Intracellular Carbon Research Articles

Towards a high-rate operation of contact stabilization process: A microscopic view of carbon capture properties

Carbon capture performance is a key factor determining the chemical energy recovery potential of the high-rate contact stabilization (HiCS) process. However, the mechanisms of organic carbon capture are complex, involving surface adsorption, extracellular adsorption, and intracellular storage. A unique characteristic of the HiCS process is its low sludge residence time (SRT). Unfortunately, the influence of SRT on carbon capture has not been thoroughly studied, especially in terms of the underlying mechanisms. In this study, the microscopic changes in carbon capture performance during the transition from a conventional contact stabilized (CS) system to a high-rate mode of operation were demonstrated using intracellular carbon sources, extracellular polymeric substances (EPS), signaling molecules, and microbial community assays. The results showed that the extracellular carbon adsorption and intracellular carbon storage performance increased, and the microbial community structure changed significantly with converting the CS system to the high-rate operation mode. The enhancement of extracellular carbon adsorption performance mainly relied on the growth of EPS, which was accomplished by the strong growth of the relative abundance of the dominant bacterial group Cloacibacterium within the HiCS system, offsetting the negative effect produced by the decline of acyl-homoserine lactones. 98 mgCOD/gSS, 343 mgCOD/gSS, and 500 mgCOD/gSS of polyhydroxyalkanoates (PHAs) per sludge unit were obtained at SRT-24d, 8d, and 2d, respectively, suggesting that the HiCS system is more advantageous for rapid PHAs production.

Response to the CO2 concentrating mechanisms and transcriptional time series analysis of Ulva prolifera under inorganic carbon limitation

Ulva prolifera is a dominant species in green tides and has been affecting marine ecosystem for many years. Due to the low availability of CO2 in the environment, U. prolifera utilizes the CO2 concentrating mechanisms (CCMs) to increase intracellular inorganic carbon concentration. However, the transcriptional response mechanism and temporal changes of U. prolifera CCMs based on transcriptomics have not been thoroughly described. Therefore, we induced U. prolifera CCMs in a low CO2 environment to explore the dynamic regulation of CCMs expression under inadequate inorganic carbon supply. The results showed that inorganic carbon limitation increased the inorganic carbon affinity of U. prolifera, upregulating CCMs. The first 24 h of inorganic carbon environmental changes were the most active period for U. prolifera's expression regulation. U. prolifera gradually achieved a new steady state by regulating metabolic processes such as nucleic acids, energy, and ethylene-activated signaling pathways. In the carbon fixation system of U. prolifera, there are characteristics of both biophysical and biochemical CCMs. After 24 h of inorganic carbon limitation, the biophysical CCMs becomes more effective under conditions of inorganic carbon depletion. This study aids in exploring the CCMs of U. prolifera and their evolution in response to environmental changes.

Optimizing operation strategy to improve storage of intracellular carbon sources in anaerobic/oxic/anoxic system: Enhanced nitrogen removal by endogenous denitrification

Endogenous denitrification (ED) can make full use of the carbon sources and avoid replenishment of it. However, strengthening the storage of intracellular carbon sources is an important factor in improving ED efficiency. In this study, employed batch experiments using real domestic wastewater in the anaerobic/oxic (A/O) process. The anaerobic and oxic processes were run for 4 h under ambient conditions with the dissolved oxygen (DO) concentrations in the oxic stage controlled at 0.5, 1.0, 1.5, and 3.0 mg/L, respectively. The results showed that the content of poly-β-hydroxyalkanoates (PHA) reached its peak at 60 min (1.25 mmolC/L). And with DO concentrations of 1.5 mg/L, the contents of glycogen (Gly) were 27.74 mmolC/L. Subsequently, the AOA-SBR was established to investigate its effect on the long-term nitrogen removal performance of domestic wastewater by optimizing the anaerobic time and DO concentrations. The results showed that at an anaerobic time of 60 min and DO concentration of 1.5 mg/L, the storage of the intracellular carbon sources was highest and the total nitrogen (TN) removal efficiency increased to 82.12%. In addition, Candidatus Competibacter dominated gradually in the system as the strategy was optimized.

Intracellular carbon monoxide release unveils selective antibacterial effects

Intracellular carbon monoxide release unveils selective antibacterial effects

New insight into enhanced carbon recovery from anaerobic fermentation of waste activated sludge with cation exchange resin coupled with NaCl pretreatment

Carbon recovery from waste activated sludge has been attracting considerable attention. However, the migration and transformation patterns of carbon sources between the phases have rarely been reported. In this study, a novel strategy using cation exchange resin (CER) coupled with sodium chloride (NaCl) to enhance carbon recovery through anaerobic fermentation (AF) was proposed. The results demonstrated that CER coupled with NaCl destroyed OH and CO stretching in amide I while promoting the formation of β-sheet and random coil structures, leading to sludge disintegration. This significantly improved the kinetics of endogenous carbon release, resulting in the release of 1146.33 mg/L of carbon from the solid sludge into the liquid phase. Approximately 75.61 % of the initial carbon source was bio-transformed into short-chain fatty acids. Correspondingly, carbon recovery was significantly increased up to 852.23 mg C/L, 4.57 times that of the control. Mechanism exploration revealed that carbon source recovery was significantly elevated by the synergistic effect of CER and NaCl. CER effectively removed high-valence cations from extracellular polymeric substance (EPS), weakening its bridging and adsorption-electro neutralization capabilities, promoting protein deflocculation, and triggering EPS disruption to release extracellular carbon sources. NaCl disrupted the ionic strength and distribution inside and outside microbial cells, creating an osmotic pressure difference that resulted in cell plasmolysis and lysis, ultimately inducing the release of intracellular carbon sources. Economic and carbon emission reduction benefit analyses verified that the CER coupled with NaCl pretreatment is a cost-effective sludge treatment strategy. This study illustrates the carbon source migration and transformation pathways in the CER coupled with NaCl-assisted AF process, providing guidance for sustainable sludge management.

Interplay between photosynthetic electron flux and organic carbon sinks in sucrose-excreting Synechocystis sp. PCC 6803 revealed by omics approaches

BackgroundAdvancing the engineering of photosynthesis-based prokaryotic cell factories is important for sustainable chemical production and requires a deep understanding of the interplay between bioenergetic and metabolic pathways. Rearrangements in photosynthetic electron flow to increase the efficient use of the light energy for carbon fixation must be balanced with a strong carbon sink to avoid photoinhibition. In the cyanobacterium Synechocystis sp. PCC 6803, the flavodiiron protein Flv3 functions as an alternative electron acceptor of photosystem I and represents an interesting engineering target for reorganizing electron flow in attempts to enhance photosynthetic CO2 fixation and increase production yield.ResultsWe have shown that inactivation of Flv3 in engineered sucrose-excreting Synechocystis (S02:Δflv3) induces a transition from photoautotrophic sucrose production to mixotrophic growth sustained by sucrose re-uptake and the formation of intracellular carbon sinks such as glycogen and polyhydroxybutyrate. The growth of S02:Δflv3 exceeds that of the sucrose-producing strain (S02) and demonstrates unforeseen proteomic and metabolomic changes over the course of the nine-day cultivation. In the absence of Flv3, a down-regulation of proteins related to photosynthetic light reactions and CO2 assimilation occurred concomitantly with up-regulation of those related to glycolytic pathways, before any differences in sucrose production between S02 and S02:Δflv3 strains were observed. Over time, increased sucrose degradation in S02:Δflv3 led to the upregulation of respiratory pathway components, such as the plastoquinone reductase complexes NDH-11 and NDH-2 and the terminal respiratory oxidases Cyd and Cox, which transfer electrons to O2. While glycolytic metabolism is significantly up-regulated in S02:Δflv3 to provide energy for the cell, the accumulation of intracellular storage compounds and the increase in respiration serve as indirect sinks for photosynthetic electrons.ConclusionsOur results show that the presence of strong carbon sink in the engineered sucrose-producing Synechocystis S02 strain, operating under high light, high CO2 and salt stress, cannot compensate for the lack of Flv3 by directly balancing the light transducing source and carbon fixing sink reactions. Instead, the cells immediately sense the imbalance, leading to extensive reprogramming of cellular bioenergetic, metabolic and ion transport pathways that favor mixotrophic growth rather than enhancing photoautotrophic sucrose production.

Effect of Carbon Source on Endogenous Partial Denitrification Process: Characteristics of Intracellular Carbon Transformation and Nitrite Accumulation

This study focused on the start-up and operating characteristics of the endogenous partial denitrification (EPD) process with different carbon sources. Two sequencing batch reactors (SBRs) with sodium acetate (SBR1#) and glucose (SBR2#) as carbon sources were operated under anaerobic/oxic (A/O) and anaerobic/anoxic/oxic (A/A/O) modes successively for 240 d. The results showed that COD removal efficiency reached 85% and effluent COD concentrations were below 35 mg/L in both SBRs. The difference was that faster absorption and transformation of sodium acetate was achieved compared to glucose (COD removal rate (CRR) was 7.54 > 2.22 mgCOD/(L·min) in SBR1# compared to SBR2#). EPD could be started up with sodium acetate and glucose as carbon sources, respectively, and desirable high nitrite accumulations were both obtained at influent NO3−−N (NO3−-Ninf) increased from 20 to 40 mg/L with nitrate-to-nitrite transformation ratio (NTR) and specific NO3−-N deduction rate (rNa) of 88.4~90% and 2.41~2.38 mgN/(gVSS·h), respectively. However, at NO3−-N of 50~60 mg/L, both the NTR and rNa in SBR1# were higher compared to SBR2# (86.5% > 83.9% and 1.58 > 1.20 mgN/(gVSS·h), respectively). Hereafter, when NO3−-N was increased by 70~90 mg/L, lower NTR and rNa were observed in SBR1# than in SBR2# (72% and 78%, 1.16 and 1.32 mgN/(gVSS·h), respectively). Additionally, similar internal carbon transformations were observed to drive EPD for NO2−−N accumulation, especially for higher and faster carbon transformation with sodium acetate as carbon source compared to glucose. However, precise control of anoxic time as the peak point of nitrite (TNi,max) was still the key to achieve high NO2−−N accumulation.

Utilization of palm oil and palm oil effluent as a carbon source for the production of polyhydroxybutyrate co‐polyhydroxyhexanoate P(HB‐co‐HHx) a biopolymer in bacillus sp

AbstractBACKGROUNDBiodegradable polymers are gaining a reputation as demand is increasing globally. Amongst these Polyhydroxyalkanoates (PHA) are attracting market attention as a consequence of their properties similar to those of conventional plastics. PHA is a natural polyester that is stored as an intracellular carbon (C) energy reserve of bacteria. This biopolymer can be extracted and used in place of petrochemical plastic. Medium chain‐length (mcl) PHA, specifically poly hydroxybutyrate‐co‐hydroxyhexanoate P (HB‐co‐HHx) copolymer produced by Bacillus, has gained practical attention owing to its superior physicochemical properties compared to the commonly synthesized PHB homopolymer. Co‐polymer production is dependent on C sources provided in the growth medium. In the present study we have employed palm oil and palm oil effluent as C source.RESULTSIn the current study, it was observed that a significant amount of PHA and biomass was produced when the effluent was prepared at a concentration of 100% instead of water for preparing the growth medium. The production of biomass and PHA showed a range of 4.8–8and 1.5–4 g L−1, respectively. This means that the PHA yield obtained using the effluent was 50–80% of the control, whereas the control yielded only 40–50%.CONCLUSIONPalm industry waste can be used as a substrate in bioprocessing industries. Proton nuclear magnetic resonance analysis of PHA obtained herein indicates that it consists of the mcl polymer P (HB‐co‐HHx). © 2024 The Author(s). Journal of Chemical Technology and Biotechnology published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry (SCI).

Exploring the carbon skeleton cycling regulation mechanism of single-wavelength light quality promoting the accumulation of polysaccharides in Spirulina platensis FACHB-439: Based on transcriptomics

Carbon dioxide can be converted into carbohydrates by Spirulina to promote carbon neutrality and produce high added value bio-functional compounds (e.g. polysaccharides). This study explored the effects of different single-wavelength light qualities on Spirulina polysaccharides and the differences in gene expression. The experiment results indicated that red light could significantly increase the content of polysaccharides in Spirulina (116.88 mg/g), followed by blue light (99.83 mg/g), green light (95.23 mg/g), yellow light (50.50 mg/g), while the white light control group was 69.59 mg/g. Transcription analysis results revealed that red light adjusted the intracellular carbon skeleton cycling between glycolysis/gluconeogenesis pathway, pyruvate and amino acid metabolism, and protein synthesis, ultimately promoting the accumulation of carbon sources towards polysaccharide synthesis. These findings offer theoretical basis and data support for the promising carbon fixation and reutilization based on blue-green algae.

Metabolomic reveals the responses of sludge properties and microbial communities to high nitrite stress in denitrifying phosphorus removal systems

Nitrite, as an electron acceptor, plays a good role in denitrifying phosphorus removal (DPR); however, high nitrite concentration has adverse affects on sludge performance. We investigated the precise mechanisms of responses of sludge to high nitrite stress, including surface characteristics, intracellular and extracellular components, microbial and metabolic responses. When the nitrite stress reached 90 mg/L, the sludge settling performance was improved, but the activated sludge was aging. FTIR and XPS analysis revealed a significant increase in the hydrophobicity of the sludge, resulting in improve settling performance. However, the intracellular carbon sources synthesis was inhibited. In addition, the components in the tightly bound extracellular polymeric substances (TB-EPS) of sludge were significantly reduced and indicated the disturb of metabolism. Notably, Exiguobacterium emerged as a new genus when face high nitrite stress that could maintaining survival in hostile environments. Moreover, metabolomic analysis demonstrated strong biological response to nitrite stress further supported above results that include the inhibited of carbohydrate and amino acid metabolism. More importantly, some lipids (PS, PA, LysoPA, LysoPC and LysoPE) were significantly upregulated that related enhanced membrane lipid remodeling. The comprehensive analyses provide novel insights into the high nitrite stress responses mechanisms in activated sludge systems.

Size-dependent dynamics of the internal carbon pool drive isotopic vital effects in calcifying phytoplankton

Isotopic offsets in biogenic calcite from equilibrium values can provide unique insights into the physiology and mechanisms of carbon regulation in calcifying phytoplankton. This study examines the impact of varying CO2 (controlled via pH) on five coccolithophore species chosen for varied cell sizes, physiology, and calcification. The study investigates isotopic offsets in coccolith calcite and organic matter, in relation to carbon demand and supply (µ/CO2). Species-specific CO2 and/or pH optima for growth (µopt) were derived from variations in growth rates with varying CO2 concentrations. Growth rates for all species declined with rising CO2 (decreasing pH) due to H+-driven inhibition. C. braarudii and C. leptoporus exhibited µopt at high CO2 concentrations (suggesting high carbon-demand) and limited growth under low CO2 (high pH) suggesting carbon limitation. Under low CO2 supply, when growth rates were CO2-limited, both species exhibited coincident isotopic depletion in calcite and organic matter as a consequence of CO2 diffusion into the cell that experienced no equilibration as a result of a highly depleted internal carbon pool. In these two species, isotopic values in calcite remained unaffected by growth rates and CO2 concentration (µ/CO2) when CO2 was sufficient for optimal growth. G. huxleyi and G. oceanica displayed optima for growth (µopt) at low CO2 concentrations and showed no growth limitation under low CO2 (indicating low carbon-demand). Both species experienced depleted (negative) vital effects caused by an excess of CO2 diffusioninto the small internal carbon pool of the cell when diffusive carbon supply outpaced low demand (low µ/CO2). Enriched (positive) vital effects were observed under low carbon supply and high demand, likely due to increased HCO3– uptake and diffusive CO2 loss from the intracellular carbon pooldue to a lower intracellular pH than the seawater pH. C. carterae exhibited a µopt at intermediate CO2 concentrations and isotopically equilibrated intracellular carbon pool such that δ13C values in calcite and organic matter suggested a shared carbon pool. This study illustrates that pH and CO2 driven vital effects and fractionation into organic matter indicate the residence time for carbon in the intracellular carbon pool, where the size of the pool is proportional to cell size. Due to the increased buffering afforded by a larger pool, C. leptoporus and C. carterae may have elevated intracellular pH which minimises CO2 leakage, whereas vital effects in G. huxleyi and G. oceanica are caused by CO2 diffusion in or out of their small internal carbon pool with limited buffering capacity owing to its small size.

De Novo Synthesis of Chrysin in Saccharomyces cerevisiae.

Chrysin, a flavonoid, has been found to have been widely used in the health food field. But at present, chrysin production is hindered by the low availability of precursors and the lack of catalytic enzymes with high activity. Therefore, ZmPAL was initially screened to synthesize trans-cinnamic acid with high catalytic activity and specificity. To enhance the supply of precursors, the shikimic acid and chorismic acid pathway genes were overexpressed. Besides, the expression of the intracellular and mitochondrial carbon metabolism genes CIT, MAC1/3, CTP1, YHM2, RtME, and MDH was enhanced to increase the intracellular acetyl-CoA content. Chrysin was synthesized through a novel gene combination of ScCPR-EbFNSI-1 and PcFNSI. Finally, de novo synthesis of chrysin was achieved, reaching 41.9 mg/L, which is the highest reported concentration to date. In summary, we identified efficient enzymes for chrysin production and increased it by regulating acetyl-CoA metabolism in mitochondria and the cytoplasm, laying a foundation for future large-scale production.

Allosteric regulation of nitrate transporter NRT via the signaling protein PII

Coordinated carbon and nitrogen metabolism is crucial for bacteria living in the fluctuating environments. Intracellular carbon and nitrogen homeostasis is maintained by a sophisticated network, in which the widespread signaling protein PII acts as a major regulatory hub. In cyanobacteria, PII was proposed to regulate the nitrate uptake by an ABC (ATP-binding cassette)-type nitrate transporter NrtABCD, in which the nucleotide-binding domain of NrtC is fused with a C-terminal regulatory domain (CRD). Here, we solved three cryoelectron microscopy structures of NrtBCD, bound to nitrate, ATP, and PII, respectively. Structural and biochemical analyses enable us to identify the key residues that form a hydrophobic and a hydrophilic cavity along the substrate translocation channel. The core structure of PII, but not the canonical T-loop, binds to NrtC and stabilizes the CRD, making it visible in the complex structure, narrows the substrate translocation channel in NrtB, and ultimately locks NrtBCD at an inhibited inward-facing conformation. Based on these results and previous reports, we propose a putative transport cycle driven by NrtABCD, which is allosterically inhibited by PII in response to the cellular level of 2-oxoglutarate. Our findings provide a distinct regulatory mechanism of ABC transporter via asymmetrically binding to a signaling protein.

Calcium isotope fractionation by intracellular amorphous calcium carbonate (ACC) forming cyanobacteria.

The formation of intracellular amorphous calcium carbonate (ACC) by various cyanobacteria is a widespread biomineralization process, yet its mechanism and importance in past and modern environments remain to be fully comprehended. This study explores whether calcium (Ca) isotope fractionation, linked to ACC-forming cyanobacteria, can serve as a reliable tracer for detecting these microorganisms in modern and ancient settings. Accordingly, we measured stable Ca isotope fractionation during Ca uptake by the intracellular ACC-forming cyanobacterium Cyanothece sp. PCC 7425. Our results show that Cyanothece sp. PCC 7425 cells are enriched in lighter Ca isotopes relative to the solution. This finding is consistent with the kinetic isotope effects observed in the Ca isotope fractionation during biogenic carbonate formation by marine calcifying organisms. The Ca isotope composition of Cyanothece sp. PCC 7425 was accurately modeled using a Rayleigh fractionation model, resulting in a Ca isotope fractionation factor (Δ44Ca) equal to -0.72 ± 0.05‰. Numerical modeling suggests that Ca uptake by these cyanobacteria is primarily unidirectional, with minimal back reaction observed over the duration of the experiment. Finally, we compared our Δ44Ca values with those of other biotic and abiotic carbonates, revealing similarities with organisms that form biogenic calcite. These similarities raise questions about the effectiveness of using the Ca isotope fractionation factor as a univocal tracer of ACC-forming cyanobacteria in the environment. We propose that the use of Δ44Ca in combination with other proposed tracers of ACC-forming cyanobacteria such as Ba and Sr isotope fractionation factors and/or elevated Ba/Ca and Sr/Ca ratios may provide a more reliable approach.

Metabolic shifts in glycogen-accumulating organisms: Exploring carbon and phosphorus dynamics under two distinct phosphorus limitation modes

Glycogen-accumulating organisms (GAOs) are recognized as competitors of phosphorus-accumulating organisms (PAOs) in enhanced biological phosphorus removal (EBPR) systems. However, different types of GAOs exhibit unique metabolic traits in different environments. To characterize the metabolic activity of GAOs, two distinct modes were constructed. Long-term operation with the mode of anaerobic post-drainage demonstrated the successful establishment of a rapid and enduring GAOs system dominated by Candidatus Contendobacter community. Acetate was completely consumed within 60 min of the anaerobic phase. Concurrently, glycogen levels decreased by 0.062 g Glycogen/g SS, while intracellular carbon storage of poly-hydroxybutyrate (PHB) and poly-hydroxyvalerate (PHV) increased by 0.027 g PHB/g SS and 0.007 g PHV/g SS, respectively. Various concentrations of phosphorus loads illustrated the resistance of the system to high phosphorus loads, while maintaining glycogen accumulating metabolism (GAM). Under complete phosphorus limitation, no correlation between glycogen and acetate was observed during the initial 140 d. The addition of 1 mg/L phosphate to the influent preserved the typical GAM characteristics. Moreover, the pathway involved in glycogen conversion to trehalose may be pivotal in enhancing anaerobic glycogen degradation. The complete tricarboxylic acid cycle ensured efficient energy production, thereby sustaining the foundational metabolism. Notably, sucC and mdh were critical regulators of energy flux. This study provides a distinctive metabolic profile of a robust and resilient GAOs model with Ca. Contendobacter.

AlphaFold2-guided description of CoBaHMA, a novel family of bacterial domains within the heavy-metal-associated superfamily.

Three-dimensional (3D) structure information, now available at the proteome scale, may facilitate the detection of remote evolutionary relationships in protein superfamilies. Here, we illustrate this with the identification of a novel family of protein domains related to the ferredoxin-like superfold, by combining (i) transitive sequence similarity searches, (ii) clustering approaches, and (iii) the use of AlphaFold2 3D structure models. Domains of this family were initially identified in relation with the intracellular biomineralization of calcium carbonates by Cyanobacteria. They are part of the large heavy-metal-associated (HMA) superfamily, departing from the latter by specific sequence and structural features. In particular, most of them share conserved basic amino acids(hence their name CoBaHMA for Conserved Basic residues HMA), forming a positively charged surface, which is likely to interact with anionic partners. CoBaHMA domains are found in diverse modular organizations in bacteria, existing in the form of monodomain proteins or as part of larger proteins, some of which are membrane proteins involved in transport or lipid metabolism. This suggests that the CoBaHMA domains may exert a regulatory function, involving interactions with anionic lipids. This hypothesis might have a particular resonance in the context of the compartmentalization observed for cyanobacterial intracellular calcium carbonates.

Fatty Acid Metabolism in Peroxisomes and Related Disorders.

One of the functions of peroxisomes is the oxidation of fatty acids (FAs). The importance of this function in our lives is evidenced by the presence of peroxisomal disorders caused by the genetic deletion of proteins involved in these processes. Unlike mitochondrial oxidation, peroxisomal oxidation is not directly linked to ATP production. What is the role of FA oxidation in peroxisomes? Recent studies have revealed that peroxisomes supply the building blocks for lipid synthesis in the endoplasmic reticulum and facilitate intracellular carbon recycling for membrane quality control. Accumulation of very long-chain fatty acids (VLCFAs), which are peroxisomal substrates, is a diagnostic marker in many types of peroxisomal disorders. However, the relationship between VLCFA accumulation and various symptoms of these disorders remains unclear. Recently, we developed a method for solubilizing VLCFAs in aqueous media and found that VLCFA toxicity could be mitigated by oleic acid replenishment. In this chapter, we present the physiological role of peroxisomal FA oxidation and the knowledge obtained from VLCFA-accumulating peroxisome-deficient cells.

Characterization the response of Chlamydomonas reinhardtii serine/threonine protein kinase mutant to blue light

In order to investigate the molecular mechanism of silk/threonine protein kinase (STK)-mediated blue light response in the algal Chlamydomonas reinhardtii, phenotype identification and transcriptome analysis were conducted for C. reinhardtii STK mutant strain crstk11 (with an AphvIII box reverse insertion in stk11 gene coding region) under blue light stress. Phenotypic examination showed that under normal light (white light), there was a slight difference in growth and pigment contents between the wild-type strain CC5325 and the mutant strain crstk11. Blue light inhibited the growth and chlorophyll synthesis in crstk11 cells, but significantly promoted the accumulation of carotenoids in crstk11. Transcriptome analysis showed that 860 differential expression genes (DEG) (559 up-regulated and 301 down-regulated) were detected in mutant (STK4) vs. wild type (WT4) upon treatment under high intensity blue light for 4 days. After being treated under high intensity blue light for 8 days, a total of 1 088 DEGs (468 upregulated and 620 downregulated) were obtained in STK8 vs. WT8. KEGG enrichment analysis revealed that compared to CC5325, the crstk11 blue light responsive genes were mainly involved in catalytic activity of intracellular photosynthesis, carbon metabolism, and pigment synthesis. Among them, upregulated genes included psaA, psaB, and psaC, psbA, psbB, psbC, psbD, psbH, and L, petA, petB, and petD, as well as genes encoding ATP synthase α, β and c subunits. Downregulated genes included petF and petJ. The present study uncovered that the protein kinase CrSTK11 of C. reinhardtii may participate in the blue light response of algal cells by mediating photosynthesis as well as pigment and carbon metabolism, providing new knowledge for in-depth analysis of the mechanism of light stress resistance in the algae.

Metabolic flux reprogramming in Mycobacterium tuberculosis-infected human macrophages.

Metabolic fluxes are at the heart of metabolism and growth in any living system. During tuberculosis (TB) infection, the pathogenic Mycobacterium tuberculosis (Mtb) adapts its nutritional behaviour and metabolic fluxes to survive in human macrophages and cause infection. The infected host cells also undergo metabolic changes. However, our knowledge of the infected host metabolism and identification of the reprogrammed metabolic flux nodes remains limited. In this study, we applied systems-based 13C-metabolic flux analysis (MFA) to measure intracellular carbon metabolic fluxes in Mtb-infected human THP-1 macrophages. We provide a flux map for infected macrophages that quantified significantly increased fluxes through glycolytic fluxes towards pyruvate synthesis and reduced pentose phosphate pathway fluxes when compared to uninfected macrophages. The tri carboxylic acid (TCA) cycle fluxes were relatively low, and amino acid fluxes were reprogrammed upon Mtb infection. The knowledge of host metabolic flux profiles derived from our work expands on how the host cell adapts its carbon metabolism in response to Mtb infection and highlights important nodes that may provide targets for developing new therapeutics to improve TB treatment.

Targeted synergy of cation exchange-driven extracellular carbon release and lysozyme-catalyzed intracellular carbon release for maximizing carbon source recovery from waste activated sludge

Short-chain fatty acids as superior carbon recovery from waste activated sludge have received rising attentions. This study quantified contributions of cation exchange (CE) and lysozyme on the targeted carbon source migration. The innovative targeted pattern for extracellular and intracellular carbon source release was proposed. The 1-day synergetic treatment at optimal CE resin dosage of 1.75 g/g SS and lysozyme dosage of 10 % drove considerable carbon source release and particulate carbon source solubilization, which occupied 35.31 % of total sludge solid carbon sources. The CE dominantly contributed to extracellular polymeric substances disruption and extracellular carbon source release, with the percentage of 39.47–48.13 %. In this case, The Lyso was the strongest contributor to microbial cell lysis and intracellular carbon source release, with the proportions of 49.53–75.76 %. Subsequently, approximately 56.54–58.49 % of the originally dissolved carbon sources (mostly proteins and carbohydrates) were bio-degraded into micromolecular carbon sources with high bioavailability in a short-term anaerobic fermentation (2 days). Correspondingly, satisfactory carbon recovery rate of 23.06 % was obtained, which was 7.30 times higher than the control. The synergy of CE and lysozyme greatly improved kinetic speeds and performances of endogenous carbon source migration and recovery. Such synergetic treatment of CE and lysozyme not only provided novel insights into carbon fate in anaerobic fermentation, but also achieved remarkable carbon source release and recovery benefits with economic incomes of 190.8–283.4 CNY/ton sludge.