3-hydroxypropionic Acid Research Articles

Production of 3-hydroxypropionic acid from acetate using metabolically-engineered and glucose-grown Escherichia coli

Acetate can be used as carbon feedstock for the production of 3-hydroxypropionic acid (3-HP), but the production level was low due to inefficient cell growth on acetate. To better utilize acetate, a two-stage strategy, whereby glucose is used for cell growth and acetate for 3-HP formation, was attempted. Dissected malonyl-CoA reductase of Chloroflexus aurantiacus, alone or along with acetyl-CoA carboxylase and/or transhydrogenase, was overexpressed, and by-products formation pathway, glyoxylate shunt (GS) and gluconeogenesis were modified. When GS or gluconeogenesis was disrupted, cell growth on glucose was not hampered, while on acetate it was completely abolished. Consequently, 3-HP production, at production stage, were low, though 3-HP yield on acetate was increased, especially in the case of aceA deletion. In two-stage bioreactor, strain with upregulated GS produced 7.3 g/L 3-HP with yield of 0.26 mol/mol acetate. This study suggests that two-stage cultivation is a good strategy for 3-HP production from acetate.

In Vitro Antimicrobial and Antioxidant Activities of Lactobacillus coryniformis BCH-4 Bioactive Compounds and Determination of their Bioprotective Effects on Nutritional Components of Maize (Zea mays L.).

Lactic acid bacteria (LAB) can synthesize antimicrobial compounds (AMCs) with nutritional and bioprotective properties in crops and food products. In the current study, AMCs of Lactobacillus coryniformis BCH-4 were evaluated to control fungal spoilage in maize grains. On maize grains treated with 75%–100% (v/v) concentrated AMCs, no fungal growth was observed even after 72 h of Aspergillus flavus inoculation. Proximate analysis of treatments A1 (raw grains), A2 (A. flavus inoculated grains) and A3 (A. flavus + AMCs inoculated grains) revealed that moisture was significantly (p ≤ 0.05) high in A2 than A3 and A1. Meanwhile, protein, fat, fiber and ash contents were significantly decreased in A2 compared to A1 and A3. Moreover, β-carotene contents were not statistically different between A1 and A3, while in A2 it was significantly decreased. HPLC analysis revealed the presence of 2-oxopropanoic acid, 2-hydroxypropane-1,2,3-tricarboxylic acid, 2-hydroxybutanedioic acid, 2-hydroxypropanoic acid, propanedioic acid and butanedioic acid, which also showed antifungal activity against Aspergillus flavus. FTIR spectroscopy revealed the presence of hydroxyl, carbonyl and ester-groups along with organic and fatty acids, thereby indicating their participation in inhibitory action. Furthermore, the AMCs were found to be a good alternative to chemical preservatives, thereby not only preserving the nutritive qualities but increasing the shelf life as well.

Biochemical and genetic approaches to the prenatal diagnosis of propionic acidemia in 78 pregnancies

BackgroundPropionic acidemia (PA) is a serious metabolic disorder, and different approaches have been applied to its prenatal diagnosis. To evaluate the reliability and validity of a biochemical strategy in the prenatal diagnosis of PA, we conducted a retrospective study of our 11-year experiences at a single center.MethodsWe accumulated data from 78 pregnancies from 58 families referred to our center and provided prenatal diagnosis by directed genetic analysis and/or metabolite measurement using tandem mass spectrometry (MS/MS) and gas chromatography/mass spectrometry (GC/MS) of amniotic fluid (AF) samples.ResultsSixty-five unaffected fetuses (83.33%) and 13 affected fetuses (16.67%) were confirmed in our study. The characteristic metabolites including propionylcarnitine (C3) level, C3/acetylcarnitine (C2) ratio and 2-methylcitric acid (2MCA) level in unaffected and affected groups showed significant differences (P < 0.0001), while the level of 3-hydroxypropionic acid (3HPA) showed no significant difference between the two groups (P > 0.05).Of the 78 pregnancies, 24 fetuses were found to have either one causative pathogenic variant or were without genetic information in the proband. Three of these fetuses had elevated AF levels of C3, C3/C2 ratio, and 2MCA and, thus, were determined to be affected, while the remaining fetuses were determined to be unaffected based on a normal AF metabolite profile. Our genetic and biochemical results were highly consistent with postnatal follow-up results on all unaffected fetuses.ConclusionsWe conclude that a biochemical approach can serve as a fast and convenient prenatal diagnostic method for pregnancies at an increased risk for PA, which could be used in conjunction with genetic testing for precise prenatal diagnosis of this disorder. In our analysis, the characteristic metabolites C3 level, C3/C2 ratio, and 2MCA level in AF supernatant were dependable biochemical markers for diagnosis, of which the C3/C2 ratio appears to be the most reliable biochemical marker for the prenatal diagnosis of PA.

Intensifying niacin-based biosynthesis of NAD+ to enhance 3-hydroxypropionic acid production in Klebsiella pneumoniae.

Glycerol-based biosynthesis of 3-hydroxypropionic acid (3-HP) in Klebsiella pneumoniae involves two reactions: glycerol conversion to 3-hydroxypropionaldehyde (3-HPA) by glycerol dehydratase, and 3-HPA conversion to 3-HP by aldehyde dehydrogenase (ALDH). The ALDH catalysis consumes a lot of cofactor nicotinamide adenine dinucleotide (NAD+), which constrains 3-HP production. Here we report that intensifying niacin-based biosynthesis of NAD+ can substantially enhance 3-HP production. We constructed tac promoter-driven NAD+ synthesis pathway in K. pneumoniae. The strain only overexpressing nicotinate phosphoribosyltransferase (PncB) showed 14.24% increase in the production of NAD+ relative to the stain harboring an empty vector. When PncB was coexpressed with PuuC (one of native ALDHs), the recombinant strain exhibited increased ALDH activity but slightly reduced 3-HP production due to plasmid burden. When 30mg niacin l-1 (a substrate for biosynthesis of NAD+) was added into shake flask, the strain produced 0.55g 3-HP l-1, which was 2.75 times that of the control. In a 5-L bioreactor, replenishment of niacin led to 36.43% increase of 3-HP production. These results indicated that intensifying niacin-based biosynthesis of NAD+ boosts 3-HP production.

Current advance in biological production of short-chain organic acid.

As natural metabolites, organic acids have been widely applied in food, pharmaceutical, and bio-based materials industries. Particularly, the short-chain organic acids, including C2, C3, C4, C5, and C6 organic acids, are necessary intermediate metabolites in cells and are also alternatives to some commercial chemical products. As the necessary metabolites in cells, most major short-chain organic acids can be produced through microbial fermentation. Specifically, with the development of synthetic biology, metabolic engineering could endow cells with the ability to produce more short-chain organic acid products including propionic acid, pyruvate, lactic acid, 3-hydroxypropionic acid, malic acid, succinic acid, fumaric acid, butyric acid, itaconic acid, α-ketoglutaric acid, glutaric acid, citric acid, gluconic acid, muconic acid, adipic acid, xylonic acid, and so on. The recent advances in the biological production of short-chain organic acids, as well as the challenges and perspectives, are summarized in this review to promote the generation of microbial cell factories for the production of short-chain organic acids.Key points• Outlines the production strategy of short-chain organic acids• Provide guidance for efficient synthesis of short-chain organic acids• Impacts the necessary factor of acid resistance on the successful production of host cells.

Promiscuous phosphoketolase and metabolic rewiring enables novel non-oxidative glycolysis in yeast for high-yield production of acetyl-CoA derived products

Carbon-conserving pathways have the potential of increasing product yields in biotechnological processes. The aim of this project was to investigate the functionality of a novel carbon-conserving pathway that produces 3 mol of acetyl-CoA from fructose-6-phosphate without carbon loss in the yeast Saccharomyces cerevisiae. This cyclic pathway relies on a generalist phosphoketolase (Xfspk), which can convert xylulose-5-phosphate, fructose-6-phosphate and sedoheptulose-7-phosphate (S7P) to acetyl phosphate. This cycle is proposed to overcome bottlenecks from the previously reported non-oxidative glycolysis (NOG) cycle. Here, in silico simulations showed accumulation of S7P in the NOG cycle, which was resolved by blocking the non-oxidative pentose phosphate pathway and introducing Xfspk and part of the riboneogenesis pathway. To implement this, a transketolase and transaldolase deficient S. cerevisiae was generated and a cyclic pathway, the Glycolysis AlTernative High Carbon Yield Cycle (GATHCYC), was enabled through xfspk expression and sedoheptulose bisphosphatase (SHB17) overexpression. Flux through the GATHCYC was demonstrated in vitro with a phosphoketolase assay on crude cell free extracts, and in vivo by constructing a strain that was dependent on a functional pathway to survive. Finally, we showed that introducing the GATHCYC as a carbon-conserving route for 3-hydroxypropionic acid (3-HP) production resulted in a 109% increase in 3-HP titers when the glucose was exhausted compared to the phosphoketolase route only.

Biotransformation of two citrus flavanones by lactic acid bacteria in chemical defined medium.

Microbial processes are being developed to transform flavonoid glycosides to varieties of metabolites with higher bioavailability. The aim of this study was to determine the metabolic activity and survival of five lactic acid bacteria (LAB) stains (L. rhamnosus LRa05, L. casei LC89, L. plantarum N13, L. acidophilus LA85, and L. brevis LB01) in two different citrus flavanone standards (hesperetin-7-O-rutinoside and naringenin-7-O-rutinoside). The enzymatic activity, metabolites, antioxidant activities, and α-glucosidase inhibition property in the two standards were also investigated before and after incubated with LAB. The medium contained standards permitted survival of the five LAB stains. All strains exhibited β-glucosidase activity. Of the five LAB strains tested, just L. plantarum N13 and L. brevis LB01 have the ability to metabolize hesperetin-7-O-rutinoside, only L. plantarum N13, L. acidophilus LA85, and L. brevis LB01 could metabolize naringenin-7-O-rutinoside, moreover, L. acidophilus LA85l was the strain with the highest biotransformation ratio of naringenin-7-O-rutinoside. L. acidophilus LA85 and L. plantarum N13 can degrade naringenin-7-O-rutinoside into naringenin. L. brevis LB01 can degrade hesperetin-7-O-rutinoside into hesperetin, 3-(4'-hydroxyphenyl)-2-propenoic acid, 3-(3'-hydroxy-4'-methoxyphenyl)hydracrylic acid, and 3-(4'-hydroxyphenyl)propionic acid. Incubation of L. acidophilus LA85 in naringenin-7-O-rutinoside solution supposed no apparent influence in the biological activities that tested. L. acidophilus LA85 may potentially contribute to the bioavailability of citrus flavanones, and to be applied as functional cultures to obtain more bioavailable and bioactive metabolites in food products or in the human gastrointestinal tract.

Serum metabolomics reveals the effect of electroacupuncture on urinary leakage in women with stress urinary incontinence

Stress urinary incontinence (SUI), which is defined as an involuntary loss of urine upon physical exertion coughing, sneezing or laughing, has a significant negative impact on the quality of life of many women. Multi-center, large-scale and randomized clinical trials have illustrated that non-invasive electroacupuncture is an effective treatment for SUI, but its therapeutic mechanism in treating SUI remains unknown. Here, gas chromatography-mass spectrometry based serum metabolomics was performed to reveal metabolic profiles and diagnostic biomarkers from recruitment of 25 patients and 25 healthy women before and after electroacupuncture. We identified 10 differentially abundant metabolites, including butantriol, 3,4-dihydroxybutanoic acid, succinic acid, 1-deoxypentitol, psicose, citric acid, 3-hydroxybutyric acid, hydracrylic acid, 3-hydroxyphenylacetic acid and D-mannitol, from patients between before and after electroacupuncture. The electroacupuncture therapy altered propanoate metabolism, butanoate metabolism and the tricarboxylic acid cycle. A panel of 8 biomarkers (butantriol, 3,4-dihydroxybutanoic acid, succinic acid, 1-deoxypentitol, psicose, citric acid, 3-hydroxybutyric acid and hydracrylic acid) was evaluated to determine the effect of electroacupuncture on SUI and differentiated well between before and after treatment. The area under the receiver operating characteristic curve was 0.962. The sensitivity, specificity and coincidence rate were 92 %, 92 % and 96 %, respectively, at a 95 % confidence interval ranging from 0.9053 to 1. Furthermore, the levels of these diagnostic biomarkers were not significantly altered in healthy subjects after sham electroacupuncture. It was indicated that an 8-biomarker panel might be constructed for the therapeutic evaluation of electroacupuncture treatment for SUI.

Progress in metabolic engineering of biosynthesis of 3-hydroxypropionic acid

As an important platform compound, 3-hydroxypropionic acid (3-HP) can be used as a substrate to synthesize a variety of biological products with commercial potential. The titer of 3-HP by wild-type bacteria is low, which severely limits the large-scale application and production of 3-HP. By modifying the genes related to the metabolic pathway, engineered bacteria using cheap substrates as carbon sources are constructed, the aim of reducing production cost and increasing output is realized. In this paper, the recent progress in the synthesis of 3-HP by metabolic engineering at home and abroad is reviewed. The advantages and disadvantages of glycerol pathway, malonyl-CoA pathway and beta-alanine pathway for synthesis of 3-HP are also summarized and analyzed, and the future development of 3-HP is prospected.

Bioactive hydroxypropionylated glucose derivatives from Astragalus bhotanensis

Four previously undescribed hydroxypropionylated d-glucose derivatives, astrabhotins A–D (1–4), along with ten known compounds α-d-glucose (5), β-d-glucose (6), quebrachitol (7), 3-hydroxypropionic acid (8), oleic acid (9), isoliquiritigenin (10), liquiritigenin (11), odoratin (12), 7β-hydroxysitosterol (13) and daucosterol (14), were isolated from the roots of Astragalus bhotanensis. Their structures were elucidated based on the analyses of extensive spectroscopic data and physicochemical properties. Astrabhotin A (1) reduced the writhing response remarkably with 52.5% inhibition by acetic acid induced writhing test. The analgesic effect of 1 was stronger than the standard drug aspirin. In addition, compounds 1 and 3 showed significant antioxidant activities with IC50 values of 9.9 ± 0.2 and 7.9 ± 0.4 μg/mL, and exhibited weak or moderate cytotoxicity against HepG2 cells with IC50 values of 106.6 ± 2.7 and 42.0 ± 0.9 μg/mL, respectively.

Role of root exudates on assimilation of phosphorus in young and old Arabidopsis thaliana plants.

The role of root exudates has long been recognized for its potential to improve nutrient use efficiency in cropping systems. However, studies addressing the variability of root exudates involved in phosphorus solubilization across plant developmental stages remain scarce. Here, we grew Arabidopsis thaliana seedlings in sterile liquid culture with a low, medium, or high concentration of phosphate and measured the composition of the root exudate at seedling, vegetative, and bolting stages. The exudates changed in response to the incremental addition of phosphorus, starting from the vegetative stage. Specific metabolites decreased in relation to phosphate concentration supplementation at specific stages of development. Some of those metabolites were tested for their phosphate solubilizing activity, and 3-hydroxypropionic acid, malic acid, and nicotinic acid were able to solubilize calcium phosphate from both solid and liquid media. In summary, our data suggest that plants can release distinct compounds to deal with phosphorus deficiency needs influenced by the phosphorus nutritional status at varying developmental stages.

Development of orthogonal T7 expression system in Klebsiella pneumoniae.

Most expression systems are tailored for model organisms rather than nonmodel organisms. However, heterologous gene expression in model organisms constrains the innate advantages of original strain carrying gene of interest. In this study, T7 expression system was developed in nonmodel bacterium Klebsiella pneumoniae for production of chemicals. First, we engineered a recombinant K. pneumoniae strain harboring two vectors. One vector was used to express T7 RNA polymerase (T7 RNAP) which would drive the expression of egfp in the other vector. This recombinant strain demonstrated 15.73-fold of fluorescence relative to wild-type K. pneumoniae and showed similar level of fluorescence to recombinant Escherichia coli overexpressing egfp. When egfp was replaced by puuC, an endogenous aldehyde dehydrogenase catalyzing 3-hydroxypropionic acid (3-HP) biosynthesis in K. pneumoniae, the recombinant strain coexpressing T7 RNAP and PuuC showed high-level PuuC expression. In shake-flask cultivation, this recombinant strain produced 1.72 g/L 3-HP in 24 hr, which was 3.24 times that of wild-type K. pneumoniae (0.53 g/L). To mitigate plasmid burden, the vector expressing T7 RNAP was eliminated, but the T7 RNAP expression cassette was integrated into K. pneumoniae genome. The resulting strain harboring only PuuC expression vector produced 2.44 g/L 3-HP in 24 hr under shake-flask conditions, which was 1.46 times that of the strain harboring both T7 RNAP and PuuC expression vectors. In bioreactor cultivation, this strain generated 67.59 g/L 3-HP and did not show significantly halted growth. Overall, these results indicate that the engineered T7 expression system functioned efficiently in K. pneumoniae. This study provides a paradigm for the development of T7 expression system in prokaryotes.

Biochemical and anaplerotic applications of in vitro models of propionic acidemia and methylmalonic acidemia using patient-derived primary hepatocytes

Propionic acidemia (PA) and methylmalonic acidemia (MMA) are autosomal recessive disorders of propionyl-CoA (P-CoA) catabolism, which are caused by a deficiency in the enzyme propionyl-CoA carboxylase or the enzyme methylmalonyl-CoA (MM-CoA) mutase, respectively. The functional consequence of PA or MMA is the inability to catabolize P-CoA to MM-CoA or MM-CoA to succinyl-CoA, resulting in the accumulation of P-CoA and other metabolic intermediates, such as propionylcarnitine (C3), 3-hydroxypropionic acid, methylcitric acid (MCA), and methylmalonic acid (only in MMA). P-CoA and its metabolic intermediates, at high concentrations found in PA and MMA, inhibit enzymes in the first steps of the urea cycle as well as enzymes in the tricarboxylic acid (TCA) cycle, causing a reduction in mitochondrial energy production. We previously showed that metabolic defects of PA could be recapitulated using PA patient-derived primary hepatocytes in a novel organotypic system. Here, we sought to investigate whether treatment of normal human primary hepatocytes with propionate would recapitulate some of the biochemical features of PA and MMA in the same platform. We found that high levels of propionate resulted in high levels of intracellular P-CoA in normal hepatocytes. Analysis of TCA cycle intermediates by GC–MS/MS indicated that propionate may inhibit enzymes of the TCA cycle as shown in PA, but is also incorporated in the TCA cycle, which does not occur in PA. To better recapitulate the disease phenotype, we obtained hepatocytes derived from livers of PA and MMA patients. We characterized the PA and MMA donors by measuring key proximal biomarkers, including P-CoA, MM-CoA, as well as clinical biomarkers propionylcarnitine-to-acetylcarnitine ratios (C3/C2), MCA, and methylmalonic acid. Additionally, we used isotopically-labeled amino acids to investigate the contribution of relevant amino acids to production of P-CoA in models of metabolic stability or acute metabolic crisis. As observed clinically, we demonstrated that the isoleucine and valine catabolism pathways are the greatest sources of P-CoA in PA and MMA donor cells and that each donor showed differential sensitivity to isoleucine and valine. We also studied the effects of disodium citrate, an anaplerotic therapy, which resulted in a significant increase in the absolute concentration of TCA cycle intermediates, which is in agreement with the benefit observed clinically. Our human cell-based PA and MMA disease models can inform preclinical drug discovery and development where mouse models of these diseases are inaccurate, particularly in well-described species differences in branched-chain amino acid catabolism.

Construction and analysis of an artificial consortium based on the fast-growing cyanobacterium Synechococcus elongatus UTEX 2973 to produce the platform chemical 3-hydroxypropionic acid from CO2

BackgroundCyanobacterial carbohydrates, such as sucrose, have been considered as potential renewable feedstock to support the production of fuels and chemicals. However, the separation and purification processes of these carbohydrates will increase the production cost of chemicals. Co-culture fermentation has been proposed as an efficient and economical way to utilize these cyanobacterial carbohydrates. However, studies on the application of co-culture systems to achieve green biosynthesis of platform chemicals are still rare.ResultsIn this study, we successfully achieved one-step conversion of sucrose derived from cyanobacteria to fine chemicals by constructing a microbial consortium consisting of the fast-growing cyanobacterium Synechococcus elongatus UTEX 2973 and Escherichia coli to sequentially produce sucrose and then the platform chemical 3-hydroxypropionic acid (3-HP) from CO2 under photoautotrophic growth conditions. First, efforts were made to overexpress the sucrose permease-coding gene cscB under the strong promoter Pcpc560 in S. elongatus UTEX 2973 for efficient sucrose secretion. Second, the sucrose catabolic pathway and malonyl-CoA-dependent 3-HP biosynthetic pathway were introduced into E. coli BL21 (DE3) for heterologous biosynthesis of 3-HP from sucrose. By optimizing the cultivation temperature from 37 to 30 °C, a stable artificial consortium system was constructed with the capability of producing 3-HP at up to 68.29 mg/L directly from CO2. In addition, cell growth of S. elongatus UTEX 2973 in the consortium was enhanced, probably due to the quick quenching of reactive oxygen species (ROS) in the system by E. coli, which in turn improved the photosynthesis of cyanobacteria.ConclusionThe study demonstrated the feasibility of the one-step conversion of sucrose to fine chemicals using an artificial consortium system. The study also confirmed that heterotrophic bacteria could promote the cell growth of cyanobacteria by relieving oxidative stress in this microbial consortium, which further suggests the potential value of this system for future industrial applications.

High-level production of 3-hydroxypropionic acid from glycerol as a sole carbon source using metabolically engineered Escherichia coli.

As climate change is an important environmental issue, the conventional petrochemical-based processes to produce valuable chemicals are being shifted toward eco-friendly biological-based processes. In this study, 3-hydroxypropionic acid (3-HP), an industrially important three carbon (C3) chemical, was overproduced by metabolically engineered Escherichia coli using glycerol as a sole carbon source. As the first step to construct a glycerol-dependent 3-HP biosynthetic pathway, the dhaB1234 and gdrAB genes from Klebsiella pneumoniae encoding glycerol dehydratase and glycerol reactivase, respectively, were introduced into E. coli to convert glycerol into 3-hydroxypropionaldehyde (3-HPA). In addition, the ydcW gene from K. pneumoniae encoding γ-aminobutyraldehyde dehydrogenase, among five aldehyde dehydrogenases examined, was selected to further convert 3-HPA to 3-HP. Increasing the expression level of the ydcW gene enhanced 3-HP production titer and reduced 1,3-propanediol production. To enhance 3-HP production, fed-batch fermentation conditions were optimized by controlling dissolved oxygen (DO) level and employing different feeding strategies including intermittent feeding, pH-stat feeding, and continuous feeding strategies. Fed-batch culture of the final engineered E. coli strain with DO control and continuous feeding strategy produced 76.2 g/L of 3-HP with the yield and productivity of 0.457 g/g glycerol and 1.89 g·L-1 ·h-1 , respectively. To the best of our knowledge, this is the highest 3-HP productivity achieved by any microorganism reported to date.

Use of acetate for the production of 3-hydroxypropionic acid by metabolically-engineered Pseudomonas denitrificans

The use of acetate as carbon feedstock can enhance sustainability and economics of the current bio-productions. This study explored the potential of acetate for the production of 3-hydroxypropionic acid by engineered Pseudomonas denitrificans. Heterologous mcr (encoding malonyl-CoA reductase) from Chloroflexus aurantiacus and endogenous accABCD (encoding acetyl-CoA carboxylase) were overexpressed in P. denitrificans. Carbon flux to 3-HP synthesis at the malonyl-CoA node was promoted by suppressing fatty acid synthesis through addition of cerulenin or deletion of fabF gene. In addition, stimulation of glyoxylate shunt and/or TCA cycle were attempted. Recombinant P. denitrificans overexpressing mcr and accABCD produced 19.3 mM 3-HP with cerulenin addition, and 14.2 mM with fabF deletion, respectively. Furthermore, the non-growing cells devoid of fabF could continuously produce 3-HP up to 40.4 mM without losing its production activity for 22 h. This study demonstrates that acetate is a good substrate for 3-HP production by recombinant P. denitrificans.

Carglumic Acid Contributes to a Favorable Clinical Course in a Case of Severe Propionic Acidemia

Propionic acidemia (PA) is manifested as an abnormal accumulation of propionic acid and its metabolites, including methylcitrate, 3-hydroxypropionic acid, and propionylglycine, and is caused by a defect of propionyl-CoA carboxylase. PA is complicated by acute life-threatening metabolic crises, which are precipitated by a catabolic state and result in multiple organ failure or even death, if untreated. A neonate with PA recovered from the first metabolic crisis 3 days after birth but developed a second metabolic crisis during the recovery phase. This patient was considered to have severe PA and was accordingly given carglumic acid treatment in combination with carnitine supplementation and protein restriction, which was expected to prevent a recurrent metabolic attack. The patient did not develop hyperammonemia after receiving carglumic acid and was never hospitalized. Moreover, she did not present with acidosis even during viral infection. At 26 months of age, she led a stable life while receiving carglumic acid and regular rehabilitation. Carglumic acid treatment in combination with carnitine supplementation and protein restriction prevented metabolic decompensation, which would have otherwise required hospitalization, and resulted in improved quality of life and developmental outcomes.

An Asymptotic Analysis of the Malonyl-CoA Route to 3-Hydroxypropionic Acid in Genetically Engineered Microbes

There has been recent interest in creating an efficient microbial production route for 3-hydroxypropionic acid, an important platform chemical. We develop and solve a mathematical model for the time-dependent metabolite concentrations in the malonyl-CoA pathway for 3-hydroxypropionic acid production in microbes, using a combination of numerical and asymptotic methods. This allows us to identify the most important targets for enzyme regulation therein under conditions of plentiful and sparse pyruvate, and to quantify their relative importance. In our model, we account for sinks of acetyl-CoA and malonyl-CoA to, for example, the citric acid cycle and fatty acid biosynthesis, respectively. Notably, in the plentiful pyruvate case we determine that there is a bifurcation in the asymptotic structure of the system, the crossing of which corresponds to a significant increase in 3-hydroxypropionic acid production. Moreover, we deduce that the most significant increases to 3-hydroxypropionic acid production can be obtained by up-regulating two specific enzymes in tandem, as the inherent nonlinearity of the system means that a solo up-regulation of either does not result in large increases in production. The types of issue arising here are prevalent in synthetic biology applications, and it is hoped that the system considered provides an instructive exemplar for broader applications.

Algicidal activity recovery by a Li-doped up-conversion material converting visible light into UV

Citrobacter sp. R1, a highly efficient algicidal bacterium, undergoes a serious degeneration after long-term preservation. Here, we prepared a Li-doped up-conversion material ([Y2SiO5; Li (1%)]) to recover the degenerated Strain R1. As a result, the [Y2SiO5; Li (1%)] successfully enhanced the maximum algicidal activity of Strain R1 from 43.2 ± 1.2 to 72.5 ± 1.3%. In terms of recovery mechanisms, 1) the [Y2SiO5; Li (1%)] could convert visible light into UVC and UVA with unusual wavelengths, creating conditions for inducing reverse mutation in Strain R1, affecting the metabolism of Strain R1 and then recovering the bacterium; 2) the [Y2SiO5; Li (1%)] did not only enhance the secretion of 3-hydroxypropionic acid, the algicidal compound which Strain R1 continues to produce even after degeneration, but also it recovered the ability of Strain R1 to secrete three new algicidal compounds. This study provides a new idea of employing up-conversion material [Y2SiO5; Li (1%)] in microbial degeneration recovery.

New insights into cheddar cheese microbiota-metabolome relationships revealed by integrative analysis of multi-omics data

Cheese microbiota and metabolites and their inter-relationships that underpin specific cheese quality attributes remain poorly understood. Here we report that multi-omics and integrative data analysis (multiple co-inertia analysis, MCIA) can be used to gain deeper insights into these relationships and identify microbiota and metabolite fingerprints that could be used to monitor product quality and authenticity. Our study into different brands of artisanal and industrial cheddar cheeses showed that Streptococcus, Lactococcus and Lactobacillus were the dominant taxa with overall microbial community structures differing not only between industrial and artisanal cheeses but also among different cheese brands. Metabolome analysis also revealed qualitative and semi-quantitative differences in metabolites between different cheeses. This also included the presence of two compounds (3-hydroxy propanoic acid and O-methoxycatechol-O-sulphate) in artisanal cheese that have not been previously reported in any type of cheese. Integrative analysis of multi-omics datasets revealed that highly similar cheeses, identical in age and appearance, could be distinctively clustered according to cheese type and brand. Furthermore, the analysis detected strong relationships, some previously unknown, which existed between the cheese microbiota and metabolome, and uncovered specific taxa and metabolites that contributed to these relationships. These results highlight the potential of this approach for identifying product specific microbe/metabolite signatures that could be used to monitor and control cheese quality and product authenticity.