Description

Project Abstract

 This year, we are excited to introduce our innovative project, CoPlat, a modularized platform with adhesive properties that can immobilize functional proteins. In response to the existing limitations in related techniques, we have developed CoPlat to overcome these challenges, offering a novel and advantageous solution. This presentation aims to outline our project, highlighting CoPlat’s five important features using the acronym "POWER" and delineating our comprehensive plan for CoPlat's development in three crucial stages:


Stage 1 - Constructing CoPlat:

 In this phase, we use CsgA to link with barnacle proteins or mussel proteins, creating the foundation for CoPlat's construction.

Figure 1.The photo is of E. coli producing CoPlat and CoPlat anchor on the membrane.


Stage 2 - Expansion and diversification of CoPlat:
 We enhance CoPlat by employing a classifier to identify potentially adhesive proteins, broadening its applicability and utility.

Stage 3 - Ensuring the ability of modularization:  To demonstrate the modularity of CoPlat, we integrate GFP (Green Fluorescent Protein) onto the platform, showcasing its versatile functionality.

Figure 2.The photo is of E. coli producing CoPlat with functional proteins and anchor on the membrane.


 Ultimately, we successfully generated CoPlat, verified the property of modularization, and successfully optimized CoPlat’s performance through potential adhesive proteins.

Inspiration

 This year, our team, NYCU-Formosa, tries to focus on three issues: Heavy Metal Water Pollution, Formaldehyde Release in Manufactured Wood, and Cost of Immobilization. The following are the major drawbacks of these three issues.

(1) Immobilization

 Immobilization is an efficient technique and is widely used in industrial enzymatic reactions. However, current immobilization methods have limitations and drawbacks. For example, adsorption and embedding may physically immobilize in a non-fixed direction, which may block the enzyme's binding site and make it less efficient. Covalent bonding and cross-linking methods may cause chemical contamination during the manufacturing process. In addition, they are more costly than the direct use of enzymes.[1]

(2) Heavy metal water pollution

 Heavy metal water pollution is an urgent global issue that requires immediate attention and resolution. This is also a severe issue in Hsinchu Science Park, Taiwan, because many important semiconductors are located here.[2] If there is a lack of improper management of the wastewater, it will pose environmental and health risks. At present, Hsinchu Science Park uses electrolysis for such heavy metal ion-containing wastewater management. However, it has the disadvantage that this method cannot be used if the concentration of the metal ions is below the standard.[3]

(3) The formaldehyde in manufactured wood

 The manufactured wood is often soaked in chemicals and glued together during manufacturing, causing formaldehyde to form in the wood and then evaporate into the air during manufacturing and decoration.[4] The formaldehyde causes health problems when inhaled by humans.[5] The current solution is to use solid wood or low-formaldehyde lumber, but both are expensive and prone to insect infestation and mold.


In fact, there are already some solutions to these issues. However, some obstacles and limitations still need to be considered. Our team came up with a novel idea that can simply break the limitations: CoPlat.

When using CoPlat, it can easily immobilize the enzymes to use in the industry. In addition, it can also integrate metal ions binding proteins to absorb the metal ions specifically and increase the concentration of metal ions for further electrolysis. Moreover, it can not only be regarded as a simple adhesive to glue wood to reduce the release of formaldehyde but also be integrated with insect-repellent proteins to protect the wood from pests and mildew.

So, what is CoPlat? And what is its composition?

What is "CoPlat"?

CoPlat is a modularized platform, and its composition is the CsgA linking with adhesive proteins and anchoring to the membrane of bacteria. The platform's modularity stems from its flexible linker, which integrates various functional proteins. Enabled by adhesive proteins, CoPlat exhibits remarkable adhesion capabilities, allowing it to adhere to nearly any surface. To sum up, CoPlat is a modularized platform characterized by its adhesive properties, which not only enhance protein reaction rates but also expand the versatility and utility of E. coli in protein expression. We concluded five key characteristics of CoPlat:
(1) Predict adhesion
(2) Object-adherable
(3) Wide-ranging utility
(4) Easy to use
(5) Reinforcement
These highlights can make a "POWER" CoPlat.

Predict adhesion

 Over the past decade, the number of protein sequences in databases like UniProt and Pfam has grown rapidly. However, there has been a widening gap between the number of protein function annotations and the total number of sequences.[6] Consequently, machine learning has recently been increasingly employed to predict protein function classes.
 Our team extensively gathered information on adhesive proteins from UniProt[7], a protein database. We analyzed their characteristics and constructed a machine learning model. This approach allows us to accurately predict proteins with adhesive properties, expanding the pool of proteins usable with CoPlat.

Object-adherable

 CoPlat can adhere to surfaces and incorporates adhesive proteins, including Mussel Foot Proteins (MFPs) and Barnacle Cement Proteins (BCPs), etc. These adhesive proteins allow CoPlat to have the ability of adhesion, expanding its applicability to a broader array of surfaces.

Wide-ranging utility

 Benefiting from the modular design, our team envisions CoPlat serving a multitude of purposes. On the macroscopic scale, CoPlat finds utility in tasks such as heavy metal wastewater treatment, wood adhesive, and enzyme immobilization.
 Users have the capability to customize CoPlat's functional components to align with their specific needs. This adaptability empowers CoPlat to accurately fulfill the requirements of diverse applications and offers an array of possibilities for customization and integration.[8]

Easy to use

 Unlike complex industrial designs, utilizing CoPlat is a more streamlined process. You begin by designing the biobrick and then utilize the linker to integrate the functional proteins with CoPlat. Our products are biodegradable and sustainable, minimizing environmental impact and waste disposal, producing fewer harmful by-products, and consuming fewer non-renewable resources.[9]

Reinforcement

The amyloid protein CsgA, a primary cell membrane component in E. coli, is present in the extracellular matrix.[10] CsgA serves as an anchor, immobilizing functional proteins on the cell surface.[11] Additionally, its high surface area enhances the rate of protein reactions. In our project, we utilize CsgA to link adhesive proteins, thereby augmenting E. coli's adhesion capability to various surfaces.[12] Coplat also improves protein efficiency in applications through the property of immobilization.[13]

Conclusion

 To achieve the “POWER” CoPlat, we designed three stages to produce CoPlat and confirm the functions of it. More details in the Design part.


Reference
  1. Ycel S, Terziolu P and zime D (2012) Lipase Applications in Biodiesel Production. Biodiesel - Feedstocks, Production and Applications. InTech. DOI: 10.5772/52662.
  2. Hall, M. (2022, December 12). GLOBAL CHIP INDUSTRY PROJECTED TO INVEST MORE THAN $500 BILLION IN NEW FACTORY CONSTRUCTION STARTS BY 2024, SEMI REPORTS.
  3. Sheng Easy Technology Co., Ltd. Comprehensive Compilation of Heavy Metal Wastewater Treatment Techniques.
  4. Eco-Friendly Phenol–Urea–Formaldehyde Co-condensed Resin Adhesives Accelerated by Resorcinol for Plywood Manufacturing Bo Pang, Ming-Kan Li, Sheng Yang, Tong-Qi Yuan, Guan-Ben Du, and Run-Cang Sun ACS Omega 2018 3 (8), 8521-8528 DOI: 10.1021/acsomega.8b01286
  5. CHEN YI HUNG. (2002). The Effect to Wood Based Panels Workers' Lung Function When Exposing to Formaldehyde (National Defense Medical Center, Taipei, Taiwan (R.O.C.)).
  6. Bonetta, R., & Valentino, G. (2020). Machine learning techniques for protein function prediction. Proteins: Structure, Function, and Bioinformatics, 88(3), 397-413.
  7. UniProt: the universal protein knowledgebase. Nucleic acids research, 2017, 45.D1: D158-D169.
  8. DiMarco, R. L., & Heilshorn, S. C. (2012). Multifunctional materials through modular protein engineering. Advanced Materials, 24(29), 3923-3940.
  9. Zhang, Y. H. P., Sun, J., & Ma, Y. (2017). Biomanufacturing: history and perspective. Journal of Industrial Microbiology and Biotechnology, 44(4-5), 773-784.
  10. Tang, T. C., An, B., Huang, Y., Vasikaran, S., Wang, Y., Jiang, X., ... & Zhong, C. (2021). Materials design by synthetic biology. Nature Reviews Materials, 6(4), 332-350.
  11. Nguyen, P. Q., Botyanszki, Z., Tay, P. K. R., & Joshi, N. S. (2014). Programmable biofilm-based materials from engineered curli nanofibres. Nature communications, 5(1), 4945.
  12. Botyanszki, Z., Tay, P. K. R., Nguyen, P. Q., Nussbaumer, M. G., & Joshi, N. S. (2015). Engineered catalytic biofilms: Site‐specific enzyme immobilization onto E. coli curli nanofibers. Biotechnology and bioengineering, 112(10), 2016-2024.
  13. Garcia-Galan, C., Berenguer‐Murcia, Á., Fernandez‐Lafuente, R., & Rodrigues, R. C. (2011). Potential of different enzyme immobilization strategies to improve enzyme performance. Advanced Synthesis & Catalysis, 353(16), 2885-2904.