ATCC 2039 Broth (Twin Pack) l AS-1441
$125.00 AUD
Documentation
ATCC 2039 Broth (Twin Pack)
Official ATCC Medium 2039 | Acidithiobacillus ferrooxidans Culture Medium | Solution A + Solution B | Cat. No. AS-1441
Cat. No. AS-1441 Official ATCC 2039 Formula pH 2.3 ⚠ DO NOT AUTOCLAVE 🏢 Bioleaching Research 🏢 Acid Mine Drainage ✓ HiMedia M1963 Equivalent 🇦🇺 Made in Melbourne
Solution A — Salt Base
Inorganic Salts + Wolfe's Mineral
(NH₄)₂SO₄ + MgSO₄ + K₂HPO₄ + 5 mL Wolfe's Mineral in 800 mL | Adjust pH 2.3 with H₂SO₄ | Filter sterilise 0.22 µm
Solution B — Iron Source
FeSO₄·7H₂O — 20 g / 200 mL
Dissolve in 200 mL DI water | Filter sterilise IMMEDIATELY | Use within 1 hour | Never store solution
⚠️ DO NOT AUTOCLAVE: Autoclaving oxidises Fe²⁺ → Fe³⁺, precipitating iron hydroxide and destroying the medium. Filter sterilisation (0.22 µm) only for both solutions.
Cat. No.
AS-1441
Reference
ATCC 2039
Final pH
2.3 ± 0.1
Sterilisation
0.22 µm filter
Incubation
26–30°C + shaking
📄 Full DocumentationCOA, TDS & SDS every batch
🇦🇺 Australian StockNo import delays
⚡ Same-Week DispatchMelbourne warehouse
🔬 Technical SupportDirect from our team
Overview
ATCC 2039 Broth Twin Pack (AS-1441) is manufactured exactly to the official ATCC Medium 2039 specification (sourced directly from atcc.org) for the cultivation of Acidithiobacillus ferrooxidans ATCC 23270 and related acidophilic chemolithotrophs. The medium is supplied as a twin pack: Solution A (inorganic salts with Wolfe's Mineral Solution incorporated) + Solution B (ferrous sulphate heptahydrate, 20 g per 200 mL).
This is the only locally manufactured Australian source for ATCC Medium 2039, supplying the complete official formula including the full Wolfe's Mineral Solution trace element mix. No substitutions or simplifications.
Why a Twin Pack?
The ATCC official protocol specifies that Solution A and Solution B must be prepared and filter-sterilised separately before aseptic combination. This is because:
1. FeSO₄ stability: Fe²⁺ oxidises rapidly to Fe³⁺ in the presence of air, moisture, or elevated pH. Storing it with the salt base would cause oxidation before use, precipitating iron as Fe(OH)₃.
2. DO NOT AUTOCLAVE either solution: Autoclaving at 121°C oxidises Fe²⁺ → Fe³⁺ and precipitates iron hydroxide, destroying the medium's function as an energy source for Acidithiobacillus.
3. pH adjustment before filter sterilisation: Solution A must be adjusted to pH 2.3 with H₂SO₄ before filter sterilisation — not after.
1. FeSO₄ stability: Fe²⁺ oxidises rapidly to Fe³⁺ in the presence of air, moisture, or elevated pH. Storing it with the salt base would cause oxidation before use, precipitating iron as Fe(OH)₃.
2. DO NOT AUTOCLAVE either solution: Autoclaving at 121°C oxidises Fe²⁺ → Fe³⁺ and precipitates iron hydroxide, destroying the medium's function as an energy source for Acidithiobacillus.
3. pH adjustment before filter sterilisation: Solution A must be adjusted to pH 2.3 with H₂SO₄ before filter sterilisation — not after.
Wolfe's Mineral Solution — Why It Matters
The official ATCC 2039 formula includes 5 mL of Wolfe's Mineral Solution per litre. This is a complete trace element supplement containing 12 components including nitrilotriacetic acid (chelating agent), manganese, cobalt, zinc, copper, aluminium, boron, and molybdenum.
Acidithiobacillus ferrooxidans is an autotroph — it synthesises all organic molecules from CO₂. Without pre-formed amino acids or vitamins from peptone/yeast extract, it depends entirely on trace minerals supplied in the medium for enzyme cofactors, cell membrane function, and electron transport. Wolfe's Mineral Solution ensures these are present.
AS-1441 includes the complete Wolfe's Mineral Solution formula incorporated into Solution A — exactly as specified by ATCC.
Acidithiobacillus ferrooxidans is an autotroph — it synthesises all organic molecules from CO₂. Without pre-formed amino acids or vitamins from peptone/yeast extract, it depends entirely on trace minerals supplied in the medium for enzyme cofactors, cell membrane function, and electron transport. Wolfe's Mineral Solution ensures these are present.
AS-1441 includes the complete Wolfe's Mineral Solution formula incorporated into Solution A — exactly as specified by ATCC.
Applications
⛏
Bioleaching R&D
Cu, Au, U recovery from low-grade ores — heap leach process development
💧
Acid Mine Drainage
AMD characterisation, bioremediation, iron-oxidising bacteria isolation
🌎
Environmental
Acidophile isolation — mine sites, acid springs, volcanic environments
🧪
Mining Industry
Biohydrometallurgy R&D, mineral sulphide oxidation studies (Rio Tinto, BHP, etc.)
📊
Reference Strain
ATCC 23270 maintenance and propagation medium
📚
Research & Teaching
Extreme environment microbiology, chemolithotrophy demonstration
FAQs
❓ Why must pH be exactly 2.3?
At pH above 3.0, Fe³⁺ precipitates as iron hydroxide Fe(OH)₃ — the orange rust precipitate — and becomes unavailable as an energy source. At pH below 2.0, growth is inhibited. The ATCC protocol specifies pH 2.3 as the target. Verify with a calibrated pH meter before filter sterilisation. Use sterile dilute H₂SO₄ only — NOT HCl, which inhibits iron oxidation in this organism.
❓ A yellow precipitate forms after combining solutions — is this normal?
Yes — the ATCC official protocol states: "A yellow precipitate is normal; it becomes darker as the iron oxidizes." This is colloidal Fe³⁺ and is expected. The presence of yellow/orange colour during incubation is actually the growth indicator — it shows that A. ferrooxidans is actively oxidising Fe²⁺ to Fe³⁺ as its energy-yielding reaction.
❓ Why must Solution B be filter-sterilised immediately after dissolution?
FeSO₄ solution oxidises rapidly in air — Fe²⁺ becomes Fe³⁺ within hours, precipitating at pH above 2.5. Once Solution B is dissolved, it must be filter-sterilised within 30–60 minutes and combined immediately with sterile Solution A. The ATCC protocol states: "Stir Solution B to dissolve and quickly filter-sterilize." Never store prepared Solution B.
❓ Is AS-1441 equivalent to HiMedia M1963?
Yes — AS-1441 follows the same official ATCC Medium 2039 specification as HiMedia M1963, including the complete Wolfe's Mineral Solution. AS-1441 is manufactured in Melbourne with same-week dispatch and full Australian batch documentation — no import delays.
Cross-Reference / Equivalents
| Source | Product | Cat. No. |
|---|---|---|
| ATCC (official reference) | ATCC Medium 2039 — Acidithiobacillus ferrooxidans Medium | ATCC 2039 |
| HiMedia | Modified 9K Medium Base (Twin Pack) | M1963 |
| Silverman & Lundgren (1959) | Original 9K Medium formulation | — |
Product Specifications
| Product Name | ATCC 2039 Broth (Twin Pack) — Solution A + Solution B |
| Catalogue Number | AS-1441 |
| Reference Standard | Official ATCC Medium 2039 specification (atcc.org) | Silverman & Lundgren 9K Medium (1959) |
| Target Organism | Acidithiobacillus ferrooxidans ATCC 23270 and related acidophilic chemolithotrophs |
| Pack Format | Twin Pack — Solution A powder (salts + Wolfe's Mineral) + Solution B powder (FeSO₄·7H₂O) — packaged separately |
| Sterilisation | Filter sterilise ONLY (0.22 µm membrane) — DO NOT AUTOCLAVE either solution |
| Final pH | 2.3 ± 0.1 (adjust Solution A with sterile dilute H₂SO₄ — verify with calibrated pH meter) |
| Incubation | 26–30°C, aerobic, shaking 100–150 rpm, 4–14 days |
| Growth indicator | Yellow-orange colour (Fe³⁺ accumulation) — normal per ATCC protocol |
| Shelf life | 2 years from manufacture in original sealed packaging |
| Equivalents | HiMedia M1963 | ATCC Medium 2039 reference |
Solution A Formula — Per Litre of Final Medium (Official ATCC 2039)
| Ingredient | Amount | Function |
|---|---|---|
| Diammonium Sulphate ((NH₄)₂SO₄) | 0.8 g | Sole nitrogen source for autotrophic growth |
| Magnesium Sulphate Heptahydrate (MgSO₄·7H₂O) | 2.0 g | Essential macro-mineral; enzyme cofactor |
| Dipotassium Hydrogen Phosphate (K₂HPO₄) | 0.4 g | Phosphorus source; pH buffer |
| Wolfe's Mineral Solution (see below) | 5.0 mL | Complete trace element supplement per ATCC specification |
| DI / Purified Water | 800 mL | Adjust pH to 2.3 with H₂SO₄, then filter sterilise |
Wolfe's Mineral Solution — Complete Formula (Official ATCC 2039)
| Ingredient | g / L | Notes |
|---|---|---|
| Nitrilotriacetic acid | 1.500 | Chelating agent — add first to ~500 mL water, adjust pH 6.5 with KOH to dissolve |
| Magnesium Sulphate Heptahydrate (MgSO₄·7H₂O) | 3.000 | Macro-mineral supplement |
| Manganese Sulphate Monohydrate (MnSO₄·H₂O) | 0.500 | Trace mineral; enzyme cofactor |
| Sodium Chloride (NaCl) | 1.000 | Ionic balance |
| Ferrous Sulphate Heptahydrate (FeSO₄·7H₂O) | 0.100 | Trace iron |
| Cobalt Chloride Hexahydrate (CoCl₂·6H₂O) | 0.100 | Trace mineral; vitamin B12 synthesis |
| Calcium Chloride (CaCl₂) | 0.100 | Trace mineral; membrane function |
| Zinc Sulphate Heptahydrate (ZnSO₄·7H₂O) | 0.100 | Trace mineral; enzyme cofactor |
| Copper Sulphate Pentahydrate (CuSO₄·5H₂O) | 0.010 | Trace mineral; cytochrome oxidase |
| Aluminium Potassium Sulphate (AlK(SO₄)₂·12H₂O) | 0.010 | Trace mineral |
| Boric Acid (H₃BO₃) | 0.010 | Trace mineral |
| Sodium Molybdate Dihydrate (Na₂MoO₄·2H₂O) | 0.010 | Trace mineral; nitrogenase cofactor |
| DI / Purified Water | to 1000 mL | Bring to volume after adding all compounds one at a time |
ⓘ Wolfe's Mineral preparation note (from ATCC): Add nitrilotriacetic acid to approximately 500 mL of water and adjust pH to 6.5 with KOH to dissolve the compound. Bring volume to 1000 mL with remaining water and add remaining compounds one at a time.
Solution B Formula — Per Litre of Final Medium
| Ingredient | Amount | Function |
|---|---|---|
| Ferrous Sulphate Heptahydrate (FeSO₄·7H₂O) | 20.0 g | PRIMARY ENERGY SOURCE — A. ferrooxidans oxidises Fe²⁺ → Fe³⁺ as sole energy-yielding reaction. Do NOT reduce or substitute. |
| DI / Purified Water | 200 mL | Dissolve and filter-sterilise IMMEDIATELY. Do not store prepared solution. |
Preparation Protocol (Official ATCC 2039 Method)
⚠️ DO NOT AUTOCLAVE either Solution A or Solution B. Autoclaving oxidises Fe²⁺ → Fe³⁺ and precipitates iron hydroxide, making the medium unsuitable for iron-oxidising bacteria.
1
Prepare Wolfe's Mineral Solution: Add nitrilotriacetic acid to ~500 mL DI water, adjust pH to 6.5 with KOH to dissolve. Bring to 1000 mL and add remaining mineral compounds one at a time. Set aside 5.0 mL for Solution A.
2
Prepare Solution A: Dissolve (NH₄)₂SO₄ (0.8 g), MgSO₄·7H₂O (2.0 g), and K₂HPO₄ (0.4 g) in 800 mL DI water. Add 5.0 mL Wolfe's Mineral Solution. Mix well.
3
Adjust pH of Solution A to 2.3 with sterile dilute H₂SO₄. Verify with calibrated pH meter. Do NOT use HCl.
4
Filter sterilise Solution A through 0.22 µm membrane into a sterile vessel.
5
Prepare Solution B: Dissolve 20.0 g FeSO₄·7H₂O in 200 mL DI water. Stir to dissolve quickly.
6
Filter sterilise Solution B IMMEDIATELY through 0.22 µm membrane. Use within 30–60 minutes of dissolution.
7
Combine aseptically: Add sterile Solution B to sterile Solution A. Final volume: 1 litre. A yellow precipitate is normal — it becomes darker as iron oxidises (per ATCC protocol).
8
Dispense and inoculate. Incubate at 26–30°C aerobically with shaking (100–150 rpm) for 4–14 days.
Literature References
| # | Reference |
|---|---|
| 1 | ATCC. Medium 2039: Acidithiobacillus ferrooxidans Medium. American Type Culture Collection. atcc.org [Primary reference — formula sourced directly from official ATCC PDF] |
| 2 | Silverman MP, Lundgren DG. Studies on the chemoautotrophic iron bacterium Ferrobacillus ferrooxidans. J Bacteriol. 1959;77(5):642–647. [Original 9K medium] |
| 3 | Wolin EA, et al. Wolfe's Mineral Solution. J Bacteriol. 1963;81:911–918. |
| 4 | Kelly DP, Wood AP. Reclassification of some species of Thiobacillus. Int J Syst Evol Microbiol. 2000;50:511–516. |
| 5 | ISO 11133:2014. Microbiology of food and water — Performance testing of culture media. Geneva: ISO. |
📄 Full 16-section GHS SDS available (Australian WHS Regulations 2023 / GHS 7th Edition) — support@ausamics.com.au
Section 1 — Identification
| Product Name | ATCC 2039 Broth Twin Pack (AS-1441) — Solution A + Solution B |
| Intended Use | Culture medium for Acidithiobacillus ferrooxidans. Laboratory and research use only. |
| Supplier | AuSaMicS Pty Ltd | ABN 56 676 640 467 | 31 Longview CT, Thomastown VIC 3074 |
| Emergency | Poisons Information Centre: 13 11 26 (24 hr) |
| Phone | +61 412 520 598 | support@ausamics.com.au |
Section 2 — Hazard Identification
| Solution A (powder) | NOT classified as hazardous under Australian WHS Regulations 2023. |
| Solution B (FeSO₄·7H₂O powder) | NOT classified as hazardous at supplied concentrations. Mild irritant in bulk. |
| Prepared medium | CORROSIVE — pH 2.3. GHS05 pictogram. Signal Word: DANGER. H314: severe skin burns and eye damage. |
| H₂SO₄ (user-added) | CORROSIVE — handle with acid PPE. Not supplied by AuSaMicS. Consult H₂SO₄ SDS. |
| Precautionary (prepared) | P280: wear gloves, eye protection, face shield. P305+P351+P338: if in eyes — rinse 15+ min, seek emergency medical attention. P301+P330+P331: if ingested — do not induce vomiting, call 13 11 26. |
Key Components — Section 3
| Component | CAS | Solution | Hazard |
|---|---|---|---|
| (NH₄)₂SO₄ | 7783-20-2 | A | Not hazardous at use level |
| MgSO₄·7H₂O | 10034-99-8 | A + Wolfe's | Not hazardous at use level |
| K₂HPO₄ | 7758-11-4 | A | Mild irritant at high concentration |
| Nitrilotriacetic acid | 139-13-9 | Wolfe's | Irritant; possible reproductive toxicant at high doses — use PPE |
| FeSO₄·7H₂O | 7782-63-0 | B + Wolfe's | Irritant in powder; corrosive medium at pH 2.3 |
| H₂SO₄ (user-added) | 7664-93-9 | User adds to A | CORROSIVE — not supplied |
First Aid & PPE Summary
| Skin (powder) | Wash soap and water 15 min |
| Skin (prepared medium) | Flush with large amounts of water 20 min. Seek medical attention. |
| Eyes (prepared medium) | Immediately rinse 15+ min. Emergency medical attention. pH 2.3 is corrosive. |
| Ingestion | Do NOT induce vomiting. Call 13 11 26 immediately. |
| PPE — powder | Safety glasses, nitrile gloves, lab coat, P1 respirator for bulk weighing |
| PPE — prepared medium | Chemical splash goggles, acid-resistant gloves, lab coat — mandatory |
| Spill (prepared) | Neutralise with NaHCO₃. Prevent drain entry — pH 2.3 harmful to aquatic environment. |
| Disposal | Neutralise to pH 6–8. Iron hydroxide precipitate as inorganic waste per local EPA requirements. |
| Transport | Powder: not DG. Prepared acidic medium: verify corrosive liquid classification before transport. |
Quality Specifications
| Parameter | Specification | Typical Result | Method |
|---|---|---|---|
| Solution A appearance | White to off-white free-flowing powder | Conforms | Visual |
| Solution B appearance | Pale green to blue-green crystalline powder | Conforms | Visual |
| Solution B purity (FeSO₄·7H₂O) | ≥ 99.0% | 99.4% | Titration |
| Solution B iron content (as Fe) | 19.0 – 20.0% | 19.7% | AAS |
| Final pH (prepared, 25°C) | 2.3 ± 0.1 | 2.3 | pH meter (calibrated) |
| Solution A moisture | ≤ 3.0% | 1.4% | Loss on drying |
| Growth — A. ferrooxidans ATCC 23270 | Growth + yellow-orange colour by day 7–14 at 26–30°C | Growth confirmed | QC organism test |
| Sterility (uninoculated) | No growth at 30°C / 7 days | No growth | Sterility check |
| Batch COA | Lot-specific for both Solution A and Solution B | Included every order | — |
Pack Sizes
| 10L Twin Pack | Solution A: ~33 g | Solution B: 200 g | Makes 10 litres prepared medium |
| 25L Twin Pack ⭐ | Solution A: ~82 g | Solution B: 500 g | Makes 25 litres prepared medium — most popular |
| Custom | Contact support@ausamics.com.au for bulk or project-specific quantities |
Manufacturing & Documentation
Manufactured by AuSaMicS Pty Ltd
31 Longview CT, Thomastown VIC 3074, Australia | ABN 56 676 640 467
✓ Formulated exactly to official ATCC Medium 2039 specification (atcc.org)
✓ Complete Wolfe's Mineral Solution (12 components) incorporated in Solution A
✓ Solution B: FeSO₄·7H₂O ≥99.0% — lot-specific iron content on COA
✓ COA, TDS, SDS included with every order
✓ Australian stock — same-week dispatch, no import delays
✓ Only locally manufactured ATCC 2039 in Australia
31 Longview CT, Thomastown VIC 3074, Australia | ABN 56 676 640 467
✓ Formulated exactly to official ATCC Medium 2039 specification (atcc.org)
✓ Complete Wolfe's Mineral Solution (12 components) incorporated in Solution A
✓ Solution B: FeSO₄·7H₂O ≥99.0% — lot-specific iron content on COA
✓ COA, TDS, SDS included with every order
✓ Australian stock — same-week dispatch, no import delays
✓ Only locally manufactured ATCC 2039 in Australia
⚠️ For laboratory and research use only. Prepared medium is corrosive (pH 2.3). Acid-resistant gloves and chemical splash goggles mandatory. Not for food, therapeutic, veterinary, or household use.