and pharmaceutical industries. History of biomanufacturing could be classified into the three revolutions in terms of respective product types (mainly), production platforms, and research technologies. Biomanufacturing 1.0 focuses on the production of primary metabolites (e.g., butanol, acetone, ethanol, citric acid) by using mono-culture fermentation; biomanufacturing 2.0 focuses on the production of secondary metabolites (e.g., penicillin, streptomycin) by using a dedicated mutant and aerobic submerged liquid fermentation; and biomanufacturing 3.0 focuses on the production of large-size biomolecules-proteins and enzymes (e.g., erythropoietin, insulin, growth hormone, amylase, DNA polymerase) by using recombinant DNA technology and advanced cell culture. Biomanufacturing 4.0 could focus on new products, for example, human tissues or cells made by regenerative medicine, artificial starch made by in vitro synthetic biosystems, isobutanol fermented by metabolic engineering, and synthetic biology-driven microorganisms, as well as exiting products produced by far better approaches. Biomanufacturing 4.0 would help address some of the most important challenges of humankind, such as food security, energy security and sustainability, water crisis, climate change, health issues, and conflict related to the energy, food, and water nexus.

Metabolite transport and process enhancement in cell factories

Cell factories are the basis of biorefinery and necessary for the development and application of bio-based resources. Metabolite transport is essential for cell metabolism as well as the synthesis of diverse metabolites. There are a variety of methods to transport metabolites, with different transport efficiencies. A low transport efficiency greatly affects the production of cell factories. The two major transport methods can be defined as transport with or without membranes, which can be modified and optimized to increase the production of primary metabolites, secondary metabolites, and biomacromolecules in cell factories. Synthetic biology and metabolic engineering methods can be used to modify or redesign and construct transport pathways, which can effectively enhance metabolite transport, reduce the loss of intermediate metabolites, and increase the yield of products. Studies on the usage of metabolite transport to enhance the production of cell factories were reviewed, and references and methods for future systematic studies of metabolite transport were identified.

Butyrate influences intracellular levels of adenine and adenine derivatives in the fungus Penicillium restrictum

Butyrate, a small fatty acid, has an important role in the colon of ruminants and mammalians including the inhibition of inflammation and the regulation of cell proliferation. There is also growing evidence that butyrate is influencing the histone structure in mammalian cells by inhibition of histone deacetylation. Butyrate shows furthermore an antimicrobial activity against fungi, yeast and bacteria, which is linked to its toxicity at a high concentration. In fungi there are indications that butyrate induces the production of secondary metabolites potentially via inhibition of histone deacetylases. However, information about the influence of butyrate on growth, primary metabolite production and metabolism, besides lipid catabolism, in fungi is scarce. We have identified the filamentous fungus Penicillium (P.) restrictum as a susceptible target for butyrate treatment in an antimicrobial activity screen. The antimicrobial activity was detected only in the mycelium of the butyrate treated culture. We investigated the effect of butyrate ranging from low (0.001mM) to high (30mM), potentially toxic, concentrations on biomass and antimicrobial activity. Butyrate at high concentrations (3 and 30mM) significantly reduced the fungal biomass. In contrast P. restrictum treated with 0.03mM of butyrate showed the highest antimicrobial activity. We isolated three antimicrobial active compounds, active against Staphylococcus aureus, from P. restrictum cellular extracts treated with butyrate: adenine, its derivate hypoxanthine and the nucleoside derivate adenosine. Production of all three compounds was increased at low butyrate concentrations. Furthermore we found that butyrate influences the intracellular level of the adenine nucleoside derivate cAMP, an important signalling molecule in fungi and various organisms.In conclusion butyrate treatment increases the intracellular levels of adenine and its respective derivatives.

Enhancement of Surfactin and Fengycin Production by Bacillus mojavensis A21: Application for Diesel Biodegradation.

This work concerns the study of the enhancement of surfactin and fengycin production by B. mojavensis A21 and application of the produced product in diesel biodegradation. The influences of the culture medium and cells immobilization were studied. The highest lipopeptides production was achieved after 72 hours of incubation in a culture medium containing 30 g/L glucose as carbon source and a combination of yeast extract (1 g/L) and glutamic acid (5 g/L) as nitrogen sources with initial pH 7.0 at 30°C and 90% volumetric aeration. The study of primary metabolites production showed mainly the production of acetoin, with a maximum production after 24 h of strain growth. The use of immobilized cells seemed to be a promising method for improving lipopeptides productivity. In fact, the synthesis of both lipopeptides, mainly fengycin, was greatly enhanced by the immobilization of A21 cells. An increase of diesel degradation capacity of approximately 20, 27, and 40% in the presence of 0.5, 1, and 2 g/L of produced lipopeptides, respectively, was observed. Considering these properties, B. mojavensis A21 strain producing a lipopeptide mixture, containing both surfactin and fengycin, may be considered as a potential candidate for future use in bioremediation and crop protection.

Opportunities and limitations for the production of safe fermented meats without nitrate and nitrite using an antibacterial Staphylococcus sciuri starter culture

The ability of the meat isolate Staphylococcus sciuri I20-1 to inhibit food-borne pathogens in view of its application as a functional starter culture for the production of clean-label fermented meats was investigated. The strain produced a heat-stable antibacterial compound of a proteinaceous nature that was released following primary metabolite production kinetics. Its specific inhibitory spectrum included different strains of Staphylococcus aureus and both vegetative cells and spores of Clostridium botulinum. These two species pose potential biosafety hazards, in particular when producing fermented meats with a mild pH and without added nitrate and nitrite salts. Up till now, starter-culture based strategies to restrain S. aureus and C. botulinum have barely been explored in fermented meats, in contrast to the frequently investigated use of antilisterial cultures of lactic acid bacteria. When added to a fermented sausage model, S. sciuri I20-1 acted as a persistent culture throughout the fermentation process, during which it was able to produce its antibacterial compound in situ. In co-culture experiments, an immediate two-log inactivation of a S. aureus strain was found, even before production of the antibacterial compound could be detected. This was probably due to an instant pH-driven desorption of the antibacterial molecules from the producer cells when added to the meat batter. However, a sub-population of S. aureus that was resistant to the antibacterial compound was able to grow out to the same final counts as in the control experiment performed with a non-inhibitory strain of S. sciuri. This finding is indicative of the potential pitfalls when extrapolating promising in vitro results to the true complexity of fermented food matrices.