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SO3-BS3 Lithium-ion Battery Bioleaching Consortium

Community ID CommunityMech:000385
Ecological State ENGINEERED
Environment anaerobic brewing-wastewater NCM bioleaching culture — ontology label: laboratory environment
ENVO:01001405 · HabitatMech ↗

A two-member engineered Shewanella oneidensis MR-1 and Bacillus subtilis 168 synthetic consortium for metal oxide leaching from spent LiNixCoyMn1-x-yO2 lithium-ion battery cathode black mass using Moutai brewing wastewater as the organic leaching medium. The final SO3-BS3 design pairs S. oneidensis SO3, engineered for riboflavin synthesis and Deinococcus IrrE-mediated metal-ion stress resistance, with B. subtilis BS3, engineered for ethanol detoxification, riboflavin production, and IrrE-mediated resistance. In anaerobic bottle cultures with 0.2 g/L black mass, the SO3-BS3 consortium increased total dissolved Li, Ni, Co, and Mn to 102.4 mg/L by reducing NCM through MtrC/OmcA- and riboflavin-mediated extracellular electron transfer.

Taxonomy

Taxon Ontology ID Functional Roles Abundance
Shewanella oneidensis MR-1 SO3 — ontology label: Shewanella oneidensis NCBITaxon:70863 · TaxonMech · TraitMech
ELECTROGEN ELECTRON_DONOR
ABUNDANT
  • PMID:42733304 - SUPPORT (IN_VITRO)
    "SO3 SO1 with the Ptac:: ribADEHC operon from Bacillus subtilis 168 and irrE gene from Deinococcus radiodurans R1 integrated at pHG13"
Bacillus subtilis 168 BS3 — ontology label: Bacillus subtilis NCBITaxon:1423 · TaxonMech · TraitMech ABUNDANT
  • PMID:42733304 - SUPPORT (IN_VITRO)
    "BS3 BS2 with the P43:: irrE gene from Deinococcus radiodurans R1 integrated at pCE2"

Ecological Interactions

The graph shows only explicitly linked interaction participants. Community members without an asserted participant link are omitted; membership alone does not imply an interaction.

Ecological interaction network for SO3-BS3 Lithium-ion Battery Bioleaching Consortium Bipartite graph where circles represent taxa and non-circular symbols represent interactions. Rare or untyped interactions share the Other symbol; exact curated types remain in the interaction details.
Taxon
Other

BS3 ethanol detoxification of brewing wastewater

Bacillus subtilis BS3 converts residual ethanol in brewing wastewater to acetate via heterologous alcohol dehydrogenase and aldehyde dehydrogenase, removing an inhibitor of S. oneidensis extracellular electron transfer. This supports facilitation of SO3 electron transfer, but does not establish that BS3 is unaffected by SO3 as required for COMMENSALISM.

Source Taxon: Bacillus subtilis 168 BS3

Target Taxon: Shewanella oneidensis MR-1 SO3

Metabolites: ethanol (CHEBI:16236 · MediaIngredientMech · CMMMech), acetate (CHEBI:30089 · MediaIngredientMech · CMMMech)

Downstream Effects:
→ MtrC/OmcA and riboflavin NCM reduction

Ethanol removal relieves inhibition of Shewanella electron transfer and thereby supports NCM leaching in the authors' consortium model. The detoxification comparisons were conducted in predecessor strains; the final SO3-BS3 synthesis retains this function, without separately isolating its quantitative contribution from the other modifications.

Evidence

  • PMID:42733304 - SUPPORT (IN_VITRO)
    "strain BS3 in the SO3‐BS3 consortium eliminated the ethanol inhibition on EET of S. oneidensis by converting residual ethanol in the brewing wastewater into acetate via the Adh‐AldH dual enzyme cascade reaction."

MtrC/OmcA and riboflavin NCM reduction

Shewanella oneidensis SO3 catabolizes lactate from brewing wastewater and transfers electrons to NCM black mass either directly through outer-membrane cytochromes MtrC/OmcA or indirectly through riboflavin, reducing NCM metal oxides and releasing Li, Ni, Co, and Mn into solution. Final-consortium cytochrome perturbations and the reduced-riboflavin assay support the EET interpretation. The proposed direct and shuttle routes are not a quantitative partition of in-consortium electron flux.

Participating Taxa: Shewanella oneidensis MR-1 SO3, Bacillus subtilis 168 BS3

Metabolites: riboflavin (CHEBI:17015 · MediaIngredientMech · CMMMech), lithium(1+) (CHEBI:49713 · MediaIngredientMech · CMMMech), nickel(2+) (CHEBI:49786 · MediaIngredientMech · CMMMech), cobalt(2+) (CHEBI:48828 · MediaIngredientMech · CMMMech), manganese(2+) (CHEBI:29035 · MediaIngredientMech · CMMMech)

Evidence

  • PMID:42733304 - SUPPORT (IN_VITRO)
    "The generated electrons were then used to reduce NCM via the EET mechanism. Specifically, the electrons generated from strain SO3 could be either directly transferred to NCM via outer membrane c‐Cyts MtrC and OmcA, or first transferred to riboflavin molecules via MtrC and OmcA."
  • PMID:42733304 - SUPPORT (IN_VITRO)
    "strain BS3 in the SO3‐BS3 consortium eliminated the ethanol inhibition on EET of S. oneidensis by converting residual ethanol in the brewing wastewater into acetate via the Adh‐AldH dual enzyme cascade reaction. Also, strain BS3 synthesized riboflavin, which was employed as an electron shuttle for promoting the EET of S. oneidensis."
  • PMID:42733304 - SUPPORT (IN_VITRO)
    "confirming that the disruption of MtrC and OmcA significantly inhibited the EET process in strain SO3."

Knowledge gaps & discussions (2)

KNOWLEDGE_GAP OPEN The source established a 14-day laboratory proof of concept but explicitly left larger-scale, long-term stability unresolved, especially under natural variation in raw brewing-wastewater composition.

Raw brewing wastewater varied by batch and the authors identified large-scale reproducibility as a future optimization target, so the current record should not assert demonstrated industrial stability.

Attaches to: growth_media#Moutai brewing wastewater with spent NCM black mass

1 evidence item(s)
full_text SUPPORT PMID:42733304

the long‐term stability and reproducibility of the microbial consortium at a larger scale needed to be thoroughly considered, in particular under the condition of natural fluctuations in raw brewing wastewater compositions

Records the authors' stated scale-up uncertainty without asserting it as a demonstrated limitation.

KNOWLEDGE_GAP OPEN What is the reciprocal fitness effect and relative contribution of the retained processes?

Primary predecessor comparisons support ethanol removal, electron-transfer improvement and leaching. Final-consortium perturbations and riboflavin evidence support the retained reduction node. Neither benefit to SO3 nor greater leaching establishes neutral fitness of BS3 in the exact pair, so the optional COMMENSALISM type is removed without denying the supported facilitation. Direct-versus-shuttle flux and the isolated contribution of detoxification in the final consortium are not quantified by the cited comparisons. The existing scale-up discussion remains unchanged.

Attaches to: ecological_interactions#BS3 ethanol detoxification of brewing wastewater, ecological_interactions#MtrC/OmcA and riboflavin NCM reduction

1 evidence item(s)
IN_VITRO SUPPORT PMID:42733304

confirming that the ethanol inhibition of the EET of the SO1‐BS1 consortium was significantly alleviated

Identifies the predecessor consortium used for the detoxification comparison.

Environmental Factors

Factor Value Unit
Total leached Li, Ni, Co, and Mn 102.4 mg/L total dissolved metal ions after 14 days N/A
  • PMID:42733304 - SUPPORT (IN_VITRO)
    "Specifically, the total leached metal ions reached 102.4 mg L−1, which comprised 80.8, 8.5, 5.2, and 7.9 mg L−1 for Li, Ni, Co, and Mn, respectively"
Ni and Co cytotoxicity 6 mg/L Ni2+ and 3 mg/L Co2+ challenge N/A
  • PMID:42733304 - SUPPORT (IN_VITRO)
    "When exposed to 6 mg L−1 Ni2+ and 3 mg L−1 Co2+, the growth of strain SO2 was notably inhibited, with its maximum cell density decreasing from 1.941 (in the absence of Ni2+ and Co2+) to 1.367 (Figure 4e ). In comparison, strain SO3, which overexpressed IrrE, attained higher cell density (OD600, max = 2.291) under the identical conditions (Figure 4e )."
  • PMID:42733304 - SUPPORT (IN_VITRO)
    "Live‐dead staining also revealed that strains SO2 and BS2 exhibited more pronounced red fluorescence in the presence of Ni2+ and Co2+ (Figure 4g ), indicating a higher prevalence of dead cells due to Ni2+ and Co2+ exposure. The overexpression of IrrE in strains SO3 and BS3 resulted in a noticeable reduction in the abundance of dead cells (Figure 4g ), consistent with the observed growth patterns."

Growth Media

Chloramphenicol-supplemented Luria-Bertani pre-culture

Composition

Component Concentration Unit CHEBI
sodium chloride · MediaIngredientMech 10 g/L N/A
yeast extract · MediaIngredientMech 5 g/L N/A
peptone · MediaIngredientMech 10 g/L N/A
chloramphenicol · MediaIngredientMech 0.2 g/L N/A

Preparation notes: Each member was first grown separately overnight in LB with chloramphenicol for plasmid maintenance before transfer to the brewing-wastewater assay.

Evidence

PMID:42733304

"To establish a consortium of B. subtilis and S. oneidensis, both microorganisms were initially cultured in Luria‐Bertani (LB) medium containing 10 g L−1 NaCl, 5 g L−1 yeast extract, and 10 g L−1 peptone"

PMID:42733304

"supplemented with 0.2 g L−1 chloramphenicol for plasmid"

Moutai brewing wastewater with spent NCM black mass

Temperature 30 degrees Celsius
Atmosphere ANAEROBIC

Composition

Component Concentration Unit CHEBI
brewing wastewater · MediaIngredientMech N/A N/A N/A
LiNixCoyMn1-x-yO2 black mass · MediaIngredientMech 0.2 g/L N/A
isopropyl beta-D-thiogalactopyranoside · MediaIngredientMech 0.5 mM N/A

Preparation notes: SO3 and BS3 overnight cultures were transferred into Moutai brewing wastewater with IPTG for gene expression, then moved to anaerobic bottles with NCM black mass for leaching.

Evidence

PMID:42733304

"Overnight cultures of each strain were subsequently inoculated into a medium formulated with brewing water collected from a brewing factory in Moutai Town (Guizhou) at a 1% (v/v) ratio, induced by the isopropyl‐β‐d‐thiogalactoside (IPTG, 0.5 mm, Heowns, Tianjin) for gene expression. Following a brief period of subculturing, the culture was transferred into anaerobic bottles, supplemented with 0.2 g L−1 of black mass"

PMID:42733304

"Anaerobic fermentation was then conducted at 30°C for a period of 14 days to facilitate the dissolution of LIBs."

Cross-Mech Suite Links

Explore related entities across the CultureBotAI X-Mech knowledge bases:

HabitatMech: anaerobic brewing-wastewater NCM bioleaching culture ↗ TaxonMech: Shewanella oneidensis MR-1 SO3 ↗ TraitMech: Shewanella oneidensis MR-1 SO3 ↗ TaxonMech: Bacillus subtilis 168 BS3 ↗ TraitMech: Bacillus subtilis 168 BS3 ↗ CMMMech (Critical Minerals & REE) ↗ CultureMech Growth Media ↗ PathwayMech Pathways ↗