EcN Folding Capacity for DAF and Lactoferrin (comp-043)¶

Can an exact DAF or lactoferrin construct reach its native, functional state through an EcN expression and secretion route?

COMP-043 does not answer that question. Its disulfide-capacity bands, connectivity weights, protease scores, glycan assumptions, and composite outputs were uncalibrated. The numerical ordering, viability crossover, chassis comparison, and experiment priority are invalid. The COMP-043 tombstone is non-runnable; Git retains the retired implementation and outputs.

Evidence boundary¶

DAF and lactoferrin are disulfide-containing proteins with distinct native folds (In Vitro structural and annotation records; UniProt P08174 and P02788). That makes native-fold attainment a necessary measurement for each exact EcN construct. This page does not reproduce historical feature counts; reverify any count against the current primary record before using it as a design input.

The prior scan did not identify a calibrated DsbA/DsbC capacity rule that converts disulfide architecture into expression success for these exact secreted configurations (Mechanistic Extrapolation). Cytoplasmic oxidizing-strain precedents do not establish the capacity of the proposed periplasmic route.

Research conjecture — folding-system capacity may become a configuration-specific bottleneck

Grounded premises: DAF and lactoferrin require specific disulfide-connected native folds (In Vitro; UniProt P08174, UniProt P02788, and PMID 10089347). EcN route-specific expression, native-fold attainment, secretion, stability, and retained function have not been measured for the proposed constructs.

Novel leap: One or more exact EcN payload configurations may exceed the useful capacity of the baseline folding route, and DsbC co-expression may rescue some—not necessarily all—of that failure. No direct evidence establishes this for the proposed constructs.

Why it matters: A configuration-level result can redirect construct, secretion route, folding support, or chassis without declaring the payload class viable or infeasible.

Discriminating observation: Compare baseline and DsbC-co-expression arms for each exact construct. Measure expression, secretion, native disulfide connectivity or validated native-fold proxy, aggregation, route-relevant stability, and retained function. Advance only the configuration that meets prespecified functional and quality gates.

The result does not choose EcN or koji. A negative result kills only the tested construct × route × folding-support configuration.