Production and Characterization of Amelogenin-ELP Fusion Proteins
(2024) KBKM01 20241Pure and Applied Biochemistry
- Abstract
- This study aimed to engineer and characterize a series of Amelogenin-ELP fusion proteins to explore their potential as biomaterials with enhanced properties. Eleven constructs (176-186) were designed and expressed in E. coli BL21(DE3) cells, with successful purification confirmed by SDS-PAGE. Dynamic Light Scattering (DLS) revealed that constructs 181, 185, and 186 formed nanospheres similar to full-length amelogenin, while constructs 176 showed larger aggregates. Self-assembly experiments demonstrated that construct 183 could form nanoribbons under standard and acidic conditions, with calcium ions playing a crucial role. Hydrogel formation experiments indicated that construct 176 was more prone to gelation, especially in the presence of... (More)
- This study aimed to engineer and characterize a series of Amelogenin-ELP fusion proteins to explore their potential as biomaterials with enhanced properties. Eleven constructs (176-186) were designed and expressed in E. coli BL21(DE3) cells, with successful purification confirmed by SDS-PAGE. Dynamic Light Scattering (DLS) revealed that constructs 181, 185, and 186 formed nanospheres similar to full-length amelogenin, while constructs 176 showed larger aggregates. Self-assembly experiments demonstrated that construct 183 could form nanoribbons under standard and acidic conditions, with calcium ions playing a crucial role. Hydrogel formation experiments indicated that construct 176 was more prone to gelation, especially in the presence of CaCl₂ and KH₂PO₄, whereas construct 183 showed no gelation, highlighting the significance of engineered modifications. Film formation studies revealed distinct differences based on pH and additives, with additive-enhanced films exhibiting greater flexibility.
The ability of fusion proteins to form hydrogels and films was explored under varying pH and ionic conditions, revealing that specific constructs and conditions could be optimized to create biomaterials with desirable mechanical properties. The formation of nanoribbons and the role of additives in enhancing protein interactions were particularly noted for their potential in creating durable, flexible biomaterials. These findings suggest that Amelogenin-ELP fusion proteins can be engineered to enhance properties such as elasticity and stability, making them suitable for diverse applications in biomedicine and material science. (Less) - Popular Abstract
- Biomaterials are materials designed to interact with biological systems for medical purposes. They can be made from natural or synthetic sources and are crucial for repairing, replacing, or enhancing damaged tissues and organs. In the realm of biomaterials, proteins derived from natural sources are gaining attention due to their compatibility with biological systems.
Amelogenin is a protein naturally found in dental enamel, the hardest substance in the human body. This protein is remarkable because it can self-assemble into structures called nanospheres and nanoribbons, which are crucial for the proper formation of enamel. Another fascinating protein is the Elastin-Like Polypeptide (ELP), which has the unique ability to change its... (More) - Biomaterials are materials designed to interact with biological systems for medical purposes. They can be made from natural or synthetic sources and are crucial for repairing, replacing, or enhancing damaged tissues and organs. In the realm of biomaterials, proteins derived from natural sources are gaining attention due to their compatibility with biological systems.
Amelogenin is a protein naturally found in dental enamel, the hardest substance in the human body. This protein is remarkable because it can self-assemble into structures called nanospheres and nanoribbons, which are crucial for the proper formation of enamel. Another fascinating protein is the Elastin-Like Polypeptide (ELP), which has the unique ability to change its structure in response to temperature and other environmental conditions. ELPs are highly biocompatible and can be tailored for various applications, such as drug delivery and tissue engineering.
In this study, we aimed to create and analyze new proteins by fusing amelogenin with ELPs. The hypothesis was that these fusion proteins would combine the best properties of both proteins, resulting in new materials with enhanced elasticity and flexibility, ideal for use in medical applications.
Using advanced genetic engineering techniques, we produced eleven different fusion proteins in bacteria. We successfully expressed and purified these proteins, confirming their presence with techniques like SDS-PAGE, which helps to separate proteins based on their size.
We then investigated how these new proteins behave. Dynamic Light Scattering (DLS) revealed that some of these fusion proteins formed nanospheres, much like natural amelogenin, while others formed larger aggregates. Experiments also showed that in the presence of calcium and potassium ions, and under acidic conditions, some of these proteins could form nanoribbons, mimicking the natural assembly process of amelogenin.
Further experiments demonstrated that one of the fusion proteins was particularly good at forming hydrogels, a type of material that can absorb large amounts of water and is useful for medical applications like wound healing. Film formation tests showed that adding certain additives improved the flexibility and mechanical strength of the films made from these proteins.
Overall, this research paves the way for developing new biomaterials with tailored properties, potentially benefiting fields like biomedicine and material science. (Less)
Please use this url to cite or link to this publication:
http://lup.lub.lu.se/student-papers/record/9162527
- author
- Madhu, Anandakrishnan LU
- supervisor
- organization
- course
- KBKM01 20241
- year
- 2024
- type
- H2 - Master's Degree (Two Years)
- subject
- keywords
- Biotechnology, Protein Engineering, Amelogenin- ELP fusion protein, Amelogenin, ELPs
- language
- English
- id
- 9162527
- date added to LUP
- 2024-06-14 13:52:16
- date last changed
- 2024-06-14 13:52:16
@misc{9162527,
abstract = {{This study aimed to engineer and characterize a series of Amelogenin-ELP fusion proteins to explore their potential as biomaterials with enhanced properties. Eleven constructs (176-186) were designed and expressed in E. coli BL21(DE3) cells, with successful purification confirmed by SDS-PAGE. Dynamic Light Scattering (DLS) revealed that constructs 181, 185, and 186 formed nanospheres similar to full-length amelogenin, while constructs 176 showed larger aggregates. Self-assembly experiments demonstrated that construct 183 could form nanoribbons under standard and acidic conditions, with calcium ions playing a crucial role. Hydrogel formation experiments indicated that construct 176 was more prone to gelation, especially in the presence of CaCl₂ and KH₂PO₄, whereas construct 183 showed no gelation, highlighting the significance of engineered modifications. Film formation studies revealed distinct differences based on pH and additives, with additive-enhanced films exhibiting greater flexibility.
The ability of fusion proteins to form hydrogels and films was explored under varying pH and ionic conditions, revealing that specific constructs and conditions could be optimized to create biomaterials with desirable mechanical properties. The formation of nanoribbons and the role of additives in enhancing protein interactions were particularly noted for their potential in creating durable, flexible biomaterials. These findings suggest that Amelogenin-ELP fusion proteins can be engineered to enhance properties such as elasticity and stability, making them suitable for diverse applications in biomedicine and material science.}},
author = {{Madhu, Anandakrishnan}},
language = {{eng}},
note = {{Student Paper}},
title = {{Production and Characterization of Amelogenin-ELP Fusion Proteins}},
year = {{2024}},
}