Selective Adsorption of Amino Acids in Crystals of Monohydrocalcite Induced by the Facultative Anaerobic Enterobacter ludwigii SYB1

Yanyang Zhao; Zuozhen Han; Huaxiao Yan; Hui Zhao; Maurice E Tucker; Xiao Gao; Na Guo; Ruirui Meng; Daniel Cosmos Owusu

doi:10.3389/fmicb.2021.696557

Selective Adsorption of Amino Acids in Crystals of Monohydrocalcite Induced by the Facultative Anaerobic Enterobacter ludwigii SYB1

Front Microbiol. 2021 Jul 29:12:696557. doi: 10.3389/fmicb.2021.696557. eCollection 2021.

Authors

Yanyang Zhao^{1

2}, Zuozhen Han^{1

2}, Huaxiao Yan³, Hui Zhao³, Maurice E Tucker^{4

5}, Xiao Gao¹, Na Guo¹, Ruirui Meng¹, Daniel Cosmos Owusu³

Affiliations

¹ Shandong Provincial Key Laboratory of Depositional Mineralization and Sedimentary Minerals, College of Earth Science and Engineering, Shandong University of Science and Technology, Qingdao, China.
² Laboratory for Marine Mineral Resources, Qingdao National Laboratory for Marine Science and Technology, Qingdao, China.
³ Department of Bioengineering, College of Chemical and Biological Engineering, Shandong University of Science and Technology, Qingdao, China.
⁴ School of Earth Sciences, University of Bristol, Bristol, United Kingdom.
⁵ Cabot Institute, University of Bristol, Bristol, United Kingdom.

Abstract

The morphology, crystal structure, and elemental composition of biominerals are commonly different from chemically synthesized minerals, but the reasons for these are not fully understood. A facultative anaerobic bacterium, Enterobacter ludwigii SYB1, is used in experiments to document the hydrochemistry, mineral crystallization, and cell surface characteristics of biomineralization. It was found that carbonate anhydrase and ammonia production were major factors influencing the alkalinity and saturation of the closed biosystem. X-ray diffraction (XRD) spectra showed that calcite, monohydrocalcite (MHC), and dypingite formed in samples with bacterial cells. It was also found that the (222) plane of MHC was the preferred orientation compared to standard data. Scanning transmission electron microscopy (STEM) analysis of cell slices provides direct evidence of concentrated calcium and magnesium ions on the surface of extracellular polymeric substances (EPS). In addition, high-resolution transmission electron microscopy (HRTEM) showed that crystallized nanoparticles were formed within the EPS. Thus, the mechanism of the biomineralization induced by E. ludwigii SYB1 can be divided into three stages: (i) the production of carbonate anhydrase and ammonia increases the alkalinity and saturation state of the milieu, (ii) free calcium and magnesium ions are adsorbed and chelated onto EPS, and (iii) nanominerals crystallize and grow within the EPS. Seventeen kinds of amino acids were identified within both biotic MHC and the EPS of SYB1, while the percentages of glutamic and aspartic acid in MHC increased significantly (p < 0.05). Furthermore, the adsorption energy was calculated for various amino acids on seven diffracted crystal faces, with preferential adsorption demonstrated on (111) and (222) faces. At the same time, the lowest adsorption energy was always that of glutamic and aspartic acid for the same crystal plane. These results suggest that aspartic and glutamic acid always mix preferentially in the crystal lattice of MHC and that differential adsorption of amino acids on crystal planes can lead to their preferred orientation. Moreover, the mixing of amino acids in the mineral structure may also have a certain influence on the mineral lattice dislocations, thus enhancing the thermodynamic characteristics.

Keywords: amino acids; biomineralization; extracellular polymeric substances; facultative anaerobic bacteria; monohydrocalcite.