Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

The Extracellular and Intracellular Biomineralization Induced by Bacillus licheniformis DB1-9 at Different Mg/Ca Molar Ratios

Version 1 : Received: 12 October 2018 / Approved: 12 October 2018 / Online: 12 October 2018 (11:56:44 CEST)

A peer-reviewed article of this Preprint also exists.

Han, Z.; Gao, X.; Zhao, H.; Tucker, M.E.; Zhao, Y.; Bi, Z.; Pan, J.; Wu, G.; Yan, H. Extracellular and Intracellular Biomineralization Induced by Bacillus licheniformis DB1-9 at Different Mg/Ca Molar Ratios. Minerals 2018, 8, 585. Han, Z.; Gao, X.; Zhao, H.; Tucker, M.E.; Zhao, Y.; Bi, Z.; Pan, J.; Wu, G.; Yan, H. Extracellular and Intracellular Biomineralization Induced by Bacillus licheniformis DB1-9 at Different Mg/Ca Molar Ratios. Minerals 2018, 8, 585.

Abstract

Biomineralization has become a research hotspot and attracted widespread attention in the field of carbonate sedimentology. In this study, Bacillus licheniformis DB1-9 was used to induce the calcium carbonate precipitation at different magnesium calcium molar ratios in the laboratory to further explore the biomineralization mechanism. Phylogenetic tree shows that the bacteria belongs to Bacillus licheniformis species. The ammonia and carbonic anhydrase can be released by this bacteria, resulting in the pH increase, and the carbonic anhydrase can also promote the hydration reaction of carbon dioxide and subsequently produce the bicarbonate and carbonate ions to elevate the supersaturation of calcium carbonate in the liquid culture medium to facilitate the precipitation of carbonate minerals. The calcites have a shape of rhombohedron, dumbell, and elongation, and aragonite often appears in the form of mineral aggregates, besides that there are also the spherical and the fusiform minerals. FTIR result shows there are some organic functional groups, such as C-O-C and C=O, beside of the characteristic peaks of the calcite and the aragonite, indicating that microbial metabolism is closely related to the mineral formation. The superthin slices of the bacteria analyzed by HRTEM, SAED, EDS and STEM show that the surface and EPS can adsorb a large number of calcium ions and magnesium ions and EPS may act as the nucleation sites, what’s more, the intracellular nanometer-scale sphere areas show the amorphous structures, and the intracellular calcium ions and magnesium ions suggeste that they can be transported from the outside to inside the cell by diffusion along the concentration grade from high to low. This study may provide some references to further understand the biomineralization mechanism induced by microorganisms in the laboratory and the field, and also helps to explore the reason of the transition of calcite sea to aragonite sea in the geological history.

Keywords

Mg/Ca; biominerliazation; nucleation site; Bacillus licheniformis DB1-9; carbonic anhydrase; ammonia

Subject

Environmental and Earth Sciences, Environmental Science

Comments (14)

Comment 1
Received: 16 October 2018
Commenter:
The commenter has declared there is no conflict of interests.
Comment: The new model of carbonate mineralization is very novel.
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Comment 2
Received: 16 October 2018
The commenter has declared there is no conflict of interests.
Comment: == == Heading == ==This study investigates the characteristics of microbially mediated minerals and the specificity of this geobiological process at the cellular and media scale. It shows (i) that a newly isolated microbial strain can lead to mineral precipitation both intracellularly and extracellularly, by changing the solution pH conditions and molecular composition, especially through the production ofenzyme and (ii) how those microbially mediated crystals presents specific characteristics, that can be chemical, morphological.
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Comment 3
Received: 16 October 2018
Commenter: Bin Sun
The commenter has declared there is no conflict of interests.
Comment: this article introduce the Extracellular and Intracellular Biomineralization induced by one strain Bacillus licheniformis . it is attractive and many new analyzic methods are used to investigate the relationship between the biomineraliation and microorganism, and thissubject is still needs to be further studied.
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Comment 4
Received: 16 October 2018
Commenter: Ruirui Meng
The commenter has declared there is no conflict of interests.
Comment: The paper studied the effects of Bacillus licheniformis DB1-9 to its ambient environment, and told the mineralogy and morphology of minerals precipitated induced by Bacillus licheniformis DB1-9 in detial. Furthermore, the mechnism of extracellular and intracellular biomineralization also was described well.
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Response 1 to Comment 4
Received: 18 October 2018
Commenter: Wenwen Yu
The commenter has declared there is no conflict of interests.
Comment: The article have revealed that the biomineralization includes the extracellular biomineralization and intracellular biomineralization, and also deeply investigated the intracellular biomineralization. There are only fewer articles about the intracellular biomineralization. Thus, I think it is an excellent work and the article maybe should be accepted by this journal. If accepted, I think the article maybe could be cited by many researchers.
Comment 5
Received: 18 October 2018
Commenter: Juntong Pan
The commenter has declared there is no conflict of interests.
Comment: The article used their isolated bacteria to induce the calcite and aragonite formation. The biomineralization mechanism has been deeply explored. The result shows that pH increase is caused by the ammonia and carbonic anhydrase. I think this idea is more attractive because it has corrected the original result that pH increase is caused by the ammonia. This article is excellent. They do a good job!
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Comment 6
Received: 18 October 2018
Commenter: Juntong Pan
The commenter has declared there is no conflict of interests.
Comment: I think the work done by them is good. The article is very intresting. The inovative result that pH increase is caused by the combination effect of ammonia and carbonic anhydrate is very attractive. I think if the article could be accepted by this journal there will be a large number of citetations. An excellent article!
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Comment 7
Received: 19 October 2018
Commenter: Jiajia Wang
The commenter has declared there is no conflict of interests.
Comment: This paper describes the formation mechanism of intracellular and extracellular minerals, which is very helpful for my study. In addition, this study may provide some references to further understand the biomineralization mechanism induced by microorganisms in the laboratory and the field, and also helps to explore the reason of the transition of calcite sea to aragonite sea in the geological history.
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Comment 8
Received: 19 October 2018
Commenter: Zhenpeng Bi
The commenter has declared there is no conflict of interests.
Comment: The article is very new and innovative, and it is very important that the mechanism and influence of mineralization were discussed in depth。
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Comment 9
Received: 19 October 2018
Commenter: Guanghe Shan
The commenter has declared there is no conflict of interests.
Comment: The research work is about the biomineralization of carbonates induced by bacteria. The biomineralization mechanism has been deeply studied and some innovative results have also been obtained. I agree with the opinion that Ca and Mg ions can be transported from the outside to inside the cell by diffusion. In fact, the transportation of Ca and Mg ions by different kinds of channels belongs to diffusion. I think their work is very intresting and also is very excellent.
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Comment 10
Received: 19 October 2018
Commenter: Binwei Liu
The commenter has declared there is no conflict of interests.
Comment: This paper is new and practical, and the mechanism and influence of mineralization are discussed furthermore.
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Comment 11
Received: 19 October 2018
Commenter: Na Guo
The commenter has declared there is no conflict of interests.
Comment: I think the article is very intresting.It revealed intracellular mineralization and extracellular mineralization.And the biomineralization mechanism was deeply explored.The experiment used more research methods.So,i think it is excellent.
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Comment 12
Received: 19 October 2018
Commenter: Dan Li
The commenter has declared there is no conflict of interests.
Comment: The data in this article are very abundant and new insights have been put forward on the problem of increasing pH in the surrounding environment that causes mineral formation. The mechanism of microbial induced carbonate formation has been further studied. In addition, this paper also discusses the migration of intracellular and extracellular calcium and magnesium ions. I am very interested for this article and look forward to its publication.
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Comment 13
Received: 19 October 2018
Commenter: tiantian wang
The commenter has declared there is no conflict of interests.
Comment: This paper further explores the biomineralization mechanism and provides a reference for further understanding of the microbial-induced biomineralization mechanisms in the laboratory and on-site. The idea is very novel and good, it is very worth learning the article.
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