acid 1 65 methionine  (ATCC)


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    ATCC acid 1 65 methionine
    Acid 1 65 Methionine, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    methionine  (ATCC)


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    ATCC methionine
    C. amycolatum WT, C. amycolatum cob-, C. glucuronolyticum, C. glutamicum, and C. kefirresidentii were cultured in minimal cobamide-free medium with 0, 50, or 250 nM CoCl 2 concentrations. Intracellular and extracellular cobamides were estimated using an E. coli microbiological indicator assay . Cobamide vs. <t>methionine</t> presence in the samples was confirmed using both the E. coli metE- and E. coli metE - Δ metH indicator strains. Intracellular cobamides were quantified per gram of wet cell weight, and extracellular (CFSM) cobamides were quantified per mL of media. Intracellular cobamides were converted to their cyano form before testing in the indicator assay, and extracellular cobamides were tested in their native form.
    Methionine, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Skin-associated Corynebacterium amycolatum shares cobamides"

    Article Title: Skin-associated Corynebacterium amycolatum shares cobamides

    Journal: bioRxiv

    doi: 10.1101/2024.04.28.591522

    C. amycolatum WT, C. amycolatum cob-, C. glucuronolyticum, C. glutamicum, and C. kefirresidentii were cultured in minimal cobamide-free medium with 0, 50, or 250 nM CoCl 2 concentrations. Intracellular and extracellular cobamides were estimated using an E. coli microbiological indicator assay . Cobamide vs. methionine presence in the samples was confirmed using both the E. coli metE- and E. coli metE - Δ metH indicator strains. Intracellular cobamides were quantified per gram of wet cell weight, and extracellular (CFSM) cobamides were quantified per mL of media. Intracellular cobamides were converted to their cyano form before testing in the indicator assay, and extracellular cobamides were tested in their native form.
    Figure Legend Snippet: C. amycolatum WT, C. amycolatum cob-, C. glucuronolyticum, C. glutamicum, and C. kefirresidentii were cultured in minimal cobamide-free medium with 0, 50, or 250 nM CoCl 2 concentrations. Intracellular and extracellular cobamides were estimated using an E. coli microbiological indicator assay . Cobamide vs. methionine presence in the samples was confirmed using both the E. coli metE- and E. coli metE - Δ metH indicator strains. Intracellular cobamides were quantified per gram of wet cell weight, and extracellular (CFSM) cobamides were quantified per mL of media. Intracellular cobamides were converted to their cyano form before testing in the indicator assay, and extracellular cobamides were tested in their native form.

    Techniques Used: Cell Culture

    (A) E. coli metE - (blue) was cultured with 0, 1, and 2.5 ng/mL cyanocobalamin standards (CnCbl), and (B) E. coli metE - Δ metH (pink) was cultured with 0, 10, and 100 μg/mL methionine to validate expected growth response of each strain, quantified in CFU/mL. Growth of E. coli metE - is proportional to the concentration of cobamides or methionine in the medium, and growth of E. coli metE - Δ metH is proportional to the concentration of only methionine, thus distinguishing between cobamides and methionine in supporting E. coli growth. C. amycolatum WT (WT) was co-cultured in minimal medium with (C) E. coli metE - (blue) or (D) E. coli metE - Δ metH (pink) under low (0 nM) or high (250 nM) cobalt concentrations. C. amycolatum cob - (cob-) was co-cultured in minimal medium with (E) E. coli metE - (blue) or (F) E. coli metE - Δ metH (pink) under low (0 nM) or high (250 nM) cobalt concentrations. (G) C. amycolatum CFU/mL was measured to demonstrate growth across each strain condition. n=9 replicates for each condition (3 technical replicates across 3 biological replicates). Solid lines represent the measured inoculum for E. coli metE - , E. coli metE - Δ metH , or C. amycolatum averaged across all replicates. FS = filter-sterilized C. amycolatum cell suspension, L = lysed C. amycolatum cell suspension.
    Figure Legend Snippet: (A) E. coli metE - (blue) was cultured with 0, 1, and 2.5 ng/mL cyanocobalamin standards (CnCbl), and (B) E. coli metE - Δ metH (pink) was cultured with 0, 10, and 100 μg/mL methionine to validate expected growth response of each strain, quantified in CFU/mL. Growth of E. coli metE - is proportional to the concentration of cobamides or methionine in the medium, and growth of E. coli metE - Δ metH is proportional to the concentration of only methionine, thus distinguishing between cobamides and methionine in supporting E. coli growth. C. amycolatum WT (WT) was co-cultured in minimal medium with (C) E. coli metE - (blue) or (D) E. coli metE - Δ metH (pink) under low (0 nM) or high (250 nM) cobalt concentrations. C. amycolatum cob - (cob-) was co-cultured in minimal medium with (E) E. coli metE - (blue) or (F) E. coli metE - Δ metH (pink) under low (0 nM) or high (250 nM) cobalt concentrations. (G) C. amycolatum CFU/mL was measured to demonstrate growth across each strain condition. n=9 replicates for each condition (3 technical replicates across 3 biological replicates). Solid lines represent the measured inoculum for E. coli metE - , E. coli metE - Δ metH , or C. amycolatum averaged across all replicates. FS = filter-sterilized C. amycolatum cell suspension, L = lysed C. amycolatum cell suspension.

    Techniques Used: Cell Culture, Concentration Assay, Suspension

    C. amycolatum WT or cob - cell suspension or controls (CnCbl, methionine, filtered cell suspension (FS), lysed cell suspension (L) were added to half of a minimal medium plate and incubated for 3 days, after which E. coli metE - (blue) or E. coli metE - Δ metH (pink) was struck out on the adjacent side of the plate. E. coli growth was recorded after 24 h. Growth summary for all conditions is indicated in (A). - indicates no growth and +, ++, +++ indicate increasing levels of growth, respectively. C. amycolatum WT lawn with (B) E. coli metE - and (C) E. coli metE - Δ metH. C. amycolatum cob - lawn with (D) E. coli metE - , (E) E. coli metE - + 100 ng/mL CnCbl, and (F) E. coli metE - Δ metH. E. coli metE - with (G) C. amycolatum filter-sterilized cell suspension and (H) C. amycolatum lysed cell suspension. Blank controls for (I) E. coli metE - and (J) E. coli metE - Δ metH . (K) E. coli metE - and (L) E. coli metE - Δ metH growth response to CnCbl and methionine standards.
    Figure Legend Snippet: C. amycolatum WT or cob - cell suspension or controls (CnCbl, methionine, filtered cell suspension (FS), lysed cell suspension (L) were added to half of a minimal medium plate and incubated for 3 days, after which E. coli metE - (blue) or E. coli metE - Δ metH (pink) was struck out on the adjacent side of the plate. E. coli growth was recorded after 24 h. Growth summary for all conditions is indicated in (A). - indicates no growth and +, ++, +++ indicate increasing levels of growth, respectively. C. amycolatum WT lawn with (B) E. coli metE - and (C) E. coli metE - Δ metH. C. amycolatum cob - lawn with (D) E. coli metE - , (E) E. coli metE - + 100 ng/mL CnCbl, and (F) E. coli metE - Δ metH. E. coli metE - with (G) C. amycolatum filter-sterilized cell suspension and (H) C. amycolatum lysed cell suspension. Blank controls for (I) E. coli metE - and (J) E. coli metE - Δ metH . (K) E. coli metE - and (L) E. coli metE - Δ metH growth response to CnCbl and methionine standards.

    Techniques Used: Suspension, Incubation

    methionine synthase meth  (ATCC)


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    ATCC methionine synthase meth
    C. amycolatum WT, C. amycolatum cob-, C. glucuronolyticum, C. glutamicum, and C. kefirresidentii were cultured in minimal cobamide-free medium with 0, 50, or 250 nM CoCl 2 concentrations. Intracellular and extracellular cobamides were estimated using an E. coli microbiological indicator assay . Cobamide vs. <t>methionine</t> presence in the samples was confirmed using both the E. coli metE- and E. coli metE - Δ <t>metH</t> indicator strains. Intracellular cobamides were quantified per gram of wet cell weight, and extracellular (CFSM) cobamides were quantified per mL of media. Intracellular cobamides were converted to their cyano form before testing in the indicator assay, and extracellular cobamides were tested in their native form.
    Methionine Synthase Meth, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Skin-associated Corynebacterium amycolatum shares cobamides"

    Article Title: Skin-associated Corynebacterium amycolatum shares cobamides

    Journal: bioRxiv

    doi: 10.1101/2024.04.28.591522

    C. amycolatum WT, C. amycolatum cob-, C. glucuronolyticum, C. glutamicum, and C. kefirresidentii were cultured in minimal cobamide-free medium with 0, 50, or 250 nM CoCl 2 concentrations. Intracellular and extracellular cobamides were estimated using an E. coli microbiological indicator assay . Cobamide vs. methionine presence in the samples was confirmed using both the E. coli metE- and E. coli metE - Δ metH indicator strains. Intracellular cobamides were quantified per gram of wet cell weight, and extracellular (CFSM) cobamides were quantified per mL of media. Intracellular cobamides were converted to their cyano form before testing in the indicator assay, and extracellular cobamides were tested in their native form.
    Figure Legend Snippet: C. amycolatum WT, C. amycolatum cob-, C. glucuronolyticum, C. glutamicum, and C. kefirresidentii were cultured in minimal cobamide-free medium with 0, 50, or 250 nM CoCl 2 concentrations. Intracellular and extracellular cobamides were estimated using an E. coli microbiological indicator assay . Cobamide vs. methionine presence in the samples was confirmed using both the E. coli metE- and E. coli metE - Δ metH indicator strains. Intracellular cobamides were quantified per gram of wet cell weight, and extracellular (CFSM) cobamides were quantified per mL of media. Intracellular cobamides were converted to their cyano form before testing in the indicator assay, and extracellular cobamides were tested in their native form.

    Techniques Used: Cell Culture

    (A) E. coli metE - (blue) was cultured with 0, 1, and 2.5 ng/mL cyanocobalamin standards (CnCbl), and (B) E. coli metE - Δ metH (pink) was cultured with 0, 10, and 100 μg/mL methionine to validate expected growth response of each strain, quantified in CFU/mL. Growth of E. coli metE - is proportional to the concentration of cobamides or methionine in the medium, and growth of E. coli metE - Δ metH is proportional to the concentration of only methionine, thus distinguishing between cobamides and methionine in supporting E. coli growth. C. amycolatum WT (WT) was co-cultured in minimal medium with (C) E. coli metE - (blue) or (D) E. coli metE - Δ metH (pink) under low (0 nM) or high (250 nM) cobalt concentrations. C. amycolatum cob - (cob-) was co-cultured in minimal medium with (E) E. coli metE - (blue) or (F) E. coli metE - Δ metH (pink) under low (0 nM) or high (250 nM) cobalt concentrations. (G) C. amycolatum CFU/mL was measured to demonstrate growth across each strain condition. n=9 replicates for each condition (3 technical replicates across 3 biological replicates). Solid lines represent the measured inoculum for E. coli metE - , E. coli metE - Δ metH , or C. amycolatum averaged across all replicates. FS = filter-sterilized C. amycolatum cell suspension, L = lysed C. amycolatum cell suspension.
    Figure Legend Snippet: (A) E. coli metE - (blue) was cultured with 0, 1, and 2.5 ng/mL cyanocobalamin standards (CnCbl), and (B) E. coli metE - Δ metH (pink) was cultured with 0, 10, and 100 μg/mL methionine to validate expected growth response of each strain, quantified in CFU/mL. Growth of E. coli metE - is proportional to the concentration of cobamides or methionine in the medium, and growth of E. coli metE - Δ metH is proportional to the concentration of only methionine, thus distinguishing between cobamides and methionine in supporting E. coli growth. C. amycolatum WT (WT) was co-cultured in minimal medium with (C) E. coli metE - (blue) or (D) E. coli metE - Δ metH (pink) under low (0 nM) or high (250 nM) cobalt concentrations. C. amycolatum cob - (cob-) was co-cultured in minimal medium with (E) E. coli metE - (blue) or (F) E. coli metE - Δ metH (pink) under low (0 nM) or high (250 nM) cobalt concentrations. (G) C. amycolatum CFU/mL was measured to demonstrate growth across each strain condition. n=9 replicates for each condition (3 technical replicates across 3 biological replicates). Solid lines represent the measured inoculum for E. coli metE - , E. coli metE - Δ metH , or C. amycolatum averaged across all replicates. FS = filter-sterilized C. amycolatum cell suspension, L = lysed C. amycolatum cell suspension.

    Techniques Used: Cell Culture, Concentration Assay, Suspension

    C. amycolatum WT or cob - cell suspension or controls (CnCbl, methionine, filtered cell suspension (FS), lysed cell suspension (L) were added to half of a minimal medium plate and incubated for 3 days, after which E. coli metE - (blue) or E. coli metE - Δ metH (pink) was struck out on the adjacent side of the plate. E. coli growth was recorded after 24 h. Growth summary for all conditions is indicated in (A). - indicates no growth and +, ++, +++ indicate increasing levels of growth, respectively. C. amycolatum WT lawn with (B) E. coli metE - and (C) E. coli metE - Δ metH. C. amycolatum cob - lawn with (D) E. coli metE - , (E) E. coli metE - + 100 ng/mL CnCbl, and (F) E. coli metE - Δ metH. E. coli metE - with (G) C. amycolatum filter-sterilized cell suspension and (H) C. amycolatum lysed cell suspension. Blank controls for (I) E. coli metE - and (J) E. coli metE - Δ metH . (K) E. coli metE - and (L) E. coli metE - Δ metH growth response to CnCbl and methionine standards.
    Figure Legend Snippet: C. amycolatum WT or cob - cell suspension or controls (CnCbl, methionine, filtered cell suspension (FS), lysed cell suspension (L) were added to half of a minimal medium plate and incubated for 3 days, after which E. coli metE - (blue) or E. coli metE - Δ metH (pink) was struck out on the adjacent side of the plate. E. coli growth was recorded after 24 h. Growth summary for all conditions is indicated in (A). - indicates no growth and +, ++, +++ indicate increasing levels of growth, respectively. C. amycolatum WT lawn with (B) E. coli metE - and (C) E. coli metE - Δ metH. C. amycolatum cob - lawn with (D) E. coli metE - , (E) E. coli metE - + 100 ng/mL CnCbl, and (F) E. coli metE - Δ metH. E. coli metE - with (G) C. amycolatum filter-sterilized cell suspension and (H) C. amycolatum lysed cell suspension. Blank controls for (I) E. coli metE - and (J) E. coli metE - Δ metH . (K) E. coli metE - and (L) E. coli metE - Δ metH growth response to CnCbl and methionine standards.

    Techniques Used: Suspension, Incubation

    methionine  (ATCC)


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    ATCC methionine
    Methionine, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    methionine synthesis  (ATCC)


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    ATCC methionine synthesis
    Methionine Synthesis, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    methionine salvage  (ATCC)


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    ATCC methionine salvage
    Methionine Salvage, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    methionine salvage pathways  (ATCC)


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    ATCC methionine salvage pathways
    Methionine Salvage Pathways, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    methionine synthesis  (ATCC)


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    ATCC methionine synthesis
    Salvage of S-adenosyl- l <t>-methionine</t> (SAM) utilization byproducts, 5-deoxy-pentose sugars, and 6-deoxy-hexose sugars in E. coli . ( A ) S -adenosyl- l -homocysteine (SAH), produced by methyltransferases, is recycled by the methionine cycle (a.k.a. active methyl cycle) ( , ). ( B and C ) The E. coli variation of the DHAP shunt. ( B ) For adenine salvage and detoxification of 5′-methylthioadenosine (MTA) and 5′-deoxyadenosine (5dAdo), all E. coli possess the multifunctional SAH/MTA/5dAdo nucleosidase (Pfs; a.k.a. MtnN) . ( C ) ExPEC strains possess the Pfs nucleosidase ( B ) and remainder of the DHAP shunt genes, which together compose a dual-purpose pathway for the conversion of 5′-deoxy-nucleosides in the form of MTA and 5dAdo or corresponding 5-deoxy-pentoses in the form of 5-deoxy- d -ribose and 5-methylthioribose, respectively, into the central carbon metabolite DHAP and acetaldehyde or (2-methylthio)acetaldehyde . ( D ) E. coli analogously metabolize 6-deoxy-hexose sugars in the form of l -fucose and l -rhamnose to DHAP and l -lactaldehyde. During anaerobic growth, l -lactaldehyde is primarily reduced to ( S )-1,2-propanediol as a terminal product, whereas during aerobic growth, it is primarily oxidized to l -lactate for carbon assimilation as pyruvate. Enzymes: Pfs (MtnN), SAH/MTA/5dAdo nucleosidase, E.C. 3.2.2.9; LuxS, S -ribosyl- l -homocysteine lyase, E.C. 4.4.1.21; MetH, methionine synthase, E.C. 2.1.1.13; MetE, methionine synthase, E.C. 2.1.1.4; MtnK, 5-methylthioribose/5-deoxyribose kinase, E.C. 2.7.1.100; MtnA, 5-methylthioribose-1-phosphate/5-deoxyribose-1-phosphate isomerase, E.C. 5.3.1.23; Ald2, 5-methylthioribulose-1-phosphate/5-deoxyribulose-1-phosphate aldolase, E.C. 4.1.2.62; ADH, alcohol dehydrogenase, E.C. 1.1.1.1 ; FucI, l -fucose isomerase, E.C. 5.3.1.25; FucK, l -fuculose kinase, E.C. 2.7.1.51; FucA, l -fuculose-1-phosphate aldolase, E.C. 4.1.2.17; RhaA, l -rhamnose isomerase, E.C. 5.3.1.14; RhaB, l -rhamnulose kinase, E.C. 2.7.1.5; RhaD, l -rhamnulose-1-phosphate aldolase, E.C. 4.1.2.19; FucO, ( S )−1,2 -propanediol oxidoreductase, E.C. 1.1.1.77; and AldA, l -lactaldehyde dehydrogenase, E.C. 1.2.1.22.
    Methionine Synthesis, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Escherichia coli possessing the dihydroxyacetone phosphate shunt utilize 5′-deoxynucleosides for growth"

    Article Title: Escherichia coli possessing the dihydroxyacetone phosphate shunt utilize 5′-deoxynucleosides for growth

    Journal: Microbiology Spectrum

    doi: 10.1128/spectrum.03086-23

    Salvage of S-adenosyl- l -methionine (SAM) utilization byproducts, 5-deoxy-pentose sugars, and 6-deoxy-hexose sugars in E. coli . ( A ) S -adenosyl- l -homocysteine (SAH), produced by methyltransferases, is recycled by the methionine cycle (a.k.a. active methyl cycle) ( , ). ( B and C ) The E. coli variation of the DHAP shunt. ( B ) For adenine salvage and detoxification of 5′-methylthioadenosine (MTA) and 5′-deoxyadenosine (5dAdo), all E. coli possess the multifunctional SAH/MTA/5dAdo nucleosidase (Pfs; a.k.a. MtnN) . ( C ) ExPEC strains possess the Pfs nucleosidase ( B ) and remainder of the DHAP shunt genes, which together compose a dual-purpose pathway for the conversion of 5′-deoxy-nucleosides in the form of MTA and 5dAdo or corresponding 5-deoxy-pentoses in the form of 5-deoxy- d -ribose and 5-methylthioribose, respectively, into the central carbon metabolite DHAP and acetaldehyde or (2-methylthio)acetaldehyde . ( D ) E. coli analogously metabolize 6-deoxy-hexose sugars in the form of l -fucose and l -rhamnose to DHAP and l -lactaldehyde. During anaerobic growth, l -lactaldehyde is primarily reduced to ( S )-1,2-propanediol as a terminal product, whereas during aerobic growth, it is primarily oxidized to l -lactate for carbon assimilation as pyruvate. Enzymes: Pfs (MtnN), SAH/MTA/5dAdo nucleosidase, E.C. 3.2.2.9; LuxS, S -ribosyl- l -homocysteine lyase, E.C. 4.4.1.21; MetH, methionine synthase, E.C. 2.1.1.13; MetE, methionine synthase, E.C. 2.1.1.4; MtnK, 5-methylthioribose/5-deoxyribose kinase, E.C. 2.7.1.100; MtnA, 5-methylthioribose-1-phosphate/5-deoxyribose-1-phosphate isomerase, E.C. 5.3.1.23; Ald2, 5-methylthioribulose-1-phosphate/5-deoxyribulose-1-phosphate aldolase, E.C. 4.1.2.62; ADH, alcohol dehydrogenase, E.C. 1.1.1.1 ; FucI, l -fucose isomerase, E.C. 5.3.1.25; FucK, l -fuculose kinase, E.C. 2.7.1.51; FucA, l -fuculose-1-phosphate aldolase, E.C. 4.1.2.17; RhaA, l -rhamnose isomerase, E.C. 5.3.1.14; RhaB, l -rhamnulose kinase, E.C. 2.7.1.5; RhaD, l -rhamnulose-1-phosphate aldolase, E.C. 4.1.2.19; FucO, ( S )−1,2 -propanediol oxidoreductase, E.C. 1.1.1.77; and AldA, l -lactaldehyde dehydrogenase, E.C. 1.2.1.22.
    Figure Legend Snippet: Salvage of S-adenosyl- l -methionine (SAM) utilization byproducts, 5-deoxy-pentose sugars, and 6-deoxy-hexose sugars in E. coli . ( A ) S -adenosyl- l -homocysteine (SAH), produced by methyltransferases, is recycled by the methionine cycle (a.k.a. active methyl cycle) ( , ). ( B and C ) The E. coli variation of the DHAP shunt. ( B ) For adenine salvage and detoxification of 5′-methylthioadenosine (MTA) and 5′-deoxyadenosine (5dAdo), all E. coli possess the multifunctional SAH/MTA/5dAdo nucleosidase (Pfs; a.k.a. MtnN) . ( C ) ExPEC strains possess the Pfs nucleosidase ( B ) and remainder of the DHAP shunt genes, which together compose a dual-purpose pathway for the conversion of 5′-deoxy-nucleosides in the form of MTA and 5dAdo or corresponding 5-deoxy-pentoses in the form of 5-deoxy- d -ribose and 5-methylthioribose, respectively, into the central carbon metabolite DHAP and acetaldehyde or (2-methylthio)acetaldehyde . ( D ) E. coli analogously metabolize 6-deoxy-hexose sugars in the form of l -fucose and l -rhamnose to DHAP and l -lactaldehyde. During anaerobic growth, l -lactaldehyde is primarily reduced to ( S )-1,2-propanediol as a terminal product, whereas during aerobic growth, it is primarily oxidized to l -lactate for carbon assimilation as pyruvate. Enzymes: Pfs (MtnN), SAH/MTA/5dAdo nucleosidase, E.C. 3.2.2.9; LuxS, S -ribosyl- l -homocysteine lyase, E.C. 4.4.1.21; MetH, methionine synthase, E.C. 2.1.1.13; MetE, methionine synthase, E.C. 2.1.1.4; MtnK, 5-methylthioribose/5-deoxyribose kinase, E.C. 2.7.1.100; MtnA, 5-methylthioribose-1-phosphate/5-deoxyribose-1-phosphate isomerase, E.C. 5.3.1.23; Ald2, 5-methylthioribulose-1-phosphate/5-deoxyribulose-1-phosphate aldolase, E.C. 4.1.2.62; ADH, alcohol dehydrogenase, E.C. 1.1.1.1 ; FucI, l -fucose isomerase, E.C. 5.3.1.25; FucK, l -fuculose kinase, E.C. 2.7.1.51; FucA, l -fuculose-1-phosphate aldolase, E.C. 4.1.2.17; RhaA, l -rhamnose isomerase, E.C. 5.3.1.14; RhaB, l -rhamnulose kinase, E.C. 2.7.1.5; RhaD, l -rhamnulose-1-phosphate aldolase, E.C. 4.1.2.19; FucO, ( S )−1,2 -propanediol oxidoreductase, E.C. 1.1.1.77; and AldA, l -lactaldehyde dehydrogenase, E.C. 1.2.1.22.

    Techniques Used: Produced

    Sulfur from MTA cannot be salvaged by E. coli for growth. ( A ) ATCC 25922 maximum growth achieved after 24 hours measured by optical density at 600 nm when cultured with 1 mM of the indicated sulfur compound as the sole sulfur source. No further growth was observed after 24 hours. Average and standard deviation error bars are for n = 3 independent replicates. ( B ) Fold difference in the abundance of DHAP shunt-associated metabolites when ATCC 25922 was grown aerobically in the presence of 1 mM sulfate and 1 mM MTA versus grown aerobically in the presence of 1 mM sulfate only. Metabolites were resolved by LC-MS/MS from three independent biological replicates for each growth condition. Values are the average for the three replicates, and the significance of fold change was analyzed by ANOVA with P < 0.05. ( C ) Reverse-phase HPLC quantification of E. coli ATCC 25922 metabolites when fed aerobically with [ 14 C-methyl]−5′-methylthioadenosine. Unk, unknown (identified as 2-methylsulfinylethanol by LC-MS/MS, m / z = 109.0319), R T = 6.7 min; Met, methionine, R T = 8.0 min; MTR, methylthioribose, R T = 16.7 min; MT-EtOH, 2-methylthioethanol, R T = 21.7 min; IS, internal standard, R T = 24.0 min; and MTA, 5′-methylthioadenosine, R T = 31.8 min.
    Figure Legend Snippet: Sulfur from MTA cannot be salvaged by E. coli for growth. ( A ) ATCC 25922 maximum growth achieved after 24 hours measured by optical density at 600 nm when cultured with 1 mM of the indicated sulfur compound as the sole sulfur source. No further growth was observed after 24 hours. Average and standard deviation error bars are for n = 3 independent replicates. ( B ) Fold difference in the abundance of DHAP shunt-associated metabolites when ATCC 25922 was grown aerobically in the presence of 1 mM sulfate and 1 mM MTA versus grown aerobically in the presence of 1 mM sulfate only. Metabolites were resolved by LC-MS/MS from three independent biological replicates for each growth condition. Values are the average for the three replicates, and the significance of fold change was analyzed by ANOVA with P < 0.05. ( C ) Reverse-phase HPLC quantification of E. coli ATCC 25922 metabolites when fed aerobically with [ 14 C-methyl]−5′-methylthioadenosine. Unk, unknown (identified as 2-methylsulfinylethanol by LC-MS/MS, m / z = 109.0319), R T = 6.7 min; Met, methionine, R T = 8.0 min; MTR, methylthioribose, R T = 16.7 min; MT-EtOH, 2-methylthioethanol, R T = 21.7 min; IS, internal standard, R T = 24.0 min; and MTA, 5′-methylthioadenosine, R T = 31.8 min.

    Techniques Used: Cell Culture, Standard Deviation, Liquid Chromatography with Mass Spectroscopy

    methionine  (ATCC)


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    Structured Review

    ATCC methionine
    Salvage of S-adenosyl- l <t>-methionine</t> (SAM) utilization byproducts, 5-deoxy-pentose sugars, and 6-deoxy-hexose sugars in E. coli . ( A ) S -adenosyl- l -homocysteine (SAH), produced by methyltransferases, is recycled by the methionine cycle (a.k.a. active methyl cycle) ( , ). ( B and C ) The E. coli variation of the DHAP shunt. ( B ) For adenine salvage and detoxification of 5′-methylthioadenosine (MTA) and 5′-deoxyadenosine (5dAdo), all E. coli possess the multifunctional SAH/MTA/5dAdo nucleosidase (Pfs; a.k.a. MtnN) . ( C ) ExPEC strains possess the Pfs nucleosidase ( B ) and remainder of the DHAP shunt genes, which together compose a dual-purpose pathway for the conversion of 5′-deoxy-nucleosides in the form of MTA and 5dAdo or corresponding 5-deoxy-pentoses in the form of 5-deoxy- d -ribose and 5-methylthioribose, respectively, into the central carbon metabolite DHAP and acetaldehyde or (2-methylthio)acetaldehyde . ( D ) E. coli analogously metabolize 6-deoxy-hexose sugars in the form of l -fucose and l -rhamnose to DHAP and l -lactaldehyde. During anaerobic growth, l -lactaldehyde is primarily reduced to ( S )-1,2-propanediol as a terminal product, whereas during aerobic growth, it is primarily oxidized to l -lactate for carbon assimilation as pyruvate. Enzymes: Pfs (MtnN), SAH/MTA/5dAdo nucleosidase, E.C. 3.2.2.9; LuxS, S -ribosyl- l -homocysteine lyase, E.C. 4.4.1.21; MetH, methionine synthase, E.C. 2.1.1.13; MetE, methionine synthase, E.C. 2.1.1.4; MtnK, 5-methylthioribose/5-deoxyribose kinase, E.C. 2.7.1.100; MtnA, 5-methylthioribose-1-phosphate/5-deoxyribose-1-phosphate isomerase, E.C. 5.3.1.23; Ald2, 5-methylthioribulose-1-phosphate/5-deoxyribulose-1-phosphate aldolase, E.C. 4.1.2.62; ADH, alcohol dehydrogenase, E.C. 1.1.1.1 ; FucI, l -fucose isomerase, E.C. 5.3.1.25; FucK, l -fuculose kinase, E.C. 2.7.1.51; FucA, l -fuculose-1-phosphate aldolase, E.C. 4.1.2.17; RhaA, l -rhamnose isomerase, E.C. 5.3.1.14; RhaB, l -rhamnulose kinase, E.C. 2.7.1.5; RhaD, l -rhamnulose-1-phosphate aldolase, E.C. 4.1.2.19; FucO, ( S )−1,2 -propanediol oxidoreductase, E.C. 1.1.1.77; and AldA, l -lactaldehyde dehydrogenase, E.C. 1.2.1.22.
    Methionine, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Escherichia coli possessing the dihydroxyacetone phosphate shunt utilize 5′-deoxynucleosides for growth"

    Article Title: Escherichia coli possessing the dihydroxyacetone phosphate shunt utilize 5′-deoxynucleosides for growth

    Journal: Microbiology Spectrum

    doi: 10.1128/spectrum.03086-23

    Salvage of S-adenosyl- l -methionine (SAM) utilization byproducts, 5-deoxy-pentose sugars, and 6-deoxy-hexose sugars in E. coli . ( A ) S -adenosyl- l -homocysteine (SAH), produced by methyltransferases, is recycled by the methionine cycle (a.k.a. active methyl cycle) ( , ). ( B and C ) The E. coli variation of the DHAP shunt. ( B ) For adenine salvage and detoxification of 5′-methylthioadenosine (MTA) and 5′-deoxyadenosine (5dAdo), all E. coli possess the multifunctional SAH/MTA/5dAdo nucleosidase (Pfs; a.k.a. MtnN) . ( C ) ExPEC strains possess the Pfs nucleosidase ( B ) and remainder of the DHAP shunt genes, which together compose a dual-purpose pathway for the conversion of 5′-deoxy-nucleosides in the form of MTA and 5dAdo or corresponding 5-deoxy-pentoses in the form of 5-deoxy- d -ribose and 5-methylthioribose, respectively, into the central carbon metabolite DHAP and acetaldehyde or (2-methylthio)acetaldehyde . ( D ) E. coli analogously metabolize 6-deoxy-hexose sugars in the form of l -fucose and l -rhamnose to DHAP and l -lactaldehyde. During anaerobic growth, l -lactaldehyde is primarily reduced to ( S )-1,2-propanediol as a terminal product, whereas during aerobic growth, it is primarily oxidized to l -lactate for carbon assimilation as pyruvate. Enzymes: Pfs (MtnN), SAH/MTA/5dAdo nucleosidase, E.C. 3.2.2.9; LuxS, S -ribosyl- l -homocysteine lyase, E.C. 4.4.1.21; MetH, methionine synthase, E.C. 2.1.1.13; MetE, methionine synthase, E.C. 2.1.1.4; MtnK, 5-methylthioribose/5-deoxyribose kinase, E.C. 2.7.1.100; MtnA, 5-methylthioribose-1-phosphate/5-deoxyribose-1-phosphate isomerase, E.C. 5.3.1.23; Ald2, 5-methylthioribulose-1-phosphate/5-deoxyribulose-1-phosphate aldolase, E.C. 4.1.2.62; ADH, alcohol dehydrogenase, E.C. 1.1.1.1 ; FucI, l -fucose isomerase, E.C. 5.3.1.25; FucK, l -fuculose kinase, E.C. 2.7.1.51; FucA, l -fuculose-1-phosphate aldolase, E.C. 4.1.2.17; RhaA, l -rhamnose isomerase, E.C. 5.3.1.14; RhaB, l -rhamnulose kinase, E.C. 2.7.1.5; RhaD, l -rhamnulose-1-phosphate aldolase, E.C. 4.1.2.19; FucO, ( S )−1,2 -propanediol oxidoreductase, E.C. 1.1.1.77; and AldA, l -lactaldehyde dehydrogenase, E.C. 1.2.1.22.
    Figure Legend Snippet: Salvage of S-adenosyl- l -methionine (SAM) utilization byproducts, 5-deoxy-pentose sugars, and 6-deoxy-hexose sugars in E. coli . ( A ) S -adenosyl- l -homocysteine (SAH), produced by methyltransferases, is recycled by the methionine cycle (a.k.a. active methyl cycle) ( , ). ( B and C ) The E. coli variation of the DHAP shunt. ( B ) For adenine salvage and detoxification of 5′-methylthioadenosine (MTA) and 5′-deoxyadenosine (5dAdo), all E. coli possess the multifunctional SAH/MTA/5dAdo nucleosidase (Pfs; a.k.a. MtnN) . ( C ) ExPEC strains possess the Pfs nucleosidase ( B ) and remainder of the DHAP shunt genes, which together compose a dual-purpose pathway for the conversion of 5′-deoxy-nucleosides in the form of MTA and 5dAdo or corresponding 5-deoxy-pentoses in the form of 5-deoxy- d -ribose and 5-methylthioribose, respectively, into the central carbon metabolite DHAP and acetaldehyde or (2-methylthio)acetaldehyde . ( D ) E. coli analogously metabolize 6-deoxy-hexose sugars in the form of l -fucose and l -rhamnose to DHAP and l -lactaldehyde. During anaerobic growth, l -lactaldehyde is primarily reduced to ( S )-1,2-propanediol as a terminal product, whereas during aerobic growth, it is primarily oxidized to l -lactate for carbon assimilation as pyruvate. Enzymes: Pfs (MtnN), SAH/MTA/5dAdo nucleosidase, E.C. 3.2.2.9; LuxS, S -ribosyl- l -homocysteine lyase, E.C. 4.4.1.21; MetH, methionine synthase, E.C. 2.1.1.13; MetE, methionine synthase, E.C. 2.1.1.4; MtnK, 5-methylthioribose/5-deoxyribose kinase, E.C. 2.7.1.100; MtnA, 5-methylthioribose-1-phosphate/5-deoxyribose-1-phosphate isomerase, E.C. 5.3.1.23; Ald2, 5-methylthioribulose-1-phosphate/5-deoxyribulose-1-phosphate aldolase, E.C. 4.1.2.62; ADH, alcohol dehydrogenase, E.C. 1.1.1.1 ; FucI, l -fucose isomerase, E.C. 5.3.1.25; FucK, l -fuculose kinase, E.C. 2.7.1.51; FucA, l -fuculose-1-phosphate aldolase, E.C. 4.1.2.17; RhaA, l -rhamnose isomerase, E.C. 5.3.1.14; RhaB, l -rhamnulose kinase, E.C. 2.7.1.5; RhaD, l -rhamnulose-1-phosphate aldolase, E.C. 4.1.2.19; FucO, ( S )−1,2 -propanediol oxidoreductase, E.C. 1.1.1.77; and AldA, l -lactaldehyde dehydrogenase, E.C. 1.2.1.22.

    Techniques Used: Produced

    Sulfur from MTA cannot be salvaged by E. coli for growth. ( A ) ATCC 25922 maximum growth achieved after 24 hours measured by optical density at 600 nm when cultured with 1 mM of the indicated sulfur compound as the sole sulfur source. No further growth was observed after 24 hours. Average and standard deviation error bars are for n = 3 independent replicates. ( B ) Fold difference in the abundance of DHAP shunt-associated metabolites when ATCC 25922 was grown aerobically in the presence of 1 mM sulfate and 1 mM MTA versus grown aerobically in the presence of 1 mM sulfate only. Metabolites were resolved by LC-MS/MS from three independent biological replicates for each growth condition. Values are the average for the three replicates, and the significance of fold change was analyzed by ANOVA with P < 0.05. ( C ) Reverse-phase HPLC quantification of E. coli ATCC 25922 metabolites when fed aerobically with [ 14 C-methyl]−5′-methylthioadenosine. Unk, unknown (identified as 2-methylsulfinylethanol by LC-MS/MS, m / z = 109.0319), R T = 6.7 min; Met, methionine, R T = 8.0 min; MTR, methylthioribose, R T = 16.7 min; MT-EtOH, 2-methylthioethanol, R T = 21.7 min; IS, internal standard, R T = 24.0 min; and MTA, 5′-methylthioadenosine, R T = 31.8 min.
    Figure Legend Snippet: Sulfur from MTA cannot be salvaged by E. coli for growth. ( A ) ATCC 25922 maximum growth achieved after 24 hours measured by optical density at 600 nm when cultured with 1 mM of the indicated sulfur compound as the sole sulfur source. No further growth was observed after 24 hours. Average and standard deviation error bars are for n = 3 independent replicates. ( B ) Fold difference in the abundance of DHAP shunt-associated metabolites when ATCC 25922 was grown aerobically in the presence of 1 mM sulfate and 1 mM MTA versus grown aerobically in the presence of 1 mM sulfate only. Metabolites were resolved by LC-MS/MS from three independent biological replicates for each growth condition. Values are the average for the three replicates, and the significance of fold change was analyzed by ANOVA with P < 0.05. ( C ) Reverse-phase HPLC quantification of E. coli ATCC 25922 metabolites when fed aerobically with [ 14 C-methyl]−5′-methylthioadenosine. Unk, unknown (identified as 2-methylsulfinylethanol by LC-MS/MS, m / z = 109.0319), R T = 6.7 min; Met, methionine, R T = 8.0 min; MTR, methylthioribose, R T = 16.7 min; MT-EtOH, 2-methylthioethanol, R T = 21.7 min; IS, internal standard, R T = 24.0 min; and MTA, 5′-methylthioadenosine, R T = 31.8 min.

    Techniques Used: Cell Culture, Standard Deviation, Liquid Chromatography with Mass Spectroscopy

    methionine salvage pathways  (ATCC)


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    Structured Review

    ATCC methionine salvage pathways
    Salvage of S-adenosyl- l <t>-methionine</t> (SAM) utilization byproducts, 5-deoxy-pentose sugars, and 6-deoxy-hexose sugars in E. coli . ( A ) S -adenosyl- l -homocysteine (SAH), produced by methyltransferases, is recycled by the methionine cycle (a.k.a. active methyl cycle) ( , ). ( B and C ) The E. coli variation of the DHAP shunt. ( B ) For adenine salvage and detoxification of 5′-methylthioadenosine (MTA) and 5′-deoxyadenosine (5dAdo), all E. coli possess the multifunctional SAH/MTA/5dAdo nucleosidase (Pfs; a.k.a. MtnN) . ( C ) ExPEC strains possess the Pfs nucleosidase ( B ) and remainder of the DHAP shunt genes, which together compose a dual-purpose pathway for the conversion of 5′-deoxy-nucleosides in the form of MTA and 5dAdo or corresponding 5-deoxy-pentoses in the form of 5-deoxy- d -ribose and 5-methylthioribose, respectively, into the central carbon metabolite DHAP and acetaldehyde or (2-methylthio)acetaldehyde . ( D ) E. coli analogously metabolize 6-deoxy-hexose sugars in the form of l -fucose and l -rhamnose to DHAP and l -lactaldehyde. During anaerobic growth, l -lactaldehyde is primarily reduced to ( S )-1,2-propanediol as a terminal product, whereas during aerobic growth, it is primarily oxidized to l -lactate for carbon assimilation as pyruvate. Enzymes: Pfs (MtnN), SAH/MTA/5dAdo nucleosidase, E.C. 3.2.2.9; LuxS, S -ribosyl- l -homocysteine lyase, E.C. 4.4.1.21; MetH, methionine synthase, E.C. 2.1.1.13; MetE, methionine synthase, E.C. 2.1.1.4; MtnK, 5-methylthioribose/5-deoxyribose kinase, E.C. 2.7.1.100; MtnA, 5-methylthioribose-1-phosphate/5-deoxyribose-1-phosphate isomerase, E.C. 5.3.1.23; Ald2, 5-methylthioribulose-1-phosphate/5-deoxyribulose-1-phosphate aldolase, E.C. 4.1.2.62; ADH, alcohol dehydrogenase, E.C. 1.1.1.1 ; FucI, l -fucose isomerase, E.C. 5.3.1.25; FucK, l -fuculose kinase, E.C. 2.7.1.51; FucA, l -fuculose-1-phosphate aldolase, E.C. 4.1.2.17; RhaA, l -rhamnose isomerase, E.C. 5.3.1.14; RhaB, l -rhamnulose kinase, E.C. 2.7.1.5; RhaD, l -rhamnulose-1-phosphate aldolase, E.C. 4.1.2.19; FucO, ( S )−1,2 -propanediol oxidoreductase, E.C. 1.1.1.77; and AldA, l -lactaldehyde dehydrogenase, E.C. 1.2.1.22.
    Methionine Salvage Pathways, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Images

    1) Product Images from "Escherichia coli possessing the dihydroxyacetone phosphate shunt utilize 5′-deoxynucleosides for growth"

    Article Title: Escherichia coli possessing the dihydroxyacetone phosphate shunt utilize 5′-deoxynucleosides for growth

    Journal: Microbiology Spectrum

    doi: 10.1128/spectrum.03086-23

    Salvage of S-adenosyl- l -methionine (SAM) utilization byproducts, 5-deoxy-pentose sugars, and 6-deoxy-hexose sugars in E. coli . ( A ) S -adenosyl- l -homocysteine (SAH), produced by methyltransferases, is recycled by the methionine cycle (a.k.a. active methyl cycle) ( , ). ( B and C ) The E. coli variation of the DHAP shunt. ( B ) For adenine salvage and detoxification of 5′-methylthioadenosine (MTA) and 5′-deoxyadenosine (5dAdo), all E. coli possess the multifunctional SAH/MTA/5dAdo nucleosidase (Pfs; a.k.a. MtnN) . ( C ) ExPEC strains possess the Pfs nucleosidase ( B ) and remainder of the DHAP shunt genes, which together compose a dual-purpose pathway for the conversion of 5′-deoxy-nucleosides in the form of MTA and 5dAdo or corresponding 5-deoxy-pentoses in the form of 5-deoxy- d -ribose and 5-methylthioribose, respectively, into the central carbon metabolite DHAP and acetaldehyde or (2-methylthio)acetaldehyde . ( D ) E. coli analogously metabolize 6-deoxy-hexose sugars in the form of l -fucose and l -rhamnose to DHAP and l -lactaldehyde. During anaerobic growth, l -lactaldehyde is primarily reduced to ( S )-1,2-propanediol as a terminal product, whereas during aerobic growth, it is primarily oxidized to l -lactate for carbon assimilation as pyruvate. Enzymes: Pfs (MtnN), SAH/MTA/5dAdo nucleosidase, E.C. 3.2.2.9; LuxS, S -ribosyl- l -homocysteine lyase, E.C. 4.4.1.21; MetH, methionine synthase, E.C. 2.1.1.13; MetE, methionine synthase, E.C. 2.1.1.4; MtnK, 5-methylthioribose/5-deoxyribose kinase, E.C. 2.7.1.100; MtnA, 5-methylthioribose-1-phosphate/5-deoxyribose-1-phosphate isomerase, E.C. 5.3.1.23; Ald2, 5-methylthioribulose-1-phosphate/5-deoxyribulose-1-phosphate aldolase, E.C. 4.1.2.62; ADH, alcohol dehydrogenase, E.C. 1.1.1.1 ; FucI, l -fucose isomerase, E.C. 5.3.1.25; FucK, l -fuculose kinase, E.C. 2.7.1.51; FucA, l -fuculose-1-phosphate aldolase, E.C. 4.1.2.17; RhaA, l -rhamnose isomerase, E.C. 5.3.1.14; RhaB, l -rhamnulose kinase, E.C. 2.7.1.5; RhaD, l -rhamnulose-1-phosphate aldolase, E.C. 4.1.2.19; FucO, ( S )−1,2 -propanediol oxidoreductase, E.C. 1.1.1.77; and AldA, l -lactaldehyde dehydrogenase, E.C. 1.2.1.22.
    Figure Legend Snippet: Salvage of S-adenosyl- l -methionine (SAM) utilization byproducts, 5-deoxy-pentose sugars, and 6-deoxy-hexose sugars in E. coli . ( A ) S -adenosyl- l -homocysteine (SAH), produced by methyltransferases, is recycled by the methionine cycle (a.k.a. active methyl cycle) ( , ). ( B and C ) The E. coli variation of the DHAP shunt. ( B ) For adenine salvage and detoxification of 5′-methylthioadenosine (MTA) and 5′-deoxyadenosine (5dAdo), all E. coli possess the multifunctional SAH/MTA/5dAdo nucleosidase (Pfs; a.k.a. MtnN) . ( C ) ExPEC strains possess the Pfs nucleosidase ( B ) and remainder of the DHAP shunt genes, which together compose a dual-purpose pathway for the conversion of 5′-deoxy-nucleosides in the form of MTA and 5dAdo or corresponding 5-deoxy-pentoses in the form of 5-deoxy- d -ribose and 5-methylthioribose, respectively, into the central carbon metabolite DHAP and acetaldehyde or (2-methylthio)acetaldehyde . ( D ) E. coli analogously metabolize 6-deoxy-hexose sugars in the form of l -fucose and l -rhamnose to DHAP and l -lactaldehyde. During anaerobic growth, l -lactaldehyde is primarily reduced to ( S )-1,2-propanediol as a terminal product, whereas during aerobic growth, it is primarily oxidized to l -lactate for carbon assimilation as pyruvate. Enzymes: Pfs (MtnN), SAH/MTA/5dAdo nucleosidase, E.C. 3.2.2.9; LuxS, S -ribosyl- l -homocysteine lyase, E.C. 4.4.1.21; MetH, methionine synthase, E.C. 2.1.1.13; MetE, methionine synthase, E.C. 2.1.1.4; MtnK, 5-methylthioribose/5-deoxyribose kinase, E.C. 2.7.1.100; MtnA, 5-methylthioribose-1-phosphate/5-deoxyribose-1-phosphate isomerase, E.C. 5.3.1.23; Ald2, 5-methylthioribulose-1-phosphate/5-deoxyribulose-1-phosphate aldolase, E.C. 4.1.2.62; ADH, alcohol dehydrogenase, E.C. 1.1.1.1 ; FucI, l -fucose isomerase, E.C. 5.3.1.25; FucK, l -fuculose kinase, E.C. 2.7.1.51; FucA, l -fuculose-1-phosphate aldolase, E.C. 4.1.2.17; RhaA, l -rhamnose isomerase, E.C. 5.3.1.14; RhaB, l -rhamnulose kinase, E.C. 2.7.1.5; RhaD, l -rhamnulose-1-phosphate aldolase, E.C. 4.1.2.19; FucO, ( S )−1,2 -propanediol oxidoreductase, E.C. 1.1.1.77; and AldA, l -lactaldehyde dehydrogenase, E.C. 1.2.1.22.

    Techniques Used: Produced

    Sulfur from MTA cannot be salvaged by E. coli for growth. ( A ) ATCC 25922 maximum growth achieved after 24 hours measured by optical density at 600 nm when cultured with 1 mM of the indicated sulfur compound as the sole sulfur source. No further growth was observed after 24 hours. Average and standard deviation error bars are for n = 3 independent replicates. ( B ) Fold difference in the abundance of DHAP shunt-associated metabolites when ATCC 25922 was grown aerobically in the presence of 1 mM sulfate and 1 mM MTA versus grown aerobically in the presence of 1 mM sulfate only. Metabolites were resolved by LC-MS/MS from three independent biological replicates for each growth condition. Values are the average for the three replicates, and the significance of fold change was analyzed by ANOVA with P < 0.05. ( C ) Reverse-phase HPLC quantification of E. coli ATCC 25922 metabolites when fed aerobically with [ 14 C-methyl]−5′-methylthioadenosine. Unk, unknown (identified as 2-methylsulfinylethanol by LC-MS/MS, m / z = 109.0319), R T = 6.7 min; Met, methionine, R T = 8.0 min; MTR, methylthioribose, R T = 16.7 min; MT-EtOH, 2-methylthioethanol, R T = 21.7 min; IS, internal standard, R T = 24.0 min; and MTA, 5′-methylthioadenosine, R T = 31.8 min.
    Figure Legend Snippet: Sulfur from MTA cannot be salvaged by E. coli for growth. ( A ) ATCC 25922 maximum growth achieved after 24 hours measured by optical density at 600 nm when cultured with 1 mM of the indicated sulfur compound as the sole sulfur source. No further growth was observed after 24 hours. Average and standard deviation error bars are for n = 3 independent replicates. ( B ) Fold difference in the abundance of DHAP shunt-associated metabolites when ATCC 25922 was grown aerobically in the presence of 1 mM sulfate and 1 mM MTA versus grown aerobically in the presence of 1 mM sulfate only. Metabolites were resolved by LC-MS/MS from three independent biological replicates for each growth condition. Values are the average for the three replicates, and the significance of fold change was analyzed by ANOVA with P < 0.05. ( C ) Reverse-phase HPLC quantification of E. coli ATCC 25922 metabolites when fed aerobically with [ 14 C-methyl]−5′-methylthioadenosine. Unk, unknown (identified as 2-methylsulfinylethanol by LC-MS/MS, m / z = 109.0319), R T = 6.7 min; Met, methionine, R T = 8.0 min; MTR, methylthioribose, R T = 16.7 min; MT-EtOH, 2-methylthioethanol, R T = 21.7 min; IS, internal standard, R T = 24.0 min; and MTA, 5′-methylthioadenosine, R T = 31.8 min.

    Techniques Used: Cell Culture, Standard Deviation, Liquid Chromatography with Mass Spectroscopy

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    ATCC acid 1 65 methionine
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    C. amycolatum WT, C. amycolatum cob-, C. glucuronolyticum, C. glutamicum, and C. kefirresidentii were cultured in minimal cobamide-free medium with 0, 50, or 250 nM CoCl 2 concentrations. Intracellular and extracellular cobamides were estimated using an E. coli microbiological indicator assay . Cobamide vs. <t>methionine</t> presence in the samples was confirmed using both the E. coli metE- and E. coli metE - Δ metH indicator strains. Intracellular cobamides were quantified per gram of wet cell weight, and extracellular (CFSM) cobamides were quantified per mL of media. Intracellular cobamides were converted to their cyano form before testing in the indicator assay, and extracellular cobamides were tested in their native form.
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    C. amycolatum WT, C. amycolatum cob-, C. glucuronolyticum, C. glutamicum, and C. kefirresidentii were cultured in minimal cobamide-free medium with 0, 50, or 250 nM CoCl 2 concentrations. Intracellular and extracellular cobamides were estimated using an E. coli microbiological indicator assay . Cobamide vs. <t>methionine</t> presence in the samples was confirmed using both the E. coli metE- and E. coli metE - Δ <t>metH</t> indicator strains. Intracellular cobamides were quantified per gram of wet cell weight, and extracellular (CFSM) cobamides were quantified per mL of media. Intracellular cobamides were converted to their cyano form before testing in the indicator assay, and extracellular cobamides were tested in their native form.
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    C. amycolatum WT, C. amycolatum cob-, C. glucuronolyticum, C. glutamicum, and C. kefirresidentii were cultured in minimal cobamide-free medium with 0, 50, or 250 nM CoCl 2 concentrations. Intracellular and extracellular cobamides were estimated using an E. coli microbiological indicator assay . Cobamide vs. <t>methionine</t> presence in the samples was confirmed using both the E. coli metE- and E. coli metE - Δ <t>metH</t> indicator strains. Intracellular cobamides were quantified per gram of wet cell weight, and extracellular (CFSM) cobamides were quantified per mL of media. Intracellular cobamides were converted to their cyano form before testing in the indicator assay, and extracellular cobamides were tested in their native form.
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    C. amycolatum WT, C. amycolatum cob-, C. glucuronolyticum, C. glutamicum, and C. kefirresidentii were cultured in minimal cobamide-free medium with 0, 50, or 250 nM CoCl 2 concentrations. Intracellular and extracellular cobamides were estimated using an E. coli microbiological indicator assay . Cobamide vs. <t>methionine</t> presence in the samples was confirmed using both the E. coli metE- and E. coli metE - Δ <t>metH</t> indicator strains. Intracellular cobamides were quantified per gram of wet cell weight, and extracellular (CFSM) cobamides were quantified per mL of media. Intracellular cobamides were converted to their cyano form before testing in the indicator assay, and extracellular cobamides were tested in their native form.
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    C. amycolatum WT, C. amycolatum cob-, C. glucuronolyticum, C. glutamicum, and C. kefirresidentii were cultured in minimal cobamide-free medium with 0, 50, or 250 nM CoCl 2 concentrations. Intracellular and extracellular cobamides were estimated using an E. coli microbiological indicator assay . Cobamide vs. <t>methionine</t> presence in the samples was confirmed using both the E. coli metE- and E. coli metE - Δ <t>metH</t> indicator strains. Intracellular cobamides were quantified per gram of wet cell weight, and extracellular (CFSM) cobamides were quantified per mL of media. Intracellular cobamides were converted to their cyano form before testing in the indicator assay, and extracellular cobamides were tested in their native form.
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    C. amycolatum WT, C. amycolatum cob-, C. glucuronolyticum, C. glutamicum, and C. kefirresidentii were cultured in minimal cobamide-free medium with 0, 50, or 250 nM CoCl 2 concentrations. Intracellular and extracellular cobamides were estimated using an E. coli microbiological indicator assay . Cobamide vs. methionine presence in the samples was confirmed using both the E. coli metE- and E. coli metE - Δ metH indicator strains. Intracellular cobamides were quantified per gram of wet cell weight, and extracellular (CFSM) cobamides were quantified per mL of media. Intracellular cobamides were converted to their cyano form before testing in the indicator assay, and extracellular cobamides were tested in their native form.

    Journal: bioRxiv

    Article Title: Skin-associated Corynebacterium amycolatum shares cobamides

    doi: 10.1101/2024.04.28.591522

    Figure Lengend Snippet: C. amycolatum WT, C. amycolatum cob-, C. glucuronolyticum, C. glutamicum, and C. kefirresidentii were cultured in minimal cobamide-free medium with 0, 50, or 250 nM CoCl 2 concentrations. Intracellular and extracellular cobamides were estimated using an E. coli microbiological indicator assay . Cobamide vs. methionine presence in the samples was confirmed using both the E. coli metE- and E. coli metE - Δ metH indicator strains. Intracellular cobamides were quantified per gram of wet cell weight, and extracellular (CFSM) cobamides were quantified per mL of media. Intracellular cobamides were converted to their cyano form before testing in the indicator assay, and extracellular cobamides were tested in their native form.

    Article Snippet: To evaluate cobamide sharing on solid medium, a solid medium with no cobamides or methionine was prepared to support growth of both C. amycolatum, E. coli ATCC 14169 (metE - ), and E. coli metE - Δ metH , termed modified CM9.

    Techniques: Cell Culture

    (A) E. coli metE - (blue) was cultured with 0, 1, and 2.5 ng/mL cyanocobalamin standards (CnCbl), and (B) E. coli metE - Δ metH (pink) was cultured with 0, 10, and 100 μg/mL methionine to validate expected growth response of each strain, quantified in CFU/mL. Growth of E. coli metE - is proportional to the concentration of cobamides or methionine in the medium, and growth of E. coli metE - Δ metH is proportional to the concentration of only methionine, thus distinguishing between cobamides and methionine in supporting E. coli growth. C. amycolatum WT (WT) was co-cultured in minimal medium with (C) E. coli metE - (blue) or (D) E. coli metE - Δ metH (pink) under low (0 nM) or high (250 nM) cobalt concentrations. C. amycolatum cob - (cob-) was co-cultured in minimal medium with (E) E. coli metE - (blue) or (F) E. coli metE - Δ metH (pink) under low (0 nM) or high (250 nM) cobalt concentrations. (G) C. amycolatum CFU/mL was measured to demonstrate growth across each strain condition. n=9 replicates for each condition (3 technical replicates across 3 biological replicates). Solid lines represent the measured inoculum for E. coli metE - , E. coli metE - Δ metH , or C. amycolatum averaged across all replicates. FS = filter-sterilized C. amycolatum cell suspension, L = lysed C. amycolatum cell suspension.

    Journal: bioRxiv

    Article Title: Skin-associated Corynebacterium amycolatum shares cobamides

    doi: 10.1101/2024.04.28.591522

    Figure Lengend Snippet: (A) E. coli metE - (blue) was cultured with 0, 1, and 2.5 ng/mL cyanocobalamin standards (CnCbl), and (B) E. coli metE - Δ metH (pink) was cultured with 0, 10, and 100 μg/mL methionine to validate expected growth response of each strain, quantified in CFU/mL. Growth of E. coli metE - is proportional to the concentration of cobamides or methionine in the medium, and growth of E. coli metE - Δ metH is proportional to the concentration of only methionine, thus distinguishing between cobamides and methionine in supporting E. coli growth. C. amycolatum WT (WT) was co-cultured in minimal medium with (C) E. coli metE - (blue) or (D) E. coli metE - Δ metH (pink) under low (0 nM) or high (250 nM) cobalt concentrations. C. amycolatum cob - (cob-) was co-cultured in minimal medium with (E) E. coli metE - (blue) or (F) E. coli metE - Δ metH (pink) under low (0 nM) or high (250 nM) cobalt concentrations. (G) C. amycolatum CFU/mL was measured to demonstrate growth across each strain condition. n=9 replicates for each condition (3 technical replicates across 3 biological replicates). Solid lines represent the measured inoculum for E. coli metE - , E. coli metE - Δ metH , or C. amycolatum averaged across all replicates. FS = filter-sterilized C. amycolatum cell suspension, L = lysed C. amycolatum cell suspension.

    Article Snippet: To evaluate cobamide sharing on solid medium, a solid medium with no cobamides or methionine was prepared to support growth of both C. amycolatum, E. coli ATCC 14169 (metE - ), and E. coli metE - Δ metH , termed modified CM9.

    Techniques: Cell Culture, Concentration Assay, Suspension

    C. amycolatum WT or cob - cell suspension or controls (CnCbl, methionine, filtered cell suspension (FS), lysed cell suspension (L) were added to half of a minimal medium plate and incubated for 3 days, after which E. coli metE - (blue) or E. coli metE - Δ metH (pink) was struck out on the adjacent side of the plate. E. coli growth was recorded after 24 h. Growth summary for all conditions is indicated in (A). - indicates no growth and +, ++, +++ indicate increasing levels of growth, respectively. C. amycolatum WT lawn with (B) E. coli metE - and (C) E. coli metE - Δ metH. C. amycolatum cob - lawn with (D) E. coli metE - , (E) E. coli metE - + 100 ng/mL CnCbl, and (F) E. coli metE - Δ metH. E. coli metE - with (G) C. amycolatum filter-sterilized cell suspension and (H) C. amycolatum lysed cell suspension. Blank controls for (I) E. coli metE - and (J) E. coli metE - Δ metH . (K) E. coli metE - and (L) E. coli metE - Δ metH growth response to CnCbl and methionine standards.

    Journal: bioRxiv

    Article Title: Skin-associated Corynebacterium amycolatum shares cobamides

    doi: 10.1101/2024.04.28.591522

    Figure Lengend Snippet: C. amycolatum WT or cob - cell suspension or controls (CnCbl, methionine, filtered cell suspension (FS), lysed cell suspension (L) were added to half of a minimal medium plate and incubated for 3 days, after which E. coli metE - (blue) or E. coli metE - Δ metH (pink) was struck out on the adjacent side of the plate. E. coli growth was recorded after 24 h. Growth summary for all conditions is indicated in (A). - indicates no growth and +, ++, +++ indicate increasing levels of growth, respectively. C. amycolatum WT lawn with (B) E. coli metE - and (C) E. coli metE - Δ metH. C. amycolatum cob - lawn with (D) E. coli metE - , (E) E. coli metE - + 100 ng/mL CnCbl, and (F) E. coli metE - Δ metH. E. coli metE - with (G) C. amycolatum filter-sterilized cell suspension and (H) C. amycolatum lysed cell suspension. Blank controls for (I) E. coli metE - and (J) E. coli metE - Δ metH . (K) E. coli metE - and (L) E. coli metE - Δ metH growth response to CnCbl and methionine standards.

    Article Snippet: To evaluate cobamide sharing on solid medium, a solid medium with no cobamides or methionine was prepared to support growth of both C. amycolatum, E. coli ATCC 14169 (metE - ), and E. coli metE - Δ metH , termed modified CM9.

    Techniques: Suspension, Incubation

    C. amycolatum WT, C. amycolatum cob-, C. glucuronolyticum, C. glutamicum, and C. kefirresidentii were cultured in minimal cobamide-free medium with 0, 50, or 250 nM CoCl 2 concentrations. Intracellular and extracellular cobamides were estimated using an E. coli microbiological indicator assay . Cobamide vs. methionine presence in the samples was confirmed using both the E. coli metE- and E. coli metE - Δ metH indicator strains. Intracellular cobamides were quantified per gram of wet cell weight, and extracellular (CFSM) cobamides were quantified per mL of media. Intracellular cobamides were converted to their cyano form before testing in the indicator assay, and extracellular cobamides were tested in their native form.

    Journal: bioRxiv

    Article Title: Skin-associated Corynebacterium amycolatum shares cobamides

    doi: 10.1101/2024.04.28.591522

    Figure Lengend Snippet: C. amycolatum WT, C. amycolatum cob-, C. glucuronolyticum, C. glutamicum, and C. kefirresidentii were cultured in minimal cobamide-free medium with 0, 50, or 250 nM CoCl 2 concentrations. Intracellular and extracellular cobamides were estimated using an E. coli microbiological indicator assay . Cobamide vs. methionine presence in the samples was confirmed using both the E. coli metE- and E. coli metE - Δ metH indicator strains. Intracellular cobamides were quantified per gram of wet cell weight, and extracellular (CFSM) cobamides were quantified per mL of media. Intracellular cobamides were converted to their cyano form before testing in the indicator assay, and extracellular cobamides were tested in their native form.

    Article Snippet: The kanamycin resistance cassette was amplified from pKD4 (Supplemental Table 1) using primers with homology extensions that flank the upstream and downstream regions of methionine synthase metH in E. coli ATCC 14169 (Supplemental Table 2, homology extensions underlined).

    Techniques: Cell Culture

    (A) E. coli metE - (blue) was cultured with 0, 1, and 2.5 ng/mL cyanocobalamin standards (CnCbl), and (B) E. coli metE - Δ metH (pink) was cultured with 0, 10, and 100 μg/mL methionine to validate expected growth response of each strain, quantified in CFU/mL. Growth of E. coli metE - is proportional to the concentration of cobamides or methionine in the medium, and growth of E. coli metE - Δ metH is proportional to the concentration of only methionine, thus distinguishing between cobamides and methionine in supporting E. coli growth. C. amycolatum WT (WT) was co-cultured in minimal medium with (C) E. coli metE - (blue) or (D) E. coli metE - Δ metH (pink) under low (0 nM) or high (250 nM) cobalt concentrations. C. amycolatum cob - (cob-) was co-cultured in minimal medium with (E) E. coli metE - (blue) or (F) E. coli metE - Δ metH (pink) under low (0 nM) or high (250 nM) cobalt concentrations. (G) C. amycolatum CFU/mL was measured to demonstrate growth across each strain condition. n=9 replicates for each condition (3 technical replicates across 3 biological replicates). Solid lines represent the measured inoculum for E. coli metE - , E. coli metE - Δ metH , or C. amycolatum averaged across all replicates. FS = filter-sterilized C. amycolatum cell suspension, L = lysed C. amycolatum cell suspension.

    Journal: bioRxiv

    Article Title: Skin-associated Corynebacterium amycolatum shares cobamides

    doi: 10.1101/2024.04.28.591522

    Figure Lengend Snippet: (A) E. coli metE - (blue) was cultured with 0, 1, and 2.5 ng/mL cyanocobalamin standards (CnCbl), and (B) E. coli metE - Δ metH (pink) was cultured with 0, 10, and 100 μg/mL methionine to validate expected growth response of each strain, quantified in CFU/mL. Growth of E. coli metE - is proportional to the concentration of cobamides or methionine in the medium, and growth of E. coli metE - Δ metH is proportional to the concentration of only methionine, thus distinguishing between cobamides and methionine in supporting E. coli growth. C. amycolatum WT (WT) was co-cultured in minimal medium with (C) E. coli metE - (blue) or (D) E. coli metE - Δ metH (pink) under low (0 nM) or high (250 nM) cobalt concentrations. C. amycolatum cob - (cob-) was co-cultured in minimal medium with (E) E. coli metE - (blue) or (F) E. coli metE - Δ metH (pink) under low (0 nM) or high (250 nM) cobalt concentrations. (G) C. amycolatum CFU/mL was measured to demonstrate growth across each strain condition. n=9 replicates for each condition (3 technical replicates across 3 biological replicates). Solid lines represent the measured inoculum for E. coli metE - , E. coli metE - Δ metH , or C. amycolatum averaged across all replicates. FS = filter-sterilized C. amycolatum cell suspension, L = lysed C. amycolatum cell suspension.

    Article Snippet: The kanamycin resistance cassette was amplified from pKD4 (Supplemental Table 1) using primers with homology extensions that flank the upstream and downstream regions of methionine synthase metH in E. coli ATCC 14169 (Supplemental Table 2, homology extensions underlined).

    Techniques: Cell Culture, Concentration Assay, Suspension

    C. amycolatum WT or cob - cell suspension or controls (CnCbl, methionine, filtered cell suspension (FS), lysed cell suspension (L) were added to half of a minimal medium plate and incubated for 3 days, after which E. coli metE - (blue) or E. coli metE - Δ metH (pink) was struck out on the adjacent side of the plate. E. coli growth was recorded after 24 h. Growth summary for all conditions is indicated in (A). - indicates no growth and +, ++, +++ indicate increasing levels of growth, respectively. C. amycolatum WT lawn with (B) E. coli metE - and (C) E. coli metE - Δ metH. C. amycolatum cob - lawn with (D) E. coli metE - , (E) E. coli metE - + 100 ng/mL CnCbl, and (F) E. coli metE - Δ metH. E. coli metE - with (G) C. amycolatum filter-sterilized cell suspension and (H) C. amycolatum lysed cell suspension. Blank controls for (I) E. coli metE - and (J) E. coli metE - Δ metH . (K) E. coli metE - and (L) E. coli metE - Δ metH growth response to CnCbl and methionine standards.

    Journal: bioRxiv

    Article Title: Skin-associated Corynebacterium amycolatum shares cobamides

    doi: 10.1101/2024.04.28.591522

    Figure Lengend Snippet: C. amycolatum WT or cob - cell suspension or controls (CnCbl, methionine, filtered cell suspension (FS), lysed cell suspension (L) were added to half of a minimal medium plate and incubated for 3 days, after which E. coli metE - (blue) or E. coli metE - Δ metH (pink) was struck out on the adjacent side of the plate. E. coli growth was recorded after 24 h. Growth summary for all conditions is indicated in (A). - indicates no growth and +, ++, +++ indicate increasing levels of growth, respectively. C. amycolatum WT lawn with (B) E. coli metE - and (C) E. coli metE - Δ metH. C. amycolatum cob - lawn with (D) E. coli metE - , (E) E. coli metE - + 100 ng/mL CnCbl, and (F) E. coli metE - Δ metH. E. coli metE - with (G) C. amycolatum filter-sterilized cell suspension and (H) C. amycolatum lysed cell suspension. Blank controls for (I) E. coli metE - and (J) E. coli metE - Δ metH . (K) E. coli metE - and (L) E. coli metE - Δ metH growth response to CnCbl and methionine standards.

    Article Snippet: The kanamycin resistance cassette was amplified from pKD4 (Supplemental Table 1) using primers with homology extensions that flank the upstream and downstream regions of methionine synthase metH in E. coli ATCC 14169 (Supplemental Table 2, homology extensions underlined).

    Techniques: Suspension, Incubation