Overview
- External ontology
- TOGO:M258 — Methanomicrobium Mobile Medium
- Applications
-
- Microbial cultivation
- Notes
- Source: https://togomedium.org/medium/M258 Original source: JCM - JCM_M266 Original URL: https://www.jcm.riken.jp/cgi-bin/jcm/jcm_grmd?GRMD=266 Superseded historical provenance (not a current formulation claim): Commercial Product: LB Medium (Luria-Bertani, Miller formulation) Supplier: Multiple suppliers (Difco, Sigma, etc.) Catalog: Multiple (L3522 Sigma, 244520 BD) Source: https://www.laboratorynotes.com/preparation-of-luria-bertani-lb-miller-broth/ Accessed: 2026-03-28 Constituent composition researched from product specification. The earlier LB commercial-product expansion was incorrect for M258/JCM266: three unrelated LB constituents are removed and alpha-methylbutyric acid is restored in the fatty-acid stock at the source batch amount, 0.5 g. The historical supplier paragraph above is retained only as superseded provenance. No chemical grounding is assigned to the restored ingredient. Original post-inoculation instruction: Pressurize the inoculated bottles to 200 kPa H2-CO2 (80:20, v/v).
Composition
flowchart LR
CultureMech_009157["methanomicrobium_mobile_medium"]
ing0_Distilled_water["Distilled water"]
CultureMech_009157 --> ing0_Distilled_water
CHEBI_15377["CHEBI:15377"]
ing0_Distilled_water -.-> CHEBI_15377
ing1_MgSO4_7H2O["MgSO4・7H2O"]
CultureMech_009157 --> ing1_MgSO4_7H2O
CHEBI_31795["CHEBI:31795"]
ing1_MgSO4_7H2O -.-> CHEBI_31795
ing2_NaCl["NaCl"]
CultureMech_009157 --> ing2_NaCl
CHEBI_26710["CHEBI:26710"]
ing2_NaCl -.-> CHEBI_26710
ing3_CaCl2_2H2O["CaCl2・2H2O"]
CultureMech_009157 --> ing3_CaCl2_2H2O
CHEBI_86158["CHEBI:86158"]
ing3_CaCl2_2H2O -.-> CHEBI_86158
ing4_KH2PO4["KH2PO4"]
CultureMech_009157 --> ing4_KH2PO4
CHEBI_63036["CHEBI:63036"]
ing4_KH2PO4 -.-> CHEBI_63036
ing5_K2HPO4["K2HPO4"]
CultureMech_009157 --> ing5_K2HPO4
CHEBI_131527["CHEBI:131527"]
ing5_K2HPO4 -.-> CHEBI_131527
ing6_Resazurin["Resazurin"]
CultureMech_009157 --> ing6_Resazurin
CHEBI_8806["CHEBI:8806"]
ing6_Resazurin -.-> CHEBI_8806
ing7_FeSO4_7H2O["FeSO4・7H2O"]
CultureMech_009157 --> ing7_FeSO4_7H2O
CHEBI_75836["CHEBI:75836"]
ing7_FeSO4_7H2O -.-> CHEBI_75836
ing8_NaHCO3["NaHCO3"]
CultureMech_009157 --> ing8_NaHCO3
CHEBI_32139["CHEBI:32139"]
ing8_NaHCO3 -.-> CHEBI_32139
ing9__NH4_2SO4["(NH4)2SO4"]
CultureMech_009157 --> ing9__NH4_2SO4
CHEBI_62946["CHEBI:62946"]
ing9__NH4_2SO4 -.-> CHEBI_62946
ing10_Yeast_extract__BD_Difco_["Yeast extract (BD-Difco)"]
CultureMech_009157 --> ing10_Yeast_extract__BD_Difco_
ing11_Trypticase_peptone__BD_BBL_["Trypticase peptone (BD-BBL)"]
CultureMech_009157 --> ing11_Trypticase_peptone__BD_BBL_
ing12_Carbon_dioxide_gas["Carbon dioxide gas"]
CultureMech_009157 --> ing12_Carbon_dioxide_gas
CHEBI_16526["CHEBI:16526"]
ing12_Carbon_dioxide_gas -.-> CHEBI_16526
ing13_N2["N2"]
CultureMech_009157 --> ing13_N2
CHEBI_17997["CHEBI:17997"]
ing13_N2 -.-> CHEBI_17997
ing14_Hydrogen_gas["Hydrogen gas"]
CultureMech_009157 --> ing14_Hydrogen_gas
CHEBI_18276["CHEBI:18276"]
ing14_Hydrogen_gas -.-> CHEBI_18276
Ingredients (15)
| # | Ingredient | Concentration | Ontology | Notes |
|---|---|---|---|---|
| 1 |
Distilled water
H2O 18.015 g/mol
|
0.66 L |
CHEBI:15377
water
|
Role: Solvating media; Properties: Inorganic compound, Defined component, Simple component [Merged 2 duplicates: 660.0, 20.0] Source water batch input: 660 ml = 0.66 L. This is not a measured final volume. The earlier duplicate-summing note is superseded: it combined separate preparation scopes and source units. This node carries only the batch input at the cited source row; other occurrences are separate scoped ingredients. |
| 2 |
MgSO4・7H2O
7H2O.Mg.O4S 246.473 g/mol
|
CHEBI:31795
magnesium sulfate heptahydrate
|
Role: Mineral source; Properties: Inorganic compound, Defined component, Simple component Source batch input: 0.13 g. Final preparation volume is not independently established; the unsupported g/L assertion is withdrawn, not replaced by an assumed one-liter concentration. | |
| 3 |
NaCl
Cl.Na 58.443 g/mol
|
CHEBI:26710
sodium chloride
|
Role: Mineral source; Properties: Inorganic compound, Defined component, Simple component Source batch input: 0.6 g. Final preparation volume is not independently established; the unsupported g/L assertion is withdrawn, not replaced by an assumed one-liter concentration. | |
| 4 |
CaCl2・2H2O
Ca.2Cl.2H2O 147.014 g/mol
|
CHEBI:86158
calcium chloride dihydrate
|
Role: Mineral source; Properties: Inorganic compound, Defined component, Simple component Source batch input: 8 mg. Final preparation volume is not independently established; the unsupported g/L assertion is withdrawn, not replaced by an assumed one-liter concentration. | |
| 5 |
KH2PO4
H2O4P.K 136.084 g/mol
|
CHEBI:63036
potassium dihydrogen phosphate
|
Role: Buffer; Properties: Inorganic compound, Defined component, Simple component Source batch input: 0.3 g. Final preparation volume is not independently established; the unsupported g/L assertion is withdrawn, not replaced by an assumed one-liter concentration. | |
| 6 |
K2HPO4
HO4P.2K 174.174 g/mol
|
CHEBI:131527
dipotassium hydrogen phosphate
|
Role: Buffer; Properties: Inorganic compound, Defined component, Simple component Source batch input: 0.3 g. Final preparation volume is not independently established; the unsupported g/L assertion is withdrawn, not replaced by an assumed one-liter concentration. | |
| 7 |
Resazurin
C12H7NO4 229.191 g/mol
|
CHEBI:8806
Resazurin
|
Role: pH dependent redox indicator; Properties: Organic compound, Defined component, Simple component Source batch input: 1 mg. Final preparation volume is not independently established; the unsupported g/L assertion is withdrawn, not replaced by an assumed one-liter concentration. | |
| 8 |
FeSO4・7H2O
Fe.7H2O.O4S 278.013 g/mol
|
CHEBI:75836
iron(2+) sulfate heptahydrate
|
Role: Mineral source; Properties: Inorganic compound, Defined component, Simple component Source batch input: 2 mg. Final preparation volume is not independently established; the unsupported g/L assertion is withdrawn, not replaced by an assumed one-liter concentration. | |
| 9 |
NaHCO3
CHO3.Na 84.006 g/mol
|
CHEBI:32139
sodium hydrogencarbonate
|
Role: Buffer; Properties: Inorganic compound, Defined component, Simple component Source batch input: 2 g. Final preparation volume is not independently established; the unsupported g/L assertion is withdrawn, not replaced by an assumed one-liter concentration. | |
| 10 |
(NH4)2SO4
2H4N.O4S 132.141 g/mol
|
CHEBI:62946
ammonium sulfate
|
Role: Nitrogen source; Properties: Inorganic compound, Defined component, Simple component Source batch input: 0.3 g. Final preparation volume is not independently established; the unsupported g/L assertion is withdrawn, not replaced by an assumed one-liter concentration. | |
| 11 | Yeast extract (BD-Difco) | Properties: Undefined component Source batch input: 1 g. Final preparation volume is not independently established; the unsupported g/L assertion is withdrawn, not replaced by an assumed one-liter concentration. | ||
| 12 | Trypticase peptone (BD-BBL) | Properties: Undefined component Source batch input: 1 g. Final preparation volume is not independently established; the unsupported g/L assertion is withdrawn, not replaced by an assumed one-liter concentration. | ||
| 13 |
Carbon dioxide gas
CO2 44.009 g/mol
|
CHEBI:16526
carbon dioxide
|
Properties: Gas Source atmosphere context, not a fixed gas input or dissolved concentration. Removed the unsupported VARIABLE placeholder. | |
| 14 |
N2
N2 28.014 g/mol
|
CHEBI:17997
dinitrogen
|
Properties: Gas Source atmosphere context, not a fixed gas input or dissolved concentration. Removed the unsupported VARIABLE placeholder. | |
| 15 |
Hydrogen gas
H2 2.016 g/mol
|
CHEBI:18276
dihydrogen
|
Properties: Gas Source atmosphere context, not a fixed gas input or dissolved concentration. Removed the unsupported VARIABLE placeholder. |
Preparation
- Mix ingredients, except NaHCO3, L-cysteine·HCl·H2O and Na2S·9H2O, and adjust pH to 6.5.
- Bring to a boil for 5-10 sec, then, cool down under a stream of H2-CO2 (80:20, v/v) and add NaHCO3 to the medium.
- Dispense the medium into suitable culture vessels (e.g., 20 ml of the medium in 120 ml serum bottles) under a stream of H2-CO2 (80:20, v/v) and seal with butyl rubber stoppers.
- After autoclaving, stand the medium overnight.
- Separately autoclave cysteine·HCl and Na2S·9H2O as 5% solutions, respectively, under a N2 atmosphere.
- Prior to inoculation, aseptically and anaerobically add the L-cysteine·HCl·H2O and Na2S·9H2O solutions.
Solutions
Rumen fluid, clarified (see below)
Dose: 0.3 L
| Ingredient | Concentration | Notes |
|---|---|---|
| clarified rumen fluid | Role: Nutrient source; Properties: Undefined component Unquantified clarified-fluid product description, not an additional independent dose. The enclosing stock descriptor alone holds the 300 ml receiving-batch input. Removed the unsupported VARIABLE placeholder. |
Source stock addition: 300 ml = 0.3 L per receiving batch, not grams per liter or a dose per final liter.
Expand only this named preparation, not the complete referenced medium. Water input does not establish final stock volume; no final-medium rate is inferred.
Trace minerals (see Medium [M142])
Dose: 0.01 L
| Ingredient | Concentration | Notes |
|---|---|---|
| Distilled water | 1 L | Source water batch input: 1 L = 1 L. This is not a measured final volume. |
| MgSO4・7H2O | Source batch input: 3 g. Final preparation volume is not independently established; the source batch amount is retained without asserting a final concentration. | |
| NaCl | Source batch input: 1 g. Final preparation volume is not independently established; the source batch amount is retained without asserting a final concentration. | |
| CaCl2・2H2O | Source batch input: 0.1 g. Final preparation volume is not independently established; the source batch amount is retained without asserting a final concentration. | |
| Na2MoO4・2H2O | Source batch input: 0.01 g. Final preparation volume is not independently established; the source batch amount is retained without asserting a final concentration. | |
| H3BO3 | Source batch input: 0.01 g. Final preparation volume is not independently established; the source batch amount is retained without asserting a final concentration. | |
| FeSO4・7H2O | Source batch input: 0.1 g. Final preparation volume is not independently established; the source batch amount is retained without asserting a final concentration. | |
| ZnSO4・7H2O | Source batch input: 0.1 g. Final preparation volume is not independently established; the source batch amount is retained without asserting a final concentration. | |
| CuSO4・5H2O | Source batch input: 0.01 g. Final preparation volume is not independently established; the source batch amount is retained without asserting a final concentration. | |
| CoSO4・7H2O | Source batch input: 0.1 g. Final preparation volume is not independently established; the source batch amount is retained without asserting a final concentration. | |
| Nitrilotriacetic acid | Source batch input: 1.5 g. Final preparation volume is not independently established; the source batch amount is retained without asserting a final concentration. | |
| MnSO4・xH2O | Source batch input: 0.5 g. Final preparation volume is not independently established; the source batch amount is retained without asserting a final concentration. | |
| AlK(SO4)2 | Source batch input: 0.01 g. Final preparation volume is not independently established; the source batch amount is retained without asserting a final concentration. |
Cross-reference to Medium M142
Source stock addition: 10 ml = 0.01 L per receiving batch, not grams per liter or a dose per final liter.
Expand only this named preparation, not the complete referenced medium. Water input does not establish final stock volume; no final-medium rate is inferred.
KOH solution
Stock pH-adjustment reagent with no fixed dose or numeric strength.
Trace vitamins (see Medium [M190])
Dose: 0.01 L
| Ingredient | Concentration | Notes |
|---|---|---|
| Distilled water | 1 L | Source water batch input: 1 L = 1 L. This is not a measured final volume. |
| Biotin | Source batch input: 2 mg. Final preparation volume is not independently established; the source batch amount is retained without asserting a final concentration. | |
| p--Aminobenzoic acid | Source batch input: 5 mg. Final preparation volume is not independently established; the source batch amount is retained without asserting a final concentration. | |
| Thiamine・HCl | Source batch input: 5 mg. Final preparation volume is not independently established; the source batch amount is retained without asserting a final concentration. | |
| Calcium pantothenate | Source batch input: 5 mg. Final preparation volume is not independently established; the source batch amount is retained without asserting a final concentration. | |
| Pyridoxine・HCl | Source batch input: 10 mg. Final preparation volume is not independently established; the source batch amount is retained without asserting a final concentration. | |
| Folic acid | Source batch input: 2 mg. Final preparation volume is not independently established; the source batch amount is retained without asserting a final concentration. | |
| Vitamin B12 | Source batch input: 0.1 mg. Final preparation volume is not independently established; the source batch amount is retained without asserting a final concentration. | |
| Riboflavin | Source batch input: 5 mg. Final preparation volume is not independently established; the source batch amount is retained without asserting a final concentration. | |
| Nicotinic acid | Source batch input: 5 mg. Final preparation volume is not independently established; the source batch amount is retained without asserting a final concentration. | |
| Lipoic acid | Source batch input: 5 mg. Final preparation volume is not independently established; the source batch amount is retained without asserting a final concentration. |
Cross-reference to Medium M190
Source stock addition: 10 ml = 0.01 L per receiving batch, not grams per liter or a dose per final liter.
Expand only this named preparation, not the complete referenced medium. Water input does not establish final stock volume; no final-medium rate is inferred.
Fatty acid mixture (see below)
Dose: 0.02 L
| Ingredient | Concentration | Notes |
|---|---|---|
| Distilled water | 0.02 L | Role: Solvating media; Properties: Inorganic compound, Defined component, Simple component [Merged 2 duplicates: 660.0, 20.0] Source water batch input: 20 ml = 0.02 L. This is not a measured final volume. The earlier duplicate-summing note is superseded: it combined separate preparation scopes and source units. This node carries only the batch input at the cited source row; other occurrences are separate scoped ingredients. |
| Valeric acid | Role: Carbon source; Properties: Organic compound, Defined component, Simple component Source batch input: 0.5 g. Final preparation volume is not independently established; the unsupported g/L assertion is withdrawn, not replaced by an assumed one-liter concentration. | |
| Isovaleric acid | Role: Carbon source; Properties: Organic compound, Defined component, Simple component Source batch input: 0.5 g. Final preparation volume is not independently established; the unsupported g/L assertion is withdrawn, not replaced by an assumed one-liter concentration. | |
| Isobutyric acid | Role: Carbon source; Properties: Organic compound, Defined component, Simple component Source batch input: 0.5 g. Final preparation volume is not independently established; the unsupported g/L assertion is withdrawn, not replaced by an assumed one-liter concentration. | |
| α-Methylbutyric acid | Source batch input: 0.5 g. Final preparation volume is not independently established; the source batch amount is retained without asserting a final concentration. |
Source stock addition: 20 ml = 0.02 L per receiving batch, not grams per liter or a dose per final liter.
Expand only this named preparation, not the complete referenced medium. Water input does not establish final stock volume; no final-medium rate is inferred.
NaOH
Properties: Defined component, Inorganic compound, Solution
Concentrated NaOH adjusts only the fatty-acid stock to pH 7.5; no fixed dose or numeric strength is given. Removed the unsupported VARIABLE placeholder.
Na2S・9H2O separately prepared solution
| Ingredient | Concentration | Notes |
|---|---|---|
| Na2S・9H2O | Role: Reducing agent; Properties: Inorganic compound, Defined component, Simple component Source batch input: 0.5 g. Final preparation volume is not independently established; the unsupported g/L assertion is withdrawn, not replaced by an assumed one-liter concentration. This is the contribution specified for the receiving medium batch, not the mass for preparing an unspecified volume of stock. |
Separate reducing-agent preparation; composition notes retain the 0.5 g receiving-batch contribution, not a stock-batch recipe. Stock solvent volume and addition volume are unspecified. The source 5% label has unspecified percentage basis; no numeric stock strength or derived dose is asserted.
L--Cysteine・HCl・H2O separately prepared solution
| Ingredient | Concentration | Notes |
|---|---|---|
| L--Cysteine・HCl・H2O | Properties: Defined component, Simple component, Organic compound Source batch input: 0.5 g. Final preparation volume is not independently established; the unsupported g/L assertion is withdrawn, not replaced by an assumed one-liter concentration. This is the contribution specified for the receiving medium batch, not the mass for preparing an unspecified volume of stock. |
Separate reducing-agent preparation; composition notes retain the 0.5 g receiving-batch contribution, not a stock-batch recipe. Stock solvent volume and addition volume are unspecified. The source 5% label has unspecified percentage basis; no numeric stock strength or derived dose is asserted.
Target organisms
No target organisms are recorded.
Provenance
| Timestamp | Curator | Action | Notes |
|---|---|---|---|
| 2026-01-27T06:35:54.714451Z | togo-import | Imported from TOGO Medium | Source: JCM, ID: M258 |
| 2026-01-28T19:48:08.424156Z | chebi-enrichment | Added CHEBI ontology terms to ingredients | Enriched using MicrobeMediaParam and MediaDive chemical mappings |
| 2026-02-03T00:00:00+00:00 | schema-defaulter-v1.0 | Applied schema defaults and normalizations | Added default concentration for ingredient: Carbon dioxide gas; Added default concentration for ingredient: N2; Added default concentration for ingredient: Hydrogen gas; Added default concentration for ingredient: clarified rumen fluid; Added default concentration for ingredient: NaOH |
| 2026-03-09T08:04:13.602030+00:00 | solution-migrator-v1.0 | Migrated solutions from ingredients to solutions array | Moved 2 solution(s) from ingredients |
| 2026-03-13T03:08:40.591568Z | data-quality-cleanup-v1.0 | Data quality fixes | Merged 1 duplicate ingredient(s) |
| 2026-03-13T06:20:03.360367Z | culturemech-id-assigner-v1.0 | Assigned CultureMech ID | Assigned stable identifier: CultureMech:009157 |
| 2026-03-14T00:00:00+00:00 | schema-defaulter-v1.0 | Applied schema defaults and normalizations | Added default solution name; Added default solution name |
| 2026-03-17T06:29:42.000000Z | mediaingredientmech-enrichment-v1.0 | Added MediaIngredientMech links | Linked ingredients to MediaIngredientMech identifiers |
| 2026-03-17T19:46:22.085870+00:00 | normalize_media_names_to_snake_case.py | NAME_NORMALIZED | Normalized name from "Methanomicrobium Mobile Medium" to "methanomicrobium_mobile_medium" for consistent matching |
| 2026-05-16T09:03:14.403294+00:00 | migrate_legacy_fields.py | MIGRATED_LEGACY_FIELDS | curation_history.date->timestamp=2 |
| 2026-06-05T07:40:37.669560+00:00 | mim-legacy-to-chebi-migration-v1.0 | Refreshed legacy MediaIngredientMech links to CHEBI keying | Replaced 11 legacy mediaingredientmech_term link(s) with mediaingredientmech_chebi_term (id-safe: ingredient's own CHEBI + label). MIM deprecated the MediaIngredientMech:NNNNNN id scheme. |
| 2026-06-10T06:57:27.319387+00:00 | mgso4-heptahydrate-term-fix-v1.0 | Corrected MgSO4·7H2O primary term grounding to heptahydrate | Re-grounded 1 heptahydrate-labelled ingredient(s) from CHEBI:32599 (generic magnesium sulfate) to CHEBI:31795 (magnesium sulfate heptahydrate). Generic/hexahydrate labels untouched. |
| 2026-06-10T07:46:49.313497+00:00 | primary-term-label-canonicalization-v1.0 | Canonicalized primary term label to OBO canonical (OAK) | 3 primary term(s); ingredient name resolved by OAK sqlite:obo:chebi. |
| 2026-10-08T20:12:07.152932+00:00 | codex | REPAIRED_M258_IDENTITY_AND_M260_NESTED_STOCK_WITH_EVIDENCE | Corrected stock boundaries and unsupported quantity assertions with exact batch-amount evidence. M258 supersedes its erroneous LB expansion and restores alpha-methylbutyric acid, rumen/fatty-acid preparations and separate reducing agents. M260 nests the trace-mineral dose inside MJ(-N) seawater. Preserved unrelated annotations and append-only history. No final volume, percent basis or chemical grounding inferred. |