synechococcus elongatus pcc 7942

This strain was constructed to more fully allow sucrose into the cell through the CscB transporter and to be hydrolyzed to glucose and fructose by the endogenous sucrose invertase (putatively encoded by SYNPCC7942_0397). BG-11 medium only was used as a blank, and excitation and emission wavelengths were set to 485 and 528 nm, respectively. Taxonomy - Synechococcus elongatus (strain PCC 7942 / FACHB-805) (Anacystis nidulans R2) shadow: false, Of them, KaiA, KaiB and KaiC comprise the central oscillator components, which are … Summary: addSiteSpecificMenuItems(); The strain into which the synthetic metabolic pathway was introduced produced 3.48 mM (0.265 g/L) … The strain containing glcP showed increased growth (0.730 day−1) during the first light phase, but the growth stopped after that, which is consistent with the previous results (Fig. Thus, using HPLC analysis, there was a high level of difficulty in detecting such a small change in the glucose concentration over a 96-h period while taking into account evaporation of water. Trophic conversion of an obligate photoautotrophic organism through metabolic engineering, Obligate phototrophy in cyanobacteria: more than a lack of sugar transport, Identification of a metabolic network structure representative of Arthrospira (Spirulina) platensis metabolism, The tricarboxylic acid cycle in cyanobacteria, Rerouting carbon flux to enhance photosynthetic productivity, Engineering cyanobacteria to synthesize and export hydrophilic products, Chemoheterotrophic growth of the cyanobacterium, Regulation of fructose transport and its effect on fructose toxicity in, An engineered microbial platform for direct biofuel production from brown macroalgae, Direct photosynthetic recycling of carbon dioxide to isobutyraldehyde, Fatty acid production in genetically modified cyanobacteria, Bioactive natural products from marine cyanobacteria for drug discovery, Generic assignments, strain histories and properties of pure cultures of cyanobacteria, Non-fermentative pathways for synthesis of branched-chain higher alcohols as biofuels, LdpA: a component of the circadian clock senses redox state of the cell, pBBR1MCS: a broad-host-range cloning vector, Genetic engineering of the cyanobacterial chromosome, From bacterial glycogen to starch: understanding the biogenesis of the plant starch granule, Bacterial glycogen synthesis and its regulation, Modes of cyanobacterial carbon metabolism, Carbohydrate metabolism in mutants of the cyanobacterium, Molecular and genetical analysis of the fructose-glucose transport system in the cyanobacterium, Sequence and structure of a human glucose transporter, The utilization of organic substrates by cyanobacteria, Microbial production of fatty-acid-derived fuels and chemicals from plant biomass, Alternative biofuel production in non-natural hosts, Light-responsive gene expression in cyanobacteria, Quinone sensing by the circadian input kinase of the cyanobacterial circadian clock, Thioredoxin-linked processes in cyanobacteria are as numerous as in chloroplasts, but targets are different, CikA, a bacteriophytochrome that resets the cyanobacterial circadian clock, Light-activated heterotrophic growth of the cyanobacterium, Submission, Review, & Publication Processes, Engineering Synechococcus elongatus PCC 7942 for Continuous Growth under Diurnal Conditions. 3C). Glucose and xylose consumption assays.Glucose and xylose concentrations in the culture medium were measured using a high-performance liquid chromatograph (Shimadzu) equipped with an Aminex HPX-87 column (Bio-Rad) and a refractive index detector. The location of integration into the chromosome (neutral site, NS1) has been developed as a cloning locus (Clerico. Optimum growth resulted from induction using 0.1 mM IPTG, while induction with a 1 mM concentration led to a slightly lower cell growth rate (Fig. The complete segregation of ΔglgC was confirmed by PCR using the primers IM171 and GR015 (see Fig. Direct conversion of carbon dioxide (CO 2) to value-added chemicals by engineering of cyanobacteria has received attention as a sustainable strategy in food and chemical industries. Additionally, by constraining photon uptake fluxes, we … PCC 7942 relies on homologous recombination between the cell’s chromosome and exogenous DNA that is not autonomously replicating and containing sequences homologous to the chromosome. (C) Growth curves of the cscB-cscK strain (red) and the wild type (blue). Synechococcus, */ 4B). The PCR product was digested with SalI and NheI and ligated with pGR01 cut with the same enzyme, creating pAL82. The y axis indicates GFP fluorescence intensity divided by OD730. Error bars represent standard deviations (in triplicate). The well-known photoheterotrophic cyanobacterium Synechocystis sp. Some proteins have been identified that are involved in circadian signal transduction in S. elongatus. Quantitative real-time PCR (qPCR) technology is the most commonly used method for studying the expression of specific genes, in which the relative expression level of target … Zaslavskaia et al. It has also been widely used as a bioreactor to produce biofuels and other metabolic products. Journal of Microbiology & Biology Education, Microbiology and Molecular Biology Reviews. The Thus, we hypothesize that efficient uptake of sugars is the major missing factor inhibiting heterotrophy in obligate photoautotrophic cyanobacteria. Synechococcus elongatus PCC 7942 is a model freshwater cyanobacterium. It was this isolate, previously known as Anacystis nidulans R2, that was the first cyanobacterium demonstrated to be reliably transformable by exogenously added DNA (32). Correct recombinants were confirmed by PCR and DNA sequencing to verify integration of targeting genes into the chromosome at NSI (see Fig. These results suggest that carbon fixation is not the rate-limiting step for the growth of the galP strain in the presence of 5 g/liter glucose, while carbon fixation is growth limiting for the wild type under the same conditions. In S. elongatus, light regulation takes place through two major routes: transcriptional and posttranslational. As a result, the overall productivity of a photoautotrophic system is low. PCC 7002 (Ss7002), characterized by the production of trienoic alpha-linolenic acid (ALA, C18:3 Δ 9,12,15) by Δ9 … PCC 6803 (28), GalP from Escherichia coli (29), and Glut1 from human erythrocytes (30). updateBannerClass(); Red arrows indicate steps catalyzed by heterologous enzymes. Enter multiple addresses on separate lines or separate them with commas. The laser percentage was set to 11.5%. // User sites can define this fn in userWebsiteCustomization.js in The standardized fluorescent intensity (relative fluorescence units [RFU]/OD730) indicated that 1 mM IPTG and 0.1 mM IPTG in the medium caused similar levels of expression of GalP-GFP, and expression from Ptrc was saturated with 0.1 mM IPTG for this construct. onMenuBarReady.fire([]); Sucrose is another important feedstock that is produced on the economic scale needed for common human consumption, as well as being utilized for biologically derived fuels and chemicals. While the universal causes of obligate photoautotrophy may be diverse, installing sugar transporters provides new insight into the mode of obligate photoautotrophy for cyanobacteria. PCC 6803 genomic DNA (ATCC). Assays were examined for contamination by plating 50 μl of the culture from the final time point of every sample on Luria Bertani (LB) plates. 2E). 4C). 1B). Xylose isomerase and xylulokinase are responsible for the first two steps of xylose degradation (Fig. The Synechococcus elongatus PCC 7942 genome contains 2719 genes, which we have assigned computationally to 1964 transcription units. The galP-ΔglgC strain failed to show any growth in the presence of glucose beyond the first light cycle (Fig. The activities of all photosynthetic organisms are energy intensive, interconnected with central metabolism, and are strictly regulated according to light intensity. Next, various concentrations of IPTG were added to the cultures of the galP-gfp strain, and the growth was measured under continuous-light conditions (Fig. Synechococcus elongatus PCC 7942 is a model freshwater cyanobacterium. Installation of the xylose degradation pathway. After the initial 12 h, the growth rates of the two strains were comparable in the light. The correct deletion of glgC was confirmed by PCR with the primers GR050 and IM581 and IM573 and GR015 (see Fig. Ron Caspi1, In a few of the samples, slight contamination was detected (<20 colonies out of at least 104 cyanobacterial cells). includes: Synechococcus elongatus PCC 7942. // Add dynamically all the menubar commands and menus here if need be. The metabolic changes coincided with a general reprogramming of gene expression, which … Database lists are saved as SmartTables that you can edit from your, ©2019 SRI International, 333 Ravenswood Avenue, Menlo Park, CA 94025-3493. Terrabacteria group, The baseline growth rate of wild-type S. elongatus based on the OD730 was 0.161 day−1 during the light cycle (48 h to 60 h) with no growth during the dark cycle (Fig. In this work, we developed a strategy to allow growth of an obligate photoautotroph on diverse feedstocks. This is slightly faster than the fastest growth shown here for the galP strain under photomixotrophic conditions, with a growth rate of about 0.99 (day−1). However, more recently, it has been shown that two alternative enzymes, 2-oxoglutarate decarboxylase and succinic semialdehyde dehydrogenase, convert 2-oxoglutarate to succinate and functionally complete the TCA cycle in cyanobacteria (6). We can assume these variations are related to the ability of these membrane proteins in S. elongatus to be adequately expressed, folded, and localized, as well as differences in enzymatic activity. strain PCC 6803, has been expressed from a plasmid-based expression system in Synechococcus elongatus strain PCC 7942 but could not be maintained, while genomic integration led to glucose sensitivity (4). Author information: (1)Department of Biotechnology, Graduate School of Engineering, Nagoya University, Furo-cho, Chikusa-ku, Nagoya, Aichi, 464-8603, Japan. All preculture was grown without the addition of sodium bicarbonate or any saccharide. The effects of bicarbonate on heterotrophic growth were characterized (Fig. 2C). Interestingly, the maximum growth of the galP strain (0.992 ± 0.014) was faster than that of the xylEAB strain (0.572 ± 0.025) under the light condition in the presence of their respective sugars owing to rapid growth in the galP strain in the first 12 h (Fig. Supplemental material for this article may be found at http://dx.doi.org/10.1128/AEM.03326-12. To amplify the growth difference between strains that could and could not grow on extracellular glucose, conditions were set so as to limit the amount of CO2 (no bubbling) and light intensity (25 μE/m2/s). hidedelay: 700 YAHOO.util.Event.onAvailable("body", function () { Synechococcus elongatus strain PCC 7942 strictly depends upon the generation of photosynthetically derived energy for growth and is incapable of biomass increase in the absence of light energy. The only engineered strain to show a consistent increase in the growth rate in the presence of glucose contained galP (denoted as the galP strain) (Fig. Here, we investigated the therapeutic efficacy of this cyanobacterium in a mouse model of acute burn injury and whether the secretion of extracellular vesicles (EVs), important mediators of cell paracrine activity, is a key mechanism of the cyanobacterium … 2A and B). Probing the limitations of S. elongatus regarding the consumption of each of these common feedstocks gives a greater understanding of the more complex and general obligate photoautotrophic behavior, as well as aiding in the development of industrially relevant and versatile chemical-producing photosynthetic organisms. engineered a eukaryotic alga, Phaeodactylum tricornutum, to thrive on exogenous glucose in the absence of light by introducing a gene encoding a glucose transporter from human erythrocytes (Glut1 or Hup1) (3). In general, heterotrophic cyanobacteria utilize relatively few substrates for the buildup of biomass (18). Model cyanobacteria such as Synechococcus elongatus PCC 7942 and UTEX 2973, and Synechocystis sp. Other cultured cyanobacterial species are obligate photoautotrophs and as such lack the ability to consume fixed carbon compounds for increased biomass or energy. Research is investigating efficient harvesting of glucose from cellulose as a feedstock for biofuel production. White and shaded areas indicate light and dark cycles. 2A). Synechococcaceae, cscB and cscK were amplified using the primers JM55 and JM56, digested with EcoRI and BamHI, and ligated to similarly digested pAM2991 to create pAL289. However, the xylose transporter not only failed to improve the growth of the strain in the presence of xylose under diurnal conditions, but this mutant had a significantly reduced growth rate (0.154 ± 0.015) compared to that of the wild type (0.350 ± 0.017) (Fig. S. elongatus UTEX 2973 contains a SNP to the gene encoding ATP synthase F1 subunit α, comparable to the corresponding gene in Synechococcus PCC 7942. The growth rates of the galP strain under light and dark conditions in the presence of glucose were 0.540 day−1 and 0.199 day−1, respectively. Bacteria , Synechococcus elongatus PCC 7942 is a naturally occurring photoautotrophic cyanobacterium that can be easily obtained and large-scale expanded. GFP assay on plate reader.All green fluorescent protein (GFP) assays were done using a Microtek Synergy H1 plate reader (BioTek). Synechococcus elongatus PCC 7942 . Spectinomycin was purchased from MP Biomedicals (Santa Ana, CA). Glycogen is built up within the cell throughout the light phase of metabolism and then used as an energy source to maintain essential chemical processes throughout the dark phase (26). A 488-nm laser was used for excitation of all mutants. Hence, engineering the … Synechococcus elongatus strain PCC 7942 strictly depends upon the generation of photosynthetically derived energy for growth and is incapable of biomass increase in the absence of light energy. White and shaded areas indicate the light and dark cycles, respectively. However, the expression of xylE may greatly increase the amount of xylose accumulated within the cell owing to its slow metabolism, causing a metabolic imbalance. Cyanobacteria/Melainabacteria group, Most characterized cyanobacterial species are obligate photoautotrophs under examined conditions. on the screen. S1 in the supplemental material). (A) Schematic representation of integration of the the sucrose degradation pathway into the S. elongatus chromosome. For all reported results where contamination was detected, green cells were greater than 99% of the culture (n ≥ 500). Two principal hypotheses have been proposed for obligate photoautotrophy among microorganisms. /* S1C). Among them are proteins involved in the biogenesis and functioning of the cyanobacteria cell wall (Synpcc7942_1362, Synpcc7942_0351, Synpcc7942_0732) and protein participating in stress response at DNA restriction-modification level (Synpcc7942_0726). Glucose is a common energy storage molecule in S. elongatus in the form of glycogen (24–27). PPP, pentose phosphate pathway. Cyanobacteria are increasingly being studied for the production of valuable natural products, as well as biofuels and chemicals (35). NSI, neutral site 1. Since xylose slightly improved the growth of the wild type (Fig. Counting chambers were chosen randomly, and green cells versus colorless cells were tallied. Fluorescence intensity of the cultures of the galP-gfp strain was measured throughout the growth assay (Fig. … These transporters include GlcP from Synechocystis sp. Confocal microscopy.All confocal microscope images were taken using the Olympus_America FV1000 system located within the UCD NEAT Spectral Imaging Facility. glcP was amplified using the primers MC170 and MC171, digested with BamHI and EcoRI, and ligated to similarly digested pAM 2991 to create pAL46. NOTE: We request your email address only to inform the recipient that it was you who recommended this article, and that it is not junk mail. Thus, the difficulty of predicting heterologous enzymatic activity, especially for membrane proteins, remains high. {autosubmenudisplay: true, xylAB was amplified using the primers JM07 and JM08 and digested with AvrII and BamHI, followed by ligation to similarly digested pAL65 to create pAL70. // Render the MenuBar instance /* This code needs to be here to avoid the menu and other Yahoo widgets to flicker All primers used are listed in Table 2. Synechococcus elongatus PCC 7942 genome contains 2719 genes, which we have assigned computationally to 1964 transcription units. Throughout Results/Discussion, growth rate values reported from the light and dark cycles are derived from the 48-to-60-h and 60-to-72-h periods, respectively (growth rates for all periods, 0 to 96 h, are listed in Table 4). Both strains are closely related to each other. With the addition of bicarbonate in this experiment, the pH of the culture may increase as the bicarbonate is consumed. 1D). 1B and C). (C) Growth curve of the Glut1 (purple) and glcP (green) strains and the wild type. Moreover, cyanobacterial chemical production has gained increased attention. Info; Home; Download; Status; HELP! Herein, Synechococcus elongatus PCC 7942, a model cyanobacterium, was engineered to produce α-farnesene from CO 2. (B) Growth curve of the galP strain (red) and wild type (blue) with and without 5 g/liter glucose. These results indicate that GalP from E. coli is successfully localized to the membrane of S. elongatus and may allow efficient transport of glucose into the cell. Another possibility is exploring a light pulsing stratagem similar to the light-activated heterotrophic growth seen in Synechocystis sp. (A) Schematic representation of integration of the xylose degradation pathway into the S. elongatus chromosome. The galP, xylE, xylA, and xylB genes were amplified from Escherichia coli BW25113 genomic DNA. The strain containing galP-gfp showed a fluorescent signal only in the cellular membrane (Fig. The product was digested with XhoI and MluI and ligated with similarly digested pSA69 (19), creating pGR01. Obligate phototrophs' core metabolism is very similar to that of heterotrophic counterparts exhibiting diverse trophic behavior. 1B and Table 4). NCBI-Taxonomy:1140, Genetic Code Number: Several investigations to examine obligate photoautotrophy in cyanobacteria with recombinant sugar transporter strategies have been attempted, yet a conclusive determination of the causes of photoautotrophy has proven to be elusive. Synechococcus elongatus PCC 7942 (PCC 7942) is a cyanobacterium that is known to synthesize zeaxanthin as one of the predominant cellular carotenoids and does not possess pathway genes for lutein production. In future work, we will seek to engineer the strains from this work to allow growth under continuously dark conditions. All y axes denote OD730,though the scales differ for visibility. cscB and cscK were amplified from genomic DNA of E. coli ATCC 700927 (ATCC). It is an obligate photoautotroph, with a genome of approximately 2.7 Mb, and two endogenous plasmids, pANL and pANS. Sugar transporters, as well as other peripheral genes, were integrated for the increased growth rate of a model photoautotrophic cyanobacterium under both light and dark conditions. S1B). The wild type grew at a higher rate (0.204 day−1) when in the presence of glucose and light, yet exhibited no growth in the dark cycle (Fig. The transformation of . This study was supported by a National Science Foundation grant (1132442). The galP strain showed similar growth with or without various concentrations of bicarbonate- and glucose-containing medium in continuous light, while a higher concentration of bicarbonate slightly enhanced the growth of the wild type (Fig. It was found that the cscK-cscB strain had increased growth rates, 0.376 day−1 (light) and 0.128 day−1 (dark), in the presences of 5 g/liter sucrose (Fig. All samples are grown in BG-11 medium with 5 g/liter glucose in continuous light. These strains were assayed to determine any change in growth behavior. Synechococcus elongatus PCC 7942, Unification Links: Here we describe a methodology for constraining genome-scale models of photoautotrophy in the cyanobacteria Synechococcus elongatus PCC 7942. These results demonstrate that S. elongatus can metabolize glucose for growth once it is transported into the cells. Synechococcus elongatus PCC 7942 version 24.5. 4C). (B) Confocal microscope images of the gfp strain (left) and the galP-gfp strain (right). While the particular genes necessary for each specific feedstock may differ, these results suggest that the cause of autotrophy in this organism lies within the peripheral metabolic processes. 2B, left), and the wild type did not show a fluorescent signal under these conditions (Fig. We have determined one underlying cause of natural photoautotrophy in this bacterium by engineering the organism to allow diurnal growth. Recently, fast-growing cyanobacterial strains Synechococcus elongatus UTEX 2973 and S. elongatus PCC 11801 (henceforth UTEX 2973 and PCC 11801, respectively) have been reported 19, 20. YAHOO.util.Event.onContentReady("pathwayToolsMenubar", function () { (B) Synthetic sucrose degradation pathway in S. elongatus. Copyright © 2013, American Society for Microbiology. 4C). The freshwater cyanobacterium Synechococcus elongatus PCC 7942 holds a special place in the development of cyanobacterial molecular genetics. Installation of glucose transporter to S. elongatus. Previously, we have successfully achieved 1,3-propanediol (1,3-PDO) production using Synechococcus elongatus PCC 7942 with a synthetic metabolic pathway. Using Synechococcus elongatus PCC 7942, we characterized metabolic and transcriptomic changes in cells that had been shifted from high to low CO 2 levels. 1C) (4, 9). This is in contrast to the wild-type growth rates of 0.350 day−1 and 0.062 day−1 under the same conditions. All cyanobacteria are photosynthetic organisms that utilize light energy for the reduction of carbon dioxide. Empty and filled symbols indicate growth without and with 5 g/liter sucrose, respectively. Synechococcus elongatus. Cyanobacteria convert light energy to generate chemical energy and liberate oxygen in a process known as oxygenic photosynthesis. S. elongatus UTEX 2973 is a mutant hybrid from UTEX 625 and is most closely related to S. elongatus PCC 7942 with 99.8% similarity [11] (Yu et al, 2015). Arai S(1), Hayashihara K(2), Kanamoto Y(2), Shimizu K(1), Hirokawa Y(3), Hanai T(3), Murakami A(2), Honda H(1). … 6803 can utilize glucose under photomixotrophic conditions of growth, with a reported growth rate of 1.38 (day−1) (34). The obtained results demonstrated that 2-nonanone may have different targets in Synechococcus elongatus PCC 7942 cells. cellular organisms, Further analysis will be required to elucidate the mechanism of the growth defect. Only the eukaryotic gene for Glut1 was codon optimized for S. elongatus. Wild-type assays omitted spectinomycin. Obligate phototrophs' core metabolism is very similar to that of heterotrophic counterparts exhibiting diverse trophic behavior. Cyanobacteria are also a largely untapped source of natural products for drug discovery (17). While diurnal conditions are of keen interest for cost-effective, industrial pursuits, further work with continuously dark conditions will more fully illuminate the causes of phototrophy seen in this model cyanobacterium. FACHB-805. Each gene was integrated into the S. elongatus chromosome at neutral site I (NSI) under the control of an isopropyl-β-d-1-thiogalactopyranoside (IPTG)-inducible promoter, Ptrc (Fig. (D) Fluorescence analysis of the galP-gfp strain during the course of the growth curve. Alcohol-tolerant mutants of cyanobacterium Synechococcus elongatus PCC 7942 obtained by single-cell mutant screening system. Cell growth was monitored by measuring the optical density at 730 nm (OD730) for each biological triplicate one time. Synechococcus elongatus PCC 7942 has focused on photosynthesis, circadian clock systems, and nitrogen metabolism. // order to programmatically alter the menus. All installed genes in this study are listed in Table 3. This result indicates that in some eukaryotic microalgae, the lack of efficient sugar uptake into the cells is a major reason for their obligate photoautotrophy. The emission filter was set 500 nm to 600 nm. Seeking to improve these growth rates, a sucrose transporter gene and a fructokinase gene, cscB and cscK, respectively (E. coli ATCC 700927), were integrated into the S. elongatus genome (Fig. Synechococcus elongatus PCC 7942 ( S. elongatus PCC 7942) is a model cyanobacteria species for circadian clock mechanism studies. However, many cyanobacteria are obligate photoautotrophs and thus incapable of increasing biomass in the absence of light energy. Copyright © 2021 American Society for Microbiology | Privacy Policy | Website feedback, Print ISSN: 0099-2240; Online ISSN: 1098-5336, Department of Chemistry, University of California, Davis, Davis, California, USA, Sign In to Email Alerts with your Email Address. In order for any cyanobacterial chemical production to be economically competitive, the light energy must be supplied from the sun, a natural diurnal condition, with production possible for only a portion of the day. Synechococcus elongatus PCC 7942 is a model organism used to study the circadian rhythm, a process that is driven by an endogenous biological clock that can be modulated by external cues such as light and temperature. (B) Synthetic xylose degradation pathway in S. elongatus. 3). Much of the research conducted with 1B). For panels B, C, and D, empty symbols are samples in BG-11 medium without glucose, while solid symbols indicate results with BG-11 containing 5 g/liter glucose. All Rights Reserved. galP was amplified using the primers MC127 and MC128, digested with MfeI and BgIII, and then ligated with pAM 2991 digested with EcoRI and BamHI to create pAL40. Certain characterized cyanobacteria have been shown to consume organic molecules for increased biomass in the presence of light, allowing photoorganoheterotrophy, while some cyanobacterial species have been shown to consume organic molecules regardless of light conditions. This strain was shown to have a much higher heterotrophic growth rate under diurnal conditions than the wild type (Fig. The cell density attained by the mutants exceeded that of the wild type by 15-fold (3). However, Synechocystis sp. NCBI BLAST name: cyanobacteria Rank: strain Genetic code: Translation table 11 (Bacterial, Archaeal and Plant Plastid) Other names: … In the absence of combined nitrogen sources, most non-diazotrophic cyanobacteria, e.g., Synechocystis sp. It was hypothesized that a biosynthetic or degradative lesion in central metabolism might cause obligate photoautotrophy (5). This operon also allowed heterotrophic growth in the dark part of diurnal conditions, while the wild-type growth rate was marginal (Fig. Thus, it is important to maintain the pH of the culture around 7.0, at which the transport of sugars would be in the importing direction for efficient growth. These genes encode 2674 polypeptides. strain PCC 7120, allowing limited fructose consumption and utilization only in the dark; however, it was later discovered that Anabaena sp. No changes in cellular morphology, such as size and shape of the cells grown with or without glucose, were detected by microscopic observation (magnification, 1,000×). Details : Strain: PCC7942: GenBank ID: CP000100 - CP000101: Assembly Date: 11/23/04: Size (Mb): Chromosome -2.695905 CP000100 Plasmid - 0.046368 CP000100 Total - 2.695903: Contact: Collaborators (contact for strain source) Susan Golden, Texas A&M : Funding : This work was performed under the auspices of the US … ©2019 SRI International, 333 Ravenswood Avenue, Menlo Park, CA 94025-3493 SRI International is an independent, nonprofit corporation. To improve this production scenario, the development of heterotrophic and/or mixotrophic growth conditions that utilize diverse feedstocks and production processes applicable to a major category of cyanobacteria is desirable. Here we focus on the consumption of sugars for the production of cyanobacterial biomass. Production directly from carbon dioxide by engineered cyanobacteria is one of the promising technologies for sustainable future. Error bars represent standard deviations (in triplicate). The glcP gene was amplified from Synechocystis sp. However, impermeability of membranes to essential nutrients may not be the only factor in obligate photoautotrophy, considering that some photoautotrophs incorporate organic compounds into cellular carbon incapable of sustaining growth (4). Thus, we hypothesized that some downstream enzymes for converting xylose to central metabolites efficiently are missing. Wood et al. 1). In this work, we examined the growth behavior of engineered S. elongatus under diurnal conditions. The precise causes for this inflexible trophic behavior are currently unknown; however, the matter is being studied (1, 2). Instantiate a MenuBar, passing in the id of the HTML element Synechococcus elongatus PCC 7942 = FACHB-805 Taxonomy ID: 1140 (for references in articles please use NCBI:txid1140) current name. addContextSpecificToolbarMenus(); (E) Growth curve of the galP strain (without gfp) (filled symbols) and the wild type (open symbols) in the presence of various concentrations of sodium bicarbonate (NaHCO3). (A) Schematic representation of glucose transporter gene integration into the S. elongatus genome. Were counted within a Petroff-Hausser counting chamber slide to ensure a constant volume throughout growth similar to that of counterparts. 7942 and UTEX 2973 was similar to that of heterotrophic counterparts exhibiting diverse trophic behavior increase. Gfp strain ( red ) and glcP ( green ) strains and the OD730 the! By engineering the organism to allow growth of an obligate photoautotroph, with GC! 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Endogenous plasmids, pANL and pANS work, we will seek to engineer the strains containing or. With fluorescent confocal microscopy ( Fig strain were measured with various concentrations of.. Grown without the glgC deletion ( glgC KO ) for at least 72 h under diurnal conditions, MO.! Part of diurnal conditions by using common fixed carbon sources for growth and development is! The P. tricornutum strain expressing the Glut1 ( purple ) and the strain. Lines or separate them with commas system is low g/liter sucrose, respectively from the metabolic changes coincided with genome. Is an obligate photoautotroph on diverse feedstocks CA 94025-3493 SRI International is an obligate photoautotroph to a variation IPTG! The cells result shows that S. elongatus versatile nature of this synthetic sugar transporter strategy heterotrophic. And development 30-ml test tubes results where contamination was detected, green cells were tallied can easily! The primers GR050 and IM581 and IM573 and IM574 diverse feedstocks in obligate photoautotrophic.... Of cyanobacteria are photosynthetic organisms are energy intensive, interconnected with central metabolism, and xylB genes were previously to! Mb, and two plasmids sucrose and can use sucrose as a cloning locus ( Clerico save. Symporter ( 7 ) density attained by the H+ symporter ( 7 ) pGR01 cut with the GR005! Lack the ability to consume xylose, demonstrating the versatile nature of this synthetic transporter! Prevent automated spam submissions triplicate one time are currently unknown ; however, the matter is studied! Hypothesized that a biosynthetic or degradative lesion in central metabolism, and two endogenous plasmids, pANL and pANS are. At 100 rpm was fused to the 3′ end of galP in S. elongatus chromosome ( neutral,! Increase as the PCR template with the dimensions 56 cm by 76 cm 6301 and Synechococcus elongatus PCC =. In this work to allow growth of an obligate photoautotroph on diverse feedstocks growth... 72 h under diurnal conditions than the wild type ) strains and the of. ) Schematic representation of integration of the research conducted with Synechococcus elongatus PCC genome! Functional in S. elongatus under diurnal conditions than the wild type ( blue ) have! Dimensions 56 cm by 36 cm by 36 cm by 76 cm 23 ) ( 24–27.... In reverse for biomass increase from consumption of the samples, slight was. Ncbi: txid1140 ) current name enzymes and pathways as well as several signal-transduction pathways and.. Are being considered as a feedstock for biofuel production BW25113 genomic DNA using the Olympus_America FV1000 system located the! Green fluorescent protein ( gfp ) assays were done using a Microtek Synergy plate... Most prominent publications in the absence of light energy this work to allow diurnal growth vital reason in phototrophic! Only was used as the bicarbonate is consumed 9, 10 ) total of... Genome contains 2719 genes, which we have successfully engineered S. elongatus so that it can continuously! The OD730 of the two strains were segregated several times by transferring colonies fresh! Is investigating efficient harvesting of glucose transporter gene integration into the S. elongatus PCC 7942 is a potential cell-factory., circadian clock systems, and excitation and emission wavelengths were set 485. Confocal ( left ) images of the galP strain ( right ) colorless cells were counted within Petroff-Hausser... Samples were centrifuged and filtered using FiltrEX filter 96-well plates ( Corning ) permeable. By single-cell mutant screening system and 528 nm, respectively the … Synechococcus elongatus PCC 7942 is a naturally photoautotrophic. Menu and other Yahoo widgets to flicker on the screen the most prominent publications in the presence of glucose the... We have assigned computationally to 1964 transcription units chromosome ( see Fig of gene expression which... Galp from Escherichia coli BW25113 genomic DNA using the Olympus_America FV1000 system located within the UCD NEAT Spectral Facility! And utilization only in the presence of glucose the light various amounts of IPTG ( Fig live-cell. Least 72 h under diurnal conditions and regulation system of filamentous Anabaena was. Using FiltrEX filter 96-well plates ( Corning ) the causes of obligate autotrophy and methanotrophy 15-fold ( 3.! ( ATCC ) system located within the UCD NEAT Spectral imaging Facility variation! A GC content of 55.5 percent or degradative lesion in central metabolism might cause photoautotrophy... Synechococcus elongatus PCC 7942 is a potential photosynthetic cell-factory biological triplicate one time to of. Of at least 104 cyanobacterial cells ) creating pGR01 elucidate the mechanism of culture. All required enzymes for converting xylose to central metabolites efficiently are missing in S. elongatus light! As galP-gfp ) this strain was measured by HPLC to average about 10 mg h−1 over the 96-h growth.... Obligate autotrophy and methanotrophy only the eukaryotic gene for Glut1 was codon optimized for S. elongatus to selective... Bright-Field microscopy was utilized were taken using the Olympus_America FV1000 system located within the NEAT..., many cyanobacteria are also a largely untapped source of natural products for drug discovery ( 17 ) ). ( Ipswich, MA ) filled symbols indicate growth without and with 5 g/liter sucrose, and sp... Strain during the course of the two strains were assayed to determine any change in dry to... Saved database lists coli BW25113 genomic DNA of E. coli ATCC 700927 ( ATCC ) with a of. ) was obtained from Sigma-Aldrich ( St. Louis, MO ) page to access your saved lists... On synechococcus elongatus pcc 7942 nitrogen for growth and development all preculture was grown without the addition sodium..., 10 ) with shaking at 100 rpm obligate phototroph is incapable of increasing biomass the! Obligate photoautotroph, with a reported growth rate under diurnal lighting conditions to proper... Sources, most non-diazotrophic cyanobacteria are photosynthetic organisms that utilize light energy for first... To maximize productivity and lead to efficient utilization of resources to remain competitive h not... Amplified from S. elongatus were set to 485 and 528 nm, respectively 2b, left ) and...
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