Functional Proteins

We have contributed 14 new basic parts this year, including two novel engineered proteins—CBM3-HBP-CBM3 and CBM3-EV1-CBM3—designed as modular reinforcement modules for cellulose-based biomaterials. These are accompanied by two specialized Part Collections: one comprising a scaffold protein for multi-enzyme complex display on Saccharomyces cerevisiae, and another offering a selection of secretion signal peptides for Pichia pastoris expression. The CBM3-fused proteins were successfully expressed in E. coli with high solubility and integrated into straw-based fiber spinning systems. Notably, fibers incorporated with CBM3-EV1-CBM3 achieved a tensile strength of 2.56 cN/dtex, meeting industrial requirements for apparel textiles. These parts and collections provide future iGEM teams with functional, validated building blocks to enhance the mechanical properties and secretion efficiency of bio-based materials in a simple and programmable manner.

Part Number	Name	Type	Description
BBa_257L7ODW	CBM3-HBP1-CBM3	Coding	Encodes Histidine Rich Protein with carbohydrate-binding module CBM3 fused to both N- and C-termini.
BBa_25MUHCU8	CBM3-Ev1-CBM3	Coding	Encodes Eumeta variegata fibroin with carbohydrate-binding module CBM3 fused to both N- and C-termini.

Enzymes and Scaffold

Part Number	Name	Type	Description
BBa_250G1O1P	lcc1-RcdocI	Coding	Encodes a Laccase from Coriolopsis trogii, fused to dockerin from Ruminiclostridium cellulolyticum on C-terminus.
BBa_25NE2MXG	vpl2- RfdocI	Coding	Encodes a Versatile Peroxidase from Pleurotus eryngii, fused to dockerin from Ruminococcus flavefaciens on C-terminus.
BBa_25J82O1O	gh61-3-AtdocI	Coding	Encodes a Lytic Polysaccharide Monooxygenase from Neurospora crassa, fused to dockerin from Acetivibrio thermocellus on C-terminus.
BBa_25O180WL	LINKS-Scaffold	Coding	Encodes a scaffold protein incorporated by cohesin domains CipAcoh2, ScaBcoh4, and CipCcoh2. Fused to AGA2 of the a-agglutinin system for cell surface display.

The parts above are collected in our part collection (UUID：3cdf196b-d70d-4389-bc77-8fc680d8234b). This part collection offers a programmable platform for constructing scaffold-based enzyme complexes through orthogonal cohesin-dockerin interactions. This versatile system supports diverse applications, including the creation of enhanced enzymatic cascades for biosynthetic pathways and improved enzyme stability through scaffold-mediated immobilization. It provides standardized, reusable components that can be widely applied in biocatalysis and environmental applications.

Signal Peptides

Part Number	Name	Type	Description
BBa_25H90G8W/a>	MF4I	Coding	Encodes a resynthesized signal peptide originally derived from Aspergillus niger 113.
BBa_258L7IYG	α-factor ∆57–70	Coding	Encodes a truncated signal peptide originally derived from Saccharomyces cerevisiae. Primarily used in the Pichia pastoris expression system.
BBa_25NMRINZ	α-factor ∆57–70 Plus HL28	Coding	Encodes a modified version of the Saccharomyces cerevisiae α-factor signal peptide, able to enhance secretion of target proteins.
BBa_25ULQTOE	nSB	Coding	Encodes the CalB native signal peptide used for the expression of Candida antarctica lipase.
BBa_25UJBO2O	Dse4	Coding	Encodes a signal peptide derived from P. pastoris. Can drive heterologous protein secretion.
BBa_25KUDUIH	Msb2	Coding	Encodes a secretory-expression-driving signal peptide from mucin-family protein Msb2 native to Saccharomyces cerevisiae.
BBa_25NNTL4X	Gas1	Coding	Encodes the signal peptide of β-1,3-glucanosyltransferase Gas1 from Saccharomyces cerevisiae.
BBa_25K7XO3F	Gh61-3 SP	Coding	Encodes the endogenous signal peptide of the GH61-3 (LPMO) enzyme originating from Neurospora crassa.

The parts above are collected in our part collection (UUID：8ac5ca62-f2e4-4472-bb5b-06c0475b6985). This part collection provides eight experimentally validated secretion signals for Pichia pastoris, specifically designed to overcome the traditional bottleneck in recombinant protein expression. By offering a set of characterized signal peptides, this toolkit enables iGEM teams to bypass tedious screening processes and directly enhance protein secretion efficiency, significantly accelerating project development in biocatalysis and metabolic engineering.