Engineered Ureolytic Microorganisms Can Tailor the Morphology and Nanomechanical Properties of Microbial-Precipitated Calcium Carbonate
Abstract
We demonstrate for the first time that the morphology and nanomechanical properties of calcium carbonate (CaCO3) can be tailored by modulating the precipitation kinetics of ureolytic microorganisms through genetic engineering. Many engineering applications employ microorganisms to produce CaCO3. However, control over bacterial calcite morphology and material properties has not been demonstrated. We hypothesized that microorganisms genetically engineered for low urease activity would achieve larger calcite crystals with higher moduli. We compared precipitation kinetics, morphology, and nanomechanical properties for biogenic CaCO3 produced by two Escherichia coli (E. coli) strains that were engineered to display either high or low urease activity and the native producer Sporosarcina pasteurii. While all three microorganisms produced calcite, lower urease activity was associated with both slower initial calcium depletion rate and increased average calcite crystal size. Both calcite crystal size and nanoindentation moduli were also significantly higher for the low-urease activity E. coli compared with the high-urease activity E. coli. The relative resistance to inelastic deformation, measured via the ratio of nanoindentation hardness to modulus, was similar across microorganisms. These findings may enable design of novel advanced engineering materials where modulus is tailored to the application while resistance to irreversible deformation is not compromised.
- Authors:
-
[1];
[1]
- Univ. of Colorado, Boulder, CO (United States)
- Univ. of Colorado, Boulder, CO (United States); National Renewable Energy Lab. (NREL), Golden, CO (United States)
- Publication Date:
- Research Org.:
- National Renewable Energy Laboratory (NREL), Golden, CO (United States)
- Sponsoring Org.:
- U.S. Department of Defense (DOD), Defense Advanced Research Projects Agency (DARPA)
- OSTI Identifier:
- 1572267
- Report Number(s):
- NREL/JA-2700-75264
Journal ID: ISSN 2045-2322
- Grant/Contract Number:
- AC36-08GO28308
- Resource Type:
- Accepted Manuscript
- Journal Name:
- Scientific Reports
- Additional Journal Information:
- Journal Volume: 9; Journal Issue: 1; Journal ID: ISSN 2045-2322
- Publisher:
- Nature Publishing Group
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; genetic engineering; morphology; nanomechanics; calcium carbonate
Citation Formats
Heveran, Chelsea M., Liang, Liya, Nagarajan, Aparna, Hubler, Mija H., Gill, Ryan, Cameron, Jeffrey C., Cook, Sherri M., and Srubar, II, Wil V. Engineered Ureolytic Microorganisms Can Tailor the Morphology and Nanomechanical Properties of Microbial-Precipitated Calcium Carbonate. United States: N. p., 2019.
Web. doi:10.1038/s41598-019-51133-9.
Heveran, Chelsea M., Liang, Liya, Nagarajan, Aparna, Hubler, Mija H., Gill, Ryan, Cameron, Jeffrey C., Cook, Sherri M., & Srubar, II, Wil V. Engineered Ureolytic Microorganisms Can Tailor the Morphology and Nanomechanical Properties of Microbial-Precipitated Calcium Carbonate. United States. https://doi.org/10.1038/s41598-019-51133-9
Heveran, Chelsea M., Liang, Liya, Nagarajan, Aparna, Hubler, Mija H., Gill, Ryan, Cameron, Jeffrey C., Cook, Sherri M., and Srubar, II, Wil V. Fri .
"Engineered Ureolytic Microorganisms Can Tailor the Morphology and Nanomechanical Properties of Microbial-Precipitated Calcium Carbonate". United States. https://doi.org/10.1038/s41598-019-51133-9. https://www.osti.gov/servlets/purl/1572267.
@article{osti_1572267,
title = {Engineered Ureolytic Microorganisms Can Tailor the Morphology and Nanomechanical Properties of Microbial-Precipitated Calcium Carbonate},
author = {Heveran, Chelsea M. and Liang, Liya and Nagarajan, Aparna and Hubler, Mija H. and Gill, Ryan and Cameron, Jeffrey C. and Cook, Sherri M. and Srubar, II, Wil V.},
abstractNote = {We demonstrate for the first time that the morphology and nanomechanical properties of calcium carbonate (CaCO3) can be tailored by modulating the precipitation kinetics of ureolytic microorganisms through genetic engineering. Many engineering applications employ microorganisms to produce CaCO3. However, control over bacterial calcite morphology and material properties has not been demonstrated. We hypothesized that microorganisms genetically engineered for low urease activity would achieve larger calcite crystals with higher moduli. We compared precipitation kinetics, morphology, and nanomechanical properties for biogenic CaCO3 produced by two Escherichia coli (E. coli) strains that were engineered to display either high or low urease activity and the native producer Sporosarcina pasteurii. While all three microorganisms produced calcite, lower urease activity was associated with both slower initial calcium depletion rate and increased average calcite crystal size. Both calcite crystal size and nanoindentation moduli were also significantly higher for the low-urease activity E. coli compared with the high-urease activity E. coli. The relative resistance to inelastic deformation, measured via the ratio of nanoindentation hardness to modulus, was similar across microorganisms. These findings may enable design of novel advanced engineering materials where modulus is tailored to the application while resistance to irreversible deformation is not compromised.},
doi = {10.1038/s41598-019-51133-9},
journal = {Scientific Reports},
number = 1,
volume = 9,
place = {United States},
year = {Fri Oct 11 00:00:00 EDT 2019},
month = {Fri Oct 11 00:00:00 EDT 2019}
}
Free Publicly Available Full Text
Publisher's Version of Record
Other availability
Search WorldCat to find libraries that may hold this journal
Cited by: 30 works
Citation information provided by
Web of Science
Web of Science
Save to My Library
You must Sign In or Create an Account in order to save documents to your library.
Works referenced in this record:
Multiscale mechanical quantification of self-healing concrete incorporating non-ureolytic bacteria-based healing agent
journal, October 2014
- Xu, Jing; Yao, Wu
- Cement and Concrete Research, Vol. 64
Phenotypic and genotypic characteristics associated with biofilm formation in clinical isolates of atypical enteropathogenic Escherichia coli (aEPEC) strains
journal, January 2014
- Nascimento, Heloisa H.; Silva, Lucas EP; Souza, Renata T.
- BMC Microbiology, Vol. 14, Issue 1
Comparative study of carp otolith hardness: Lapillus and asteriscus
journal, May 2013
- Ren, Dongni; Meyers, Marc André; Zhou, Bo
- Materials Science and Engineering: C, Vol. 33, Issue 4
Microbial Diversity and Mineralogical-Mechanical Properties of Calcitic Cave Speleothems in Natural and in Vitro Biomineralization Conditions
journal, February 2018
- Dhami, Navdeep K.; Mukherjee, Abhijit; Watkin, Elizabeth L. J.
- Frontiers in Microbiology, Vol. 9
The Influence of Bacillus pasteurii on the Nucleation and Growth of Calcium Carbonate
journal, July 2006
- Mitchell, Andrew C.; Ferris, F. Grant
- Geomicrobiology Journal, Vol. 23, Issue 3-4
Bacterially mediated mineralization of vaterite
journal, March 2007
- Rodriguez-Navarro, Carlos; Jimenez-Lopez, Concepcion; Rodriguez-Navarro, Alejandro
- Geochimica et Cosmochimica Acta, Vol. 71, Issue 5
Homogeneous nucleation: theory and experiment
journal, February 1998
- Oxtoby, D. W.
- Journal of Physics: Condensed Matter, Vol. 10, Issue 4
Micromechanical properties of single crystal hydroxyapatite by nanoindentation
journal, July 2009
- Saber-Samandari, Saeed; Gross, Kārlis A.
- Acta Biomaterialia, Vol. 5, Issue 6
Bacterially Induced Mineralization of Calcium Carbonate in Terrestrial Environments: The Role of Exopolysaccharides and Amino Acids
journal, May 2003
- Braissant, O.; Cailleau, G.; Dupraz, C.
- Journal of Sedimentary Research, Vol. 73, Issue 3
An improved technique for determining hardness and elastic modulus using load and displacement sensing indentation experiments
journal, June 1992
- Oliver, W. C.; Pharr, G. M.
- Journal of Materials Research, Vol. 7, Issue 06, p. 1564-1583
Bio-mediated soil improvement
journal, February 2010
- DeJong, Jason T.; Mortensen, Brina M.; Martinez, Brian C.
- Ecological Engineering, Vol. 36, Issue 2
Homogeneous nucleation: theory and experiment
journal, September 1992
- Oxtoby, D. W.
- Journal of Physics: Condensed Matter, Vol. 4, Issue 38
Micrographical, minerological and nano-mechanical characterisation of microbial carbonates from urease and carbonic anhydrase producing bacteria
journal, September 2016
- Dhami, Navdeep Kaur; Mukherjee, Abhijit; Reddy, M. Sudhakara
- Ecological Engineering, Vol. 94
Rational Control of Calcium Carbonate Precipitation by Engineered Escherichia coli
journal, November 2018
- Liang, Liya; Heveran, Chelsea; Liu, Rongming
- ACS Synthetic Biology, Vol. 7, Issue 11
Micro-Raman spectroscopy applied to depth profiles of carbonates formed in lime mortar
journal, December 2003
- Martinez-Ramirez, S.; Sanchez-Cortes, S.; Garcia-Ramos, J. V.
- Cement and Concrete Research, Vol. 33, Issue 12
Production of Calcite (Calcium Carbonate) Crystals by Soil Bacteria is a General Phenomenon
journal, December 1973
- Boquet, E.; Boronat, A.; Ramos-Cormenzana, A.
- Nature, Vol. 246, Issue 5434
Synchrotron X-Ray Microanalysis and Imaging of Synthetic Biological Calcium Carbonate in Comparison With Archaeological Samples Originating from the Large Cave of Arcy-sur-Cure (28000-24500 BP, Yonne, France)
journal, September 2013
- Chalmin, Emilie; Reiche, Ina
- Microscopy and Microanalysis, Vol. 19, Issue 6
Amorphous, nanocrystalline and crystalline calcium carbonates in biological materials
journal, November 2010
- Wehrmeister, U.; Jacob, D. E.; Soldati, A. L.
- Journal of Raman Spectroscopy, Vol. 42, Issue 5
Calcium Phosphate Biominerals
journal, January 2002
- Elliott, J. C.
- Reviews in Mineralogy and Geochemistry, Vol. 48, Issue 1
On the relationship between indentation hardness and modulus, and the damage resistance of biological materials
journal, July 2017
- Labonte, David; Lenz, Anne-Kristin; Oyen, Michelle L.
- Acta Biomaterialia, Vol. 57
A mineralogical perspective on the apatite in bone
journal, April 2005
- Wopenka, Brigitte; Pasteris, Jill D.
- Materials Science and Engineering: C, Vol. 25, Issue 2
Fracture Sealing with Microbially-Induced Calcium Carbonate Precipitation: A Field Study
journal, March 2016
- Phillips, Adrienne J.; Cunningham, Alfred B.; Gerlach, Robin
- Environmental Science & Technology, Vol. 50, Issue 7
Influence of fine aggregate characteristics on the rheological properties of mortars
journal, April 2008
- Westerholm, Mikael; Lagerblad, Björn; Silfwerbrand, Johan
- Cement and Concrete Composites, Vol. 30, Issue 4
Mechanical properties of modern calcite- (Mergerlia truncata) and phosphate-shelled brachiopods (Discradisca stella and Lingula anatina) determined by nanoindentation
journal, December 2009
- Merkel, Casjen; Deuschle, Julia; Griesshaber, Erika
- Journal of Structural Biology, Vol. 168, Issue 3
Mechanical behaviour of brushite and hydroxyapatite coatings electrodeposited on newly developed FeMnSiPd alloys
journal, December 2017
- Fornell, J.; Feng, Y. P.; Pellicer, E.
- Journal of Alloys and Compounds, Vol. 729
Softening the ultra-stiff: Controlled variation of Young’s modulus in single-crystal diamond by ion implantation
journal, September 2016
- Battiato, A.; Lorusso, M.; Bernardi, E.
- Acta Materialia, Vol. 116
Crystal Growth of Calcium Carbonate in Hydrogels as a Model of Biomineralization
journal, May 2012
- Asenath-Smith, Emily; Li, Hanying; Keene, Ellen C.
- Advanced Functional Materials, Vol. 22, Issue 14
The effect of porosity and mineral content on the Young's modulus of elasticity of compact bone
journal, January 1988
- Currey, J. D.
- Journal of Biomechanics, Vol. 21, Issue 2
Engineered applications of ureolytic biomineralization: a review
journal, July 2013
- Phillips, Adrienne J.; Gerlach, Robin; Lauchnor, Ellen
- Biofouling, Vol. 29, Issue 6
Controls on the rate of ureolysis and the morphology of carbonate precipitated by S. Pasteurii biofilms and limits due to bacterial encapsulation
journal, April 2012
- Cuthbert, Mark O.; Riley, Michael S.; Handley-Sidhu, Stephanie
- Ecological Engineering, Vol. 41
Microbiological precipitation of CaCO3
journal, October 1999
- Stocks-Fischer, Shannon; Galinat, Johnna K.; Bang, Sookie S.
- Soil Biology and Biochemistry, Vol. 31, Issue 11
Enzyme-accelerated and structure-guided crystallization of calcium carbonate: Role of the carbonic anhydrase in the homologous system
journal, January 2014
- Müller, Werner E. G.; Schlossmacher, Ute; Schröder, Heinz C.
- Acta Biomaterialia, Vol. 10, Issue 1
Nanomechanical properties of nacre
journal, May 2006
- Katti, Kalpana S.; Mohanty, Bedabibhas; Katti, Dinesh R.
- Journal of Materials Research, Vol. 21, Issue 5
Strain improvement of Sporosarcina pasteurii for enhanced urease and calcite production
journal, May 2009
- Achal, V.; Mukherjee, A.; Basu, P. C.
- Journal of Industrial Microbiology & Biotechnology, Vol. 36, Issue 7
Experimental and numerical modeling of bacterially induced pH increase and calcite precipitation in saline aquifers
journal, July 2009
- Dupraz, Sébastien; Parmentier, Marc; Ménez, Bénédicte
- Chemical Geology, Vol. 265, Issue 1-2
The coprecipitation of Sr into calcite precipitates induced by bacterial ureolysis in artificial groundwater: Temperature and kinetic dependence
journal, September 2005
- Mitchell, Andrew C.; Ferris, F. Grant
- Geochimica et Cosmochimica Acta, Vol. 69, Issue 17
Raman spectroscopy of synthetic, geological and biological vaterite: a Raman spectroscopic study: Raman spectroscopy of synthetic, geological and biological vaterite
journal, September 2009
- Wehrmeister, U.; Soldati, A. L.; Jacob, D. E.
- Journal of Raman Spectroscopy, Vol. 41, Issue 2
Strain improvement of Sporosarcina pasteurii for enhanced urease and calcite production
journal, May 2009
- Achal, V.; Mukherjee, A.; Basu, P. C.
- Journal of Industrial Microbiology & Biotechnology, Vol. 36, Issue 7
Micromechanical properties of single crystal hydroxyapatite by nanoindentation
journal, July 2009
- Saber-Samandari, Saeed; Gross, Kārlis A.
- Acta Biomaterialia, Vol. 5, Issue 6
Enzyme-accelerated and structure-guided crystallization of calcium carbonate: Role of the carbonic anhydrase in the homologous system
journal, January 2014
- Müller, Werner E. G.; Schlossmacher, Ute; Schröder, Heinz C.
- Acta Biomaterialia, Vol. 10, Issue 1
On the relationship between indentation hardness and modulus, and the damage resistance of biological materials
journal, July 2017
- Labonte, David; Lenz, Anne-Kristin; Oyen, Michelle L.
- Acta Biomaterialia, Vol. 57
Multiscale mechanical quantification of self-healing concrete incorporating non-ureolytic bacteria-based healing agent
journal, October 2014
- Xu, Jing; Yao, Wu
- Cement and Concrete Research, Vol. 64
Experimental and numerical modeling of bacterially induced pH increase and calcite precipitation in saline aquifers
journal, July 2009
- Dupraz, Sébastien; Parmentier, Marc; Ménez, Bénédicte
- Chemical Geology, Vol. 265, Issue 1-2
Bio-mediated soil improvement
journal, February 2010
- DeJong, Jason T.; Mortensen, Brina M.; Martinez, Brian C.
- Ecological Engineering, Vol. 36, Issue 2
Controls on the rate of ureolysis and the morphology of carbonate precipitated by S. Pasteurii biofilms and limits due to bacterial encapsulation
journal, April 2012
- Cuthbert, Mark O.; Riley, Michael S.; Handley-Sidhu, Stephanie
- Ecological Engineering, Vol. 41
The coprecipitation of Sr into calcite precipitates induced by bacterial ureolysis in artificial groundwater: Temperature and kinetic dependence
journal, September 2005
- Mitchell, Andrew C.; Ferris, F. Grant
- Geochimica et Cosmochimica Acta, Vol. 69, Issue 17
Bacterially mediated mineralization of vaterite
journal, March 2007
- Rodriguez-Navarro, Carlos; Jimenez-Lopez, Concepcion; Rodriguez-Navarro, Alejandro
- Geochimica et Cosmochimica Acta, Vol. 71, Issue 5
Mechanical properties of modern calcite- (Mergerlia truncata) and phosphate-shelled brachiopods (Discradisca stella and Lingula anatina) determined by nanoindentation
journal, December 2009
- Merkel, Casjen; Deuschle, Julia; Griesshaber, Erika
- Journal of Structural Biology, Vol. 168, Issue 3
A mineralogical perspective on the apatite in bone
journal, April 2005
- Wopenka, Brigitte; Pasteris, Jill D.
- Materials Science and Engineering: C, Vol. 25, Issue 2
Comparative study of carp otolith hardness: Lapillus and asteriscus
journal, May 2013
- Ren, Dongni; Meyers, Marc André; Zhou, Bo
- Materials Science and Engineering: C, Vol. 33, Issue 4
Calcite precipitation induced by polyurethane-immobilized Bacillus pasteurii
journal, March 2001
- Bang, Sookie S.; Galinat, Johnna K.; Ramakrishnan, V.
- Enzyme and Microbial Technology, Vol. 28, Issue 4-5
Urease activity in microbiologically-induced calcite precipitation
journal, February 2002
- Bachmeier, Keri L.; Williams, Amy E.; Warmington, John R.
- Journal of Biotechnology, Vol. 93, Issue 2
On the Precipitation of Calcium Carbonate in the Sea by Marine Bacteria, and on the Action of Denitrifying Bacteria in Tropical and Temperate Seas
journal, March 1913
- Drew, G. Harold
- Journal of the Marine Biological Association of the United Kingdom, Vol. 9, Issue 04
Rational Control of Calcium Carbonate Precipitation by Engineered Escherichia coli
journal, November 2018
- Liang, Liya; Heveran, Chelsea; Liu, Rongming
- ACS Synthetic Biology, Vol. 7, Issue 11
Production of Calcite (Calcium Carbonate) Crystals by Soil Bacteria is a General Phenomenon
journal, December 1973
- Boquet, E.; Boronat, A.; Ramos-Cormenzana, A.
- Nature, Vol. 246, Issue 5434
Real-time monitoring of calcification process by Sporosarcina pasteurii biofilm
journal, January 2016
- Harris, Dustin; Ummadi, Jyothir Ganesh; Thurber, Andrew R.
- The Analyst, Vol. 141, Issue 10
Engineered applications of ureolytic biomineralization: a review
journal, July 2013
- Phillips, Adrienne J.; Gerlach, Robin; Lauchnor, Ellen
- Biofouling, Vol. 29, Issue 6
Homogeneous nucleation: theory and experiment
journal, September 1992
- Oxtoby, D. W.
- Journal of Physics: Condensed Matter, Vol. 4, Issue 38
Bacterially Induced Mineralization of Calcium Carbonate in Terrestrial Environments: The Role of Exopolysaccharides and Amino Acids
journal, May 2003
- Braissant, O.; Cailleau, G.; Dupraz, C.
- Journal of Sedimentary Research, Vol. 73, Issue 3
Calcium Phosphate Biominerals
journal, January 2002
- Elliott, J. C.
- Reviews in Mineralogy and Geochemistry, Vol. 48, Issue 1