Acetate Supplementation Research Articles

A High-Fiber Diet or Dietary Supplementation of Acetate Attenuate Hyperoxia-Induced Acute Lung Injury.

A high fiber diet (HFD) and dietary supplementation with acetate have been reported to have beneficial effects in a variety of diseases. We investigated the effects of a HFD and acetate supplementation on the gut microbiota and hyperoxia-induced acute lung injury (HALI) in mice. Mice were fed a control diet, HFD, or acetate supplementation for three weeks, and their gut microbiome composition, lung tissues, and bronchoalveolar lavage fluid (BALF) were examined after exposure to ambient air or hyperoxia. Both the HFD and acetate supplementation modified the gut microbiota community and increased the proportion of acetate-producing bacteria in mice exposed to hyperoxia. The HFD and acetate supplementation also increased the abundance of Bacteroides acidifaciens and reduced gut dysbiosis according to the ratio of Firmicutes to Bacteroidetes. Compared with hyperoxia-exposed mice fed a control diet, both the HFD and acetate supplementation significantly increased the survival time while reducing the severity of pulmonary edema and the concentrations of protein and inflammatory mediators in BALF. Moreover, the HFD and acetate supplementation reduced the production of free radicals, attenuated NF-κB signaling activation, and decreased apoptosis in the lung tissues. Overall, this study indicates that a HFD or acetate supplementation reduces the severity of HALI through alterations in the gut microbiota to exert anti-inflammatory effects.

Dietary sodium acetate improves high-fat diet utilization through promoting differential nutrients metabolism between liver and muscle in Nile tilapia (Oreochromis niloticus)

High-fat diet (HFD) often causes many negative effects and impairs fish growth. Short-chain fatty acids (SCFAs) such as acetates modify metabolic disorders and regulate body homeostasis. However, the effects of sodium acetate on alleviating HFD in fish is currently unknown. This study investigated the role of using dietary sodium acetate to alleviate adverse effects caused by HFD in fish. Three replicates (thirty fish each) of 4.8 ± 0.30 g Nile tilapia (Oreochromis niloticus) were fed with control diet (Con), high-fat diet (HFD) or HFD containing sodium acetate diet (HFD + NaAc) for eight weeks. After the feeding trial, Nile tilapia fed with HFD increased significantly tissue lipid deposition, reduced insulin sensitivity and suppressed glucose and lipid metabolism in both liver and muscle, accompanied with signs of metabolic disorders and liver damage. Moreover, HFD feeding inhibited muscle protein synthesis and impaired fish growth performance. However, the fish fed on HFD + NaAc improved significantly oxidative stress, inflammation, apoptosis and injury in liver compared to those fed on HFD. More importantly, dietary sodium acetate supplementation enhanced insulin sensitivity, promoted glucose catabolic utilization and protein synthesis in muscle through activation of insulin and mTOR signaling pathways, respectively, and markedly improved the growth performance. In contrast, dietary sodium acetate promoted hepatic pentose phosphate pathway, hepatic glycogen accumulation, and activated lipid catabolism to alleviate hepatic lipid deposition. Our study illustrates that sodium acetate alleviates differentially the adverse effects induced by feeding Nile tilapia with HFD in muscle and liver. SCFAs such as acetate may be used for improving HFD utilization in fish nutrition.

Metabolic changes of the acetogen Clostridium sp. AWRP through adaptation to acetate challenge.

In this study, we report the phenotypic changes that occurred in the acetogenic bacterium Clostridium sp. AWRP as a result of an adaptive laboratory evolution (ALE) under the acetate challenge. Acetate-adapted strain 46 T-a displayed acetate tolerance to acetate up to 10 g L-1 and increased ethanol production in small-scale cultures. The adapted strain showed a higher cell density than AWRP even without exogenous acetate supplementation. 46 T-a was shown to have reduced gas consumption rate and metabolite production. It was intriguing to note that 46 T-a, unlike AWRP, continued to consume H2 at low CO2 levels. Genome sequencing revealed that the adapted strain harbored three point mutations in the genes encoding an electron-bifurcating hydrogenase (Hyt) crucial for autotrophic growth in CO2 + H2, in addition to one in the dnaK gene. Transcriptome analysis revealed that most genes involved in the CO2-fixation Wood-Ljungdahl pathway and auxiliary pathways for energy conservation (e.g., Rnf complex, Nfn, etc.) were significantly down-regulated in 46 T-a. Several metabolic pathways involved in dissimilation of nucleosides and carbohydrates were significantly up-regulated in 46 T-a, indicating that 46 T-a evolved to utilize organic substrates rather than CO2 + H2. Further investigation into degeneration in carbon fixation of the acetate-adapted strain will provide practical implications for CO2 + H2 fermentation using acetogenic bacteria for long-term continuous fermentation.

The effect of gossypol acetate on the fertility of boars


 This study evaluated the effect of gossypol acetate, a potential antifertility compound, on semen quality and testicular histopathology of boars. Six boars were allotted into two experimental groups. Half of the animals were fed a diet that was supplemented with 2 mg of gossypol acetate per kg body weight per day for 9 weeks, control animals received a gossypol acetate-free feed. During the experimental period, semen was collected weekly from each boar and semen parameters were recorded. The animals were then euthanized, testicular samples were collected and histopathological examination of the testicular cells, as well as morphometrical analysis of the seminiferous tubules, were performed. The percentage of spermatozoa showing tail abnormalities increased significantly (P=0.017) in the semen of boars fed gossypol acetate-supplemented feed, while several other semen parameters deteriorated without showing statistical significance. Gossypol acetate supplementation also led to a decrease (P=0.042) in the number of spermatogonia in the seminiferous tubules and an increase (P=0.020) in the number of vacuoles in the seminiferous epithelium, consistent with changes seen in cases of male reproductive toxicity. In conclusion, gossypol acetate negatively affected a number of semen characteristics and also had detrimental effects on the histopathology of the testes.

HEAT SHOCK PROTEINS AS BIOMARKERS OF FISH POLLUTION

Нeat Shock Proteins (HSP) belong to the natural biomarkers, which are important indicators for animal diseases diagnostics and / or instrument of analyzing the effects on organism of the habitat deteriorating factors. The contamination of water by heavy metals has adverse effect on fish organism. Even in a small quantity, such heavy metal as lead is very dangerous. The analysis of toxic effects of the lead ions on the level of expression of such heat shock proteins as HSP60, HSP70 and HSC70 family in leukocytes, liver, brain and gills of the scaly carp was the main goal of our investigation.
 During 96 hours the fish were kept in the aqueous environment of a tank which additionally was supplemented with Pb (CH3COO)2. The control group of fish was maintained for the similar period of time under the same conditions, without lead acetate supplementation.
 Concentration of HSP60, HSP70 was determinated by the dot-blot-analysis due to application of monoclonal antibodies against heat shock proteins SAB4501464 (Sigma, USA), [5A5] (ab2787) (Abcam, USA) and [1B5] (ab19136) (Abcam, USA). Detection of immune complexes was performed by using the commercial substrate solution for alkaline phosphatase - CDP-Star (Tropix, UK). Visualization was done by using X-ray film ECL HyperFilm (Amersham, USA) and a kit for films developing (Kodak). The images were processed using the software package GelPro (Version 3.1, USA).
 The significant dose-depended increase (p<0,001) in all experimental groups of HSP60 and HSP70 concentrations in leukocytes, liver, gill and brain has been detected, applying dot-blot analysis. At the same time, significant changes in expression of HSC70 protein have not been established. It is the evidence that stress-proteins are the sensitive markers of toxic effects of excessive concentration of lead.

Regulation of acidogenic fermentation through exogenous additives for promoting carbon conversion of food waste in two-phase anaerobic system

In this study, exogenous Megasphaera elsdenii inoculum and acetate supplementation were introduced at the acidogenic phase to regulate the acidogenic fermentation pathway and assess their effects on food waste (FW) carbon conversion in two-phase anaerobic digestion (AD) system. These two additives significantly accelerated organic removal efficiency and subsequently increased FW hydrolysis and acidogenesis by 16% and 35%, respectively. As expected, two exogenous additives promoted butyrate fermentation during FW acidogenesis. With regard to the role of exogenous additives, both hydrogen and butyrate yields increased by over 60%. This desired increment resulted in a 25% increase in methane production. The overall carbon conversion from FW in the integrated two-phase AD system was enhanced by biochemical additives, which was 1.3-fold higher than that in control without any additives. Collectively, findings demonstrate the feasibility of regulating acidogenic fermentation via exogenous biochemical additives and its benefits on FW carbon conversion during AD process.

Effects of supplementation of sodium acetate on rumen fermentation and microbiota in postpartum dairy cows.

The primary product of rumen fermentation is acetic acid, and its sodium salt is an excellent energy source for post-partum cows to manage negative energy balance (NEB). However, it is unknown how adding sodium acetate (NAc) may affect the rumen bacterial population of post-partum cows. Using the identical nutritional total mixed ration (TMR), this research sought to characterize the impact of NAc supplementation on rumen fermentation and the composition of bacterial communities in post-partum cows. After calving, 24 cows were randomly assigned to two groups of 12 cows each: a control group (CON) and a NAc group (ACE). All cows were fed the same basal TMR with 468 g/d NaCl added to the TMR for the CON group and 656 g/d NAc added to the TMR for the ACE group for 21 days after calving. Ruminal fluid was collected before morning feeding on the last day of the feeding period and analyzed for rumen bacterial community composition by 16S rRNA gene sequencing. Under the identical TMR diet conditions, NAc supplementation did not change rumen pH but increased ammonia nitrogen (NH3-N) levels and microbial crude protein (MCP) concentrations. The administration of NAc to the feed upregulated rumen concentrations of total volatile fatty acids (TVFA), acetic, propionic, isovaleric and isobutyric acids without affecting the molar ratio of VFAs. In the two experimental groups, the Bacteroidota, Firmicutes, Patescibacteria and Proteobacteria were the dominant rumen phylum, and Prevotella was the dominant rumen genus. The administration of NAc had no significant influence on the α-diversity of the rumen bacterial community but upregulated the relative abundance of Prevotella and downregulated the relative abundance of RF39 and Clostridia_UCG_014. In conclusion, the NAc supplementation in the post-peripartum period altered rumen flora structure and thus improved rumen fermentation in dairy cows. Our findings provide a reference for the addition of sodium acetate to alleviate NEB in cows during the late perinatal period.

Acetate supplementation restores cognitive deficits caused by ARID1A haploinsufficiency in excitatory neurons

Mutations in AT‐rich interactive domain‐containing protein 1A (ARID1A) cause Coffin‐Siris syndrome (CSS), a rare genetic disorder that results in mild to severe intellectual disabilities. However, the biological role of ARID1A in the brain remains unclear. In this study, we report that the haploinsufficiency of ARID1A in excitatory neurons causes cognitive impairment and defects in hippocampal synaptic transmission and dendritic morphology in mice. Similarly, human embryonic stem cell‐derived excitatory neurons with deleted ARID1A exhibit fewer dendritic branches and spines, and abnormal electrophysiological activity. Importantly, supplementation of acetate, an epigenetic metabolite, can ameliorate the morphological and electrophysiological deficits observed in mice with Arid1a haploinsufficiency, as well as in ARID1A‐null human excitatory neurons. Mechanistically, transcriptomic and ChIP‐seq analyses demonstrate that acetate supplementation can increase the levels of H3K27 acetylation at the promoters of key regulatory genes associated with neural development and synaptic transmission. Collectively, these findings support the essential roles of ARID1A in the excitatory neurons and cognition and suggest that acetate supplementation could be a potential therapeutic intervention for CSS.

Interaction of sodium acetate supplementation and dietary fiber level on feeding behavior, digestibility, milk synthesis, and plasma metabolites.

Acetate supplementation has been shown to increase milk fat yield in diets with low risk of biohydrogenation-induced milk fat depression. The interaction of acetate supplementation with specific dietary factors that modify rumen fermentation and short-chain fatty acid (FA) synthesis has not been investigated. The objective of this experiment was to determine the effect of acetate supplemented as sodium acetate at 2 dietary fiber levels. Our hypothesis was that acetate would increase milk fat production more in animals fed the low-fiber diet. Twelve lactating multiparous Holstein cows were arranged in a 4 × 4 Latin square design balanced for carryover effects with a 2 × 2 factorial arrangement of dietary fiber level and acetate supplementation with 21-d experimental periods. The high-fiber diet had 32% neutral detergent fiber and 21.8% starch, and the low-fiber diet had 29.5% neutral detergent fiber and 28.7% starch created by substitution of forages predominantly for ground corn grain. Acetate was supplemented in the diet at an average 2.8% of dry matter (DM) to provide approximately 10 mol/d of acetate as anhydrous sodium acetate. Acetate supplementation increased DM intake by 6%, with no effect on meal frequency or size. Furthermore, acetate supplementation slightly increased total-tract apparent DM digestibility and tended to increase organic matter digestibility. Acetate supplementation increased milk fat concentration and yield by 8.6 and 10.5%, respectively, but there was no interaction with dietary fiber. The increase in milk fat synthesis was associated with 46 and 85 g/d increases in the yield of de novo (<16C) and mixed source (16C) FA, respectively, with no changes in yield of preformed FA (>16C). There was a 9% increase in the concentration of milk mixed-source FA and a 7% decrease in milk preformed FA with acetate supplementation, regardless of dietary fiber level. Acetate supplementation also increased the concentrations of plasma acetate and β-hydroxybutyrate, major metabolic substrates for mammary lipogenesis. Overall, acetate supplementation increased milk fat yield regardless of dietary fiber level through an increase mostly caused by an increase in longer-chain de novo FA, suggesting stimulation of mammary lipogenesis. The heightened mammary de novo lipogenesis was supported by an increase in the concentration of metabolic substrates in plasma.

Vitamins and their derivatives synergistically promote hair shaft elongation ex vivo via PlGF/VEGFR-1 signalling activation

BackgroundAlthough vitamins or their derivatives (Vits), such as panthenyl ethyl ether, tocopherol acetate, and pyridoxine, have been widely used in topical hair care products, their efficacy and mode of action have been insufficiently studied. ObjectiveTo elucidate the biological influence of Vits on hair follicles and determine the underlying mechanisms. MethodsA mouse vibrissa hair follicle organ culture model was utilized to evaluate the effects of Vits on hair shaft elongation. Gene and protein expression analyses and histological investigations were conducted to elucidate the responsible mechanisms. A human hair follicle cell culture was used to assess the clinical relevance. ResultsIn organ culture models, the combination of panthenyl ethyl ether, tocopherol acetate, and pyridoxine (namely, PPT) supplementation significantly promoted hair shaft elongation. PPT treatment enhanced hair matrix cell proliferation by 1.9-fold compared to controls, as demonstrated by Ki67-positive immunoreactivity. PPT-treated mouse dermal papillae exhibited upregulated Placental growth factor (Plgf) by 1.6-fold compared to controls. Importantly, the addition of PlGF neutralizing antibodies to the ex vivo culture diminished the promotive effect on hair growth and increase in VEGFR-1 phosphorylation achieved by PPT. A VEGFR-1 inhibitor also inhibited the promotion of hair growth. Microarray analysis suggested synergistic summation of individual Vits’ bioactivity, putatively explaining the effect of PPT. Moreover, PPT increased PlGF secretion in cultured human dermal papilla cells. ConclusionOur findings suggested that PPT promoted hair shaft elongation by activating PlGF/VEGFR-1 signalling. The current study can shed light on the previously underrepresented advantage of utilizing Vits in hair care products.

Antitumor Effect of Zinc Acetate in Hepatocellular Carcinoma Cell Lines via the Induction of Apoptosis.

We aimed to verify antitumor effects of zinc acetate on hepatocellular carcinoma (HCC) in vitro. Five HCC cell lines (HepG2, Hep3B, Huh7, HLE and Alex) were used to evaluate the antitumor effects of zinc acetate. Cell viability was determined by the Cell Counting Kit-8 assay. The cell-cycle alteration was evaluated by a flow cytometric analysis and the detection of cell cycle-related proteins. Apoptosis was determined based on the caspase-cleaved cytokeratin 18 (cCK18) levels. The microRNAs (miRNAs) related to an antitumor effect of zinc acetate were identified using microarrays. Zinc acetate significantly inhibited the proliferation of HCC cells in a dose-dependent manner. The treatment with zinc acetate resulted in significantly increased cCK18 levels in the supernatant and enhanced the expression of heme oxygenase-1 (HO-1) in HCC cells. The flow cytometric analysis revealed an increase of HCC cells in the S and G2/M phases by the administration of zinc acetate, and the expressions of Cdk2 and cyclin E were increased. The miRNA expression profile of the HCC cells treated with zinc acetate was extremely different from that of the untreated HCC cells. These results suggest that the zinc acetate supplementation induces the apoptosis of HCC cells, but does not affect the cell cycle progression. Upregulation of HO-1 and the alteration of miRNAs' profile may be involved in antitumor effects of zinc acetate in HCC cells.

Inulin-type fructans change the gut microbiota and prevent the development of diabetic nephropathy

Diabetic nephropathy (DN) is the most common cause of end-stage renal disease, and few treatment options that prevent the progressive loss of renal function are available. Studies have shown that dietary fiber intake improves kidney diseases and metabolism-related diseases, most likely through short-chain fatty acids (SCFAs). The present study aimed to examine the protective effects of inulin-type fructans (ITFs) on DN through 16 S rRNA gene sequencing, gas chromatographymass spectrometry (GCMS) analysis and fecal microbiota transplantation (FMT). The results showed that ITFs supplementation protected against kidney damage in db/db mice and regulated the composition of the gut microbiota. Antibiotic treatment and FMT experiments further demonstrated a key role of the gut microbiota in mediating the beneficial effects of ITFs. The ITFs treatment-induced changes in the gut microbiota led to an enrichment of SCFA-producing bacteria, especially the genera Akkermansia and Candidatus Saccharimonas, which increased the fecal and serum acetate concentrations. Subsequently, acetate supplementation improved glomerular damage and renal fibrosis by attenuating mitochondrial dysfunction and reducing toxic glucose metabolite levels. In conclusion, ITFs play a renoprotective role by modulating the gut microbiota and increasing acetate production. Furthermore, acetate mediates renal protection by regulating glucose metabolism, decreasing glycotoxic product levels and improving mitochondrial function.

Maternal Acetate Supplementation Reverses Blood Pressure Increase in Male Offspring Induced by Exposure to Minocycline during Pregnancy and Lactation.

Emerging evidence supports that hypertension can be programmed or reprogrammed by maternal nutrition. Maternal exposures during pregnancy, such as maternal nutrition or antibiotic use, could alter the offspring’s gut microbiota. Short-chain fatty acids (SCFAs) are the major gut microbiota-derived metabolites. Acetate, the most dominant SCFA, has shown its antihypertensive effect. Limited information exists regarding whether maternal acetate supplementation can prevent maternal minocycline-induced hypertension in adult offspring. We exposed pregnant Sprague Dawley rats to normal diet (ND), minocycline (MI, 50 mg/kg/day), magnesium acetate (AC, 200 mmol/L in drinking water), and MI + AC from gestation to lactation period. At 12 weeks of age, four groups (n = 8/group) of male progeny were sacrificed. Maternal acetate supplementation protected adult offspring against minocycline-induced hypertension. Minocycline administration reduced plasma acetic acid level, which maternal acetate supplementation prevented. Additionally, acetate supplementation increased the protein level of SCFA receptor G protein-coupled receptor 41 in the offspring kidneys. Further, minocycline administration and acetate supplementation significantly altered gut microbiota composition. Maternal acetate supplementation protected minocycline-induced hypertension accompanying by the increases in genera Roseburia, Bifidobacterium, and Coprococcus. In sum, our results cast new light on targeting gut microbial metabolites as early interventions to prevent the development of hypertension, which could help alleviate the global burden of hypertension.

Electron donor addition for stimulating the microbial degradation of 1,4 dioxane by sequential batch membrane bioreactor: A techno-economic approach

The presence of 1,4 dioxane in wastewater is associated with severe health and environmental issues. The removal of this toxic contaminant from the industrial effluents prior to final disposal is necessary. The study comprehensively evaluates the performance of sequential batch membrane bioreactor (MBR) for treating wastewater laden with 1,4 dioxane. Acetate was supplemented to the wastewater feed as an electron donor for enhancing and stimulating the microbial growing activities towards the degradation of 1,4 dioxane. The removal efficiency of 1,4 dioxane was maximized to 87.5 ± 6.8% using an acetate to dioxane (A/D) ratio of 4.0, which was substantially dropped to 31.06 ± 3.7% without acetate addition. Ethylene glycol, glyoxylic acid, glycolic acid, and oxalic acid were the main metabolites of 1,4 dioxane biodegradation using mixed culture bacteria. The 1,4 dioxane degrading bacteria, particularly the genus of Acinetobacter, were promoted to 92% at the A/D ratio of 4.0. This condition encouraged as well the increase of the main 1,4 dioxane degraders, i.e., Xanthomonadales (12.5%) and Pseudomonadales (9.1%). However, 50% of the Sphingobacteriales and 82.5% of Planctomycetes were reduced due to the inhibition effect of the 1,4 dioxane contaminate. Similarly, the relative abundance of Firmicutes, Verrucomicrobia, Chlamydiae, Actinobacteria, Chloroflexi, and Nitrospirae was reduced in the MBR at the A/D ratio of 4.0. The results derived from the microbial analysis and metabolites detection at different A/D ratios indicated that acetate supplementation (as an electron donor) maintained an essential role in encouraging the microorganisms to produce the monooxygenase enzymes responsible for the biodegradation process. Economic feasibility of such a MBR system showed that for a designed flow rate of 30 m3∙d−1, the payback period from reusing the treated wastewater would reach 6.6 yr. The results strongly recommend the utilization of mixed culture bacteria growing on acetate for removing 1,4 dioxane from the wastewater industry, achieving dual environmental and economic benefits.

Metabolic engineering of oleaginous yeast Rhodotorula toruloides for overproduction of triacetic acid lactone

The plant‐sourced polyketide triacetic acid lactone (TAL) has been recognized as a promising platform chemical for the biorefinery industry. However, its practical application was rather limited due to low natural abundance and inefficient cell factories for biosynthesis. Here, we report the metabolic engineering of oleaginous yeast Rhodotorula toruloides for TAL overproduction. We first introduced a 2‐pyrone synthase gene from Gerbera hybrida (GhPS) into R. toruloides and investigated the effects of different carbon sources on TAL production. We then systematically employed a variety of metabolic engineering strategies to increase the flux of acetyl‐CoA by enhancing its biosynthetic pathways and disrupting its competing pathways. We found that overexpression of ATP‐citrate lyase (ACL1) improved TAL production by 45% compared to the GhPS overexpressing strain, and additional overexpression of acetyl‐CoA carboxylase (ACC1) further increased TAL production by 29%. Finally, we characterized the resulting strain I12‐ACL1‐ACC1 using fed‐batch bioreactor fermentation in glucose or oilcane juice medium with acetate supplementation and achieved a titer of 28 or 23 g/L TAL, respectively. This study demonstrates that R. toruloides is a promising host for the production of TAL and other acetyl‐CoA‐derived polyketides from low‐cost carbon sources.

Buffering culture solution significantly improves astaxanthin production efficiency of mixotrophic Haematococcus pluvialis

Sodium acetate (NaAc) supplementation, often used to increase the growth of H. pluvialis under low light, but promotes cell death under high light; its underlying reasons and solutions are rarely reported. Here, NaAc supplementation was found to rapidly increase pondus hydrogenii (pH) of culture solution, elevate reactive oxygen species (ROS), and cause cell death immediately under higher light. Adjusting pH of NaAc supplemented culture solution with 10 mM Tris-HCl once before high light significantly reduced cell mortality and increased astaxanthin yield. When verified in a 5-litre photobioreactor, this novel method produced over 4.0% of dry weight (DW) astaxanthin within only 8–10 days. In summary, this study explained reasons underlying NaAc supplementation-induced cell death and provided an rapid, easy and effective method to produce high amount of astaxanthin in H. pluvialis.

Lipotoxic Effects of Gossypol Acetate Supplementation on Hepatopancreas Fat Accumulation and Fatty Acid Profile in Cyprinus carpio

Cottonseed meal is an excellent protein source with low price and abundant nutrients, but the existence of free gossypol restricts its wide application in animal feeds. To explore the toxic effects of free gossypol, the effects of gossypol acetate on the growth performance, hepatopancreatic fat accumulation, and muscle fatty acid composition of the common carp (Cyprinus carpio) were investigated in the present study. Carp were randomly divided into four groups, which were fed a diet supplemented with 0, 170, 340, and 680 mg/kg gossypol acetate, respectively. After 60 days of feeding, the gossypol residue in the tissue, growth performance, blood biochemical indices, hepatopancreas histology, fatty acid components, and expression of key genes involved in lipid metabolism pathways were measured. It was found that the concentrations of free gossypol were increased significantly in the serum, bile, and hepatopancreas of carp fed 340 mg/kg and 680 mg/kg gossypol acetate ( P &lt; 0.05 ). Dietary gossypol acetate had a limited effect on growth performance, and high concentrations of gossypol acetate induced excessive hepatopancreatic fat deposition and hepatocyte injury in C. carpio. In addition, gossypol acetate decreased total polyunsaturated fatty acids (PUFAs) and docosahexaenoic acid in muscle. In the hepatopancreas, decreased HSL, PPARα, and CD36 may result in fat accumulation. In the muscle, decreased FADS2 may be the key reason for the decrease in PUFAs. Taken together, these findings indicated that dietary gossypol acetate can cause lipid metabolism disorders, such as hepatopancreatic damage and significant changes in tissue fatty acid profile, which are regulated by the key lipid metabolic genes in common carp.

Leptolyngbya sp. NIVA-CYA 255, a Promising Candidate for Poly(3-hydroxybutyrate) Production under Mixotrophic Deficiency Conditions.

Cyanobacteria are a promising source for the sustainable production of biodegradable bioplastics such as poly(3-hydroxybutyrate) (PHB). The auto-phototrophic biomass formation is based on light and CO2, which is an advantage compared to heterotrophic PHB-producing systems. So far, only a handful of cyanobacterial species suitable for the high-yield synthesis of PHB have been reported. In the present study, the PHB formation, biomass, and elemental composition of Leptolyngbya sp. NIVA-CYA 255 were investigated. Therefore, a three-stage cultivation process was applied, consisting of a growth stage; an N-, P-, and NP-depleted phototrophic stage; and a subsequent mixotrophic deficiency stage, initiated by sodium acetate supplementation. The extracted cyanobacterial PHB was confirmed by FTIR- and GC-MS analyses. Furthermore, the fluorescent dyes LipidGreen2 and Nile red were used for fluorescence-based monitoring and the visualization of PHB. LipidGreen2 was well suited for PHB quantification, while the application of Nile red was limited by fluorescence emission crosstalk with phycocyanin. The highest PHB yields were detected in NP- (325 mg g−1) and N-deficiency (213 mg g−1). The glycogen pool was reduced in all cultures during mixotrophy, while lipid composition was not affected. The highest glycogen yield was formed under N-deficiency (217 mg g−1). Due to the high carbon storage capacity and PHB formation, Leptolyngbya sp. NIVA-CYA 255 is a promising candidate for PHB production. Further work will focus on upscaling to a technical scale and monitoring the formation by LipidGreen2-based fluorometry.

A Nutrient-Based Cellular Model to Characterize Acetylation-Dependent Protein-Protein Interactions.

Cellular homeostasis requires the orderly expression of thousands of transcripts. Gene expression is regulated by numerous proteins that recognize post-translational modifications—in particular, the acetylation of lysine residues (Kac) on histones. In addition to affecting the general condensation state of the chromatin, acetylated histones act as anchor points for bromodomain (BRD)-containing adapter proteins. BRDs are the primary Kac reader domains in humans, and proteins containing them act as chromatin scaffolds that organize large networks of interactions to regulate transcription. To characterize BRD-dependent interaction networks, we established cell lines in which histone acetylation is dependent on acetate supplementation. To do this, we used genome editing to knock out ATP citrate lyase (ACLY), the enzyme responsible for converting citrate to oxaloacetate and acetyl-CoA in the cytoplasm and nucleus. In our cellular model, removing acetate from the culture medium resulted in the rapid catabolism of acetylated histones to restore the nucleocytoplasmic acetyl-CoA pool. Here we report the use of our new model in functional proteomics studies to characterize BRD-dependent interaction networks on the chromatin.

O-Acetyl-L-homoserine production enhanced by pathway strengthening and acetate supplementation in Corynebacterium glutamicum

BackgroundO-Acetyl-L-homoserine (OAH) is an important potential platform chemical. However, low levels of production of OAH are greatly limiting its industrial application. Furthermore, as a common and safe amino acid-producing strain, Corynebacterium glutamicum has not yet achieved efficient production of OAH.ResultsFirst, exogenous L-homoserine acetyltransferase was introduced into an L-homoserine-producing strain, resulting in the accumulation of 0.98 g/L of OAH. Second, by comparing different acetyl-CoA biosynthesis pathways and adding several feedstocks (acetate, citrate, and pantothenate), the OAH titer increased 2.3-fold to 3.2 g/L. Then, the OAH titer further increased by 62.5% when the expression of L-homoserine dehydrogenase and L-homoserine acetyltransferase was strengthened via strong promoters. Finally, the engineered strain produced 17.4 g/L of OAH in 96 h with acetate as the supplementary feedstock in a 5-L bioreactor.ConclusionsThis is the first report on the efficient production of OAH with C. glutamicum as the chassis, which would provide a good foundation for industrial production of OAH.