Recombinant human enamelin produced in Escherichia coli promotes mineralization in vitro
Halablab, Monalissa LU
; Wallman, Lovisa
and Bonde, Johan
LU
(2024)
In BMC Biotechnology
24(1).
- Abstract
Background: Enamelin is an enamel matrix protein that plays an essential role in the formation of enamel, the most mineralized tissue in the human body. Previous studies using animal models and proteins from natural sources point to a key role of enamelin in promoting mineralization events during enamel formation. However, natural sources of enamelin are scarce and with the current study we therefore aimed to establish a simple microbial production method for recombinant human enamelin to support its use as a mineralization agent. Results: In the study the 32 kDa fragment of human enamelin was successfully expressed in Escherichia coli and could be obtained using immobilized metal ion affinity chromatography purification (IMAC),... (More)
Background: Enamelin is an enamel matrix protein that plays an essential role in the formation of enamel, the most mineralized tissue in the human body. Previous studies using animal models and proteins from natural sources point to a key role of enamelin in promoting mineralization events during enamel formation. However, natural sources of enamelin are scarce and with the current study we therefore aimed to establish a simple microbial production method for recombinant human enamelin to support its use as a mineralization agent. Results: In the study the 32 kDa fragment of human enamelin was successfully expressed in Escherichia coli and could be obtained using immobilized metal ion affinity chromatography purification (IMAC), dialysis, and lyophilization. This workflow resulted in a yield of approximately 10 mg enamelin per liter culture. Optimal conditions for IMAC purification were obtained using Ni2+ as the metal ion, and when including 30 mM imidazole during binding and washing steps. Furthermore, in vitro mineralization assays demonstrated that the recombinant enamelin could promote calcium phosphate mineralization at a concentration of 0.5 mg/ml. Conclusions: These findings address the scarcity of enamelin by facilitating its accessibility for further investigations into the mechanism of enamel formation and open new avenues for developing enamel-inspired mineralized biomaterials.
(Less)
- author
- Halablab, Monalissa
LU
; Wallman, Lovisa
and Bonde, Johan
LU
- organization
- publishing date
- 2024-12
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- Biomineralization, Enamel, Enamel matrix protein, Enamelin, Expression, Optimization, Purification
- in
- BMC Biotechnology
- volume
- 24
- issue
- 1
- article number
- 48
- publisher
- BioMed Central (BMC)
- external identifiers
-
- scopus:85198020602
- pmid:38982413
- ISSN
- 1472-6750
- DOI
- 10.1186/s12896-024-00875-0
- language
- English
- LU publication?
- yes
- id
- 74d24766-604b-43fc-be90-8ab539391efb
- date added to LUP
- 2024-08-26 13:55:40
- date last changed
- 2024-08-27 03:00:04
@article{74d24766-604b-43fc-be90-8ab539391efb,
abstract = {{<p>Background: Enamelin is an enamel matrix protein that plays an essential role in the formation of enamel, the most mineralized tissue in the human body. Previous studies using animal models and proteins from natural sources point to a key role of enamelin in promoting mineralization events during enamel formation. However, natural sources of enamelin are scarce and with the current study we therefore aimed to establish a simple microbial production method for recombinant human enamelin to support its use as a mineralization agent. Results: In the study the 32 kDa fragment of human enamelin was successfully expressed in Escherichia coli and could be obtained using immobilized metal ion affinity chromatography purification (IMAC), dialysis, and lyophilization. This workflow resulted in a yield of approximately 10 mg enamelin per liter culture. Optimal conditions for IMAC purification were obtained using Ni<sup>2+</sup> as the metal ion, and when including 30 mM imidazole during binding and washing steps. Furthermore, in vitro mineralization assays demonstrated that the recombinant enamelin could promote calcium phosphate mineralization at a concentration of 0.5 mg/ml. Conclusions: These findings address the scarcity of enamelin by facilitating its accessibility for further investigations into the mechanism of enamel formation and open new avenues for developing enamel-inspired mineralized biomaterials.</p>}},
author = {{Halablab, Monalissa and Wallman, Lovisa and Bonde, Johan}},
issn = {{1472-6750}},
keywords = {{Biomineralization; Enamel; Enamel matrix protein; Enamelin; Expression; Optimization; Purification}},
language = {{eng}},
number = {{1}},
publisher = {{BioMed Central (BMC)}},
series = {{BMC Biotechnology}},
title = {{Recombinant human enamelin produced in Escherichia coli promotes mineralization in vitro}},
url = {{http://dx.doi.org/10.1186/s12896-024-00875-0}},
doi = {{10.1186/s12896-024-00875-0}},
volume = {{24}},
year = {{2024}},
}