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学者姓名：赖汉江
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Abstract ：
One-phase-low-pH method is a simple, efficient and easy-to-use biogrouting method for biomineralization based on an Enzyme Induced Carbonate Precipitation (EICP) process. This method utilizes the low-pH biotreatment solution (a mixture of urease solution and cementation solution) to provide a lag period for the biomineralization process, allowing the biotreatment solution to be uniformly distributed within the soil and thereby improving the uniform distribution of calcium carbonate. The existing one-phase-low-pH method uses a low pH urease solution to prepare the biotreatment solution. However, long-term exposure to a low pH environment may result in a decrease in activity or even inactivation of urease, which is not conducive to the practical application of this technology. In this study, a modified one-phase-low-pH method using low pH cementation solution is proposed. Three sets of tests, including urease activity durability tests, solution tests, and sand column treatment tests, were conducted in this study to clarify the necessity and feasibility of the modified method. The test results showed that the acidic environment accelerated the decrease of urease activity over time. This phenomenon would be more pronounced at a lower pH, and urease would be immediately inactive at a pH lower than 4.5. Meanwhile, a high chemical concentration would also lead to a decrease in activity or even inactivation of urease. If urease is active and the initial pH of the biotreatment solution is higher than 4.5, the pH of the biotreatment solution will rapidly rise to a weakly alkaline state and enzyme-induced carbonate precipitation can occur. A biotreatment solution that would produce relatively uniform biomineralization can be prepared by using cementation solution with a pH range of 1.25-3.5 and bacterial urease solution in a volume ratio of 1:1. For the sand column with relatively uniform biomineralization, the pH of the cementation solution (or the initial pH of the biotreatment solution) has a negligible effect on the strength enhancement for similar calcium carbonate content.
Keyword ：
Biomineralization Biomineralization Enzyme induced carbonate precipitation (EICP) Enzyme induced carbonate precipitation (EICP) Low pH cementation solution Low pH cementation solution One-phase-low-pH biogrouting method One-phase-low-pH biogrouting method Urease activity Urease activity
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| GB/T 7714 | Chen, Yi-Wei , Cui, Ming-Juan , Lai, Han-Jiang et al. Modified one-phase-low-pH EICP method using low-pH cementation solution for soil biomineralization [J]. | ACTA GEOTECHNICA , 2025 . |
| MLA | Chen, Yi-Wei et al. "Modified one-phase-low-pH EICP method using low-pH cementation solution for soil biomineralization" . | ACTA GEOTECHNICA (2025) . |
| APA | Chen, Yi-Wei , Cui, Ming-Juan , Lai, Han-Jiang , Zheng, Jun-Jie , Ren, Yu-Xiao . Modified one-phase-low-pH EICP method using low-pH cementation solution for soil biomineralization . | ACTA GEOTECHNICA , 2025 . |
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Abstract ：
Biomineralization based on bacterial enzyme induced carbonate precipitation (BEICP) process is a promising alternative to cement-based ground treatment technology. The bacterial urease used in BEICP process is usually ultrasonic extracted from urease-producing bacteria. To efficiently extract urease with relatively higher activity from bacterial cells, the ultrasonic extraction parameters of urease were optimized in this study. Next, a series of bacterial urease extraction tests and sand column treatment tests were conducted to investigate the effects of vibration amplitude, upper temperature limit, and cooling method on the urease extraction process and biomineralization of sand. The results show that the upper temperature limit is an important factor affecting the extraction efficiency and the activity of the extracted urease solution, and the optimum upper temperature limit is 50 degrees C. The results indicate that increasing vibration amplitude could improve the extraction efficiency, but it hardly affects the urease activity (UA) under the optimal temperature. Continuous cooling could effectively simplify the operation and further improve the efficiency of urease extraction. Under the same urease activity of biotreatment solution, there is no marked difference in calcium carbonate content (CCC) and unconfined compressive strength of biomineralized sand columns prepared by urease solution extracted with different vibration amplitudes and upper temperature limits. The results of this study could provide a reference for application of BEICP technology of urease extraction to large-scale soil treatment. (c) 2025 Institute of Rock and Soil Mechanics, Chinese Academy of Sciences. Published by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/ 4.0/).
Keyword ：
Bacterial enzyme induced carbonate Bacterial enzyme induced carbonate Biomineralization Biomineralization precipitation (BEICP) precipitation (BEICP) Soil improvement Soil improvement Ultrasound Ultrasound Urease extraction Urease extraction
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| GB/T 7714 | Lai, Hanjiang , Chen, Yiwei , Cui, Mingjuan et al. Extraction of high activity bacterial urease and its application to biomineralization of soil [J]. | JOURNAL OF ROCK MECHANICS AND GEOTECHNICAL ENGINEERING , 2025 , 17 (3) : 1847-1861 . |
| MLA | Lai, Hanjiang et al. "Extraction of high activity bacterial urease and its application to biomineralization of soil" . | JOURNAL OF ROCK MECHANICS AND GEOTECHNICAL ENGINEERING 17 . 3 (2025) : 1847-1861 . |
| APA | Lai, Hanjiang , Chen, Yiwei , Cui, Mingjuan , Zheng, Junjie , Chen, Zhibo . Extraction of high activity bacterial urease and its application to biomineralization of soil . | JOURNAL OF ROCK MECHANICS AND GEOTECHNICAL ENGINEERING , 2025 , 17 (3) , 1847-1861 . |
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Abstract ：
An innovative microbial remediation protocol is proposed to overcome critical limitations of conventional microbial-induced carbonate precipitation (MICP) for concrete crack repair. The method integrates pH preconditioning of Sporosarcina pasteurii with a bioadditive-assisted crystallization strategy to address microbial inactivation under highly alkaline conditions, inefficient calcium utilization, and structural instability caused by metastable vaterite formation. Acidification to pH 5.5 preserved 78 % of urease activity at pH 12.5 by stabilizing bacterial zeta potential, while a composite bioadditive composed of polyvinyl alcohol, sodium alginate, and colloidal silica nanoparticles reduced the critical nucleation radius by 29 %, enhancing calcite crystal formation. Mechanical testing showed a 26.8 % increase in flexural strength and an 88.7 % calcium utilization rate, with durability evaluations confirming stable crack sealing over 180 thermal-humidity cycles. Field-scale application to a deteriorated underground garage demonstrated 92 % void-filling efficiency and compressive strength recovery from 28.5 MPa to 41.2 MPa. The developed protocol eliminates the need for carrier materials and reduces carbon emissions, establishing a scalable and sustainable framework for infrastructure rehabilitation. These results highlight the potential of synergistic biological and material strategies for advancing next-generation self-healing concrete technologies. © 2025
Keyword ：
Bending strength Bending strength Bending tests Bending tests Biological materials preservation Biological materials preservation Carbon carbon composites Carbon carbon composites Compression testing Compression testing Compressive strength Compressive strength Cracks Cracks Fracture testing Fracture testing Hardness testing Hardness testing High performance concrete High performance concrete Self compacting concrete Self compacting concrete Self-healing materials Self-healing materials Tensile testing Tensile testing
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| GB/T 7714 | Xu, Wangqing , Lai, Hanjiang , Cui, Mingjuan et al. Synergistic pH-bioadditive strategy for self-healing concrete: Achieving high-efficiency calcite crystallization and sustainable infrastructure rehabilitation [J]. | Construction and Building Materials , 2025 , 484 . |
| MLA | Xu, Wangqing et al. "Synergistic pH-bioadditive strategy for self-healing concrete: Achieving high-efficiency calcite crystallization and sustainable infrastructure rehabilitation" . | Construction and Building Materials 484 (2025) . |
| APA | Xu, Wangqing , Lai, Hanjiang , Cui, Mingjuan , Zheng, Junjie . Synergistic pH-bioadditive strategy for self-healing concrete: Achieving high-efficiency calcite crystallization and sustainable infrastructure rehabilitation . | Construction and Building Materials , 2025 , 484 . |
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There are two major challenges for the use of enzyme-induced carbonate precipitation (EICP)-based soil improvement method: cost and treatment effect. Optimizing the parameters of the treatment solution is one way to enhance the treatment effect and/or reduce the treatment cost. In this study, three key factors: the initial pH (i.e. pH0) of the enzyme solution used to prepare the treatment solution (i.e. the mixture of enzyme solution, CaCl2 and urea), the urease activity of the treatment solution and the concentration of cementation solution (i.e. CaCl2 and urea) are investigated. Crude soybean enzyme solution and the one-phase-low-pH injection method are adopted for the treatment of sand. The results show that the pH0 of the enzyme solution affects the urease activity of enzyme and thus the urease activity of the prepared treatment solution. It is discovered in this paper that there is a threshold pH value for the treatment solution. Only when the pH of the treatment solution is higher than the threshold pH value, calcium ions convert completely into calcium carbonate. There is also a threshold urease activity which is affected by the concentration of cementation solution, CCS. The optimal CCS is 1.0 M. When the CCS is higher than 1.75 M, the urease activity of soybean enzyme solution would be completely lost. These findings are important in guiding the application of EICP treatment using the crude soybean enzyme in real soil improvement projects.
Keyword ：
Crude soybean enzyme Crude soybean enzyme Enzyme-induced carbonate precipitation Enzyme-induced carbonate precipitation Influencing factors Influencing factors Soil improvement Soil improvement
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| GB/T 7714 | Cui, Ming-Juan , Chu, Jian , Lai, Han-Jiang . Optimization of one-phase-low-pH enzyme-induced carbonate precipitation method for soil improvement [J]. | ACTA GEOTECHNICA , 2024 , 19 (3) : 1611-1625 . |
| MLA | Cui, Ming-Juan et al. "Optimization of one-phase-low-pH enzyme-induced carbonate precipitation method for soil improvement" . | ACTA GEOTECHNICA 19 . 3 (2024) : 1611-1625 . |
| APA | Cui, Ming-Juan , Chu, Jian , Lai, Han-Jiang . Optimization of one-phase-low-pH enzyme-induced carbonate precipitation method for soil improvement . | ACTA GEOTECHNICA , 2024 , 19 (3) , 1611-1625 . |
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Abstract ：
Microbially or enzyme induced carbonate precipitation has emerged to be a new type of soil improvement method. However, it appears that the biocementation process is affected by many factors and a common understanding on the control factors on the biocement effect has not been reached. This paper attempts to identify the main factors that controlling the MICP or EICP effect through an in-depth discussion on the fundamentals of biocementation process. Similar to other cemented granular materials, biocemented soil is a structural soil composite consisting of soil skeleton and biocement force chain or biocement network. The strength and stiffness of the biocemented soil is controlled by the reinforcement effect of the biocement network on the soil skeleton or the interplay of the soil skeleton and precipitates. The contribution of the strength by soil skeleton is affected by the soil types and soil properties, while the contribution of the precipitates is through the distribution of the biocement network and the properties of the precipitates. © 2024
Keyword ：
Biocementation Biocementation Influencing factor Influencing factor Mechanism Mechanism Strength enhancement Strength enhancement
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| GB/T 7714 | Lai, H. , Ding, X. , Cui, M. et al. Factors affecting the effectiveness of biocementation of soil [J]. | Biogeotechnics , 2024 , 2 (3) . |
| MLA | Lai, H. et al. "Factors affecting the effectiveness of biocementation of soil" . | Biogeotechnics 2 . 3 (2024) . |
| APA | Lai, H. , Ding, X. , Cui, M. , Zheng, J. , Chu, J. , Chen, Z. . Factors affecting the effectiveness of biocementation of soil . | Biogeotechnics , 2024 , 2 (3) . |
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Calcium salt is an important contributing factor for calcium-based biomineralization.To study the effect of calcium salt on soil biomineralization using crude soybean urease,the calcium salts,including the calcium chloride(CaCl2),calcium acetate((CH3COO)2Ca)and calcium nitrate(Ca(NO3)2),were used to prepare the biotreatment solution to carry out the biomineralization tests in this paper.Two series of biomineralization tests in solution and sand column,respectively,were conducted.Scanning electron microscopy(SEM)and X-ray diffraction(XRD)were performed to determine the microscopic charac-teristics of the precipitated calcium carbonate(CaCO3)crystals.The experimental results indicate that the biomineralization effect is the best for the CaCl2 case,followed by(CH3COO)2Ca,and worst for Ca(NO3)2 under the test conditions of this study(i.e.1 mol/L of calcium salt-urea).The mechanism for the effect of the calcium salt on the biomineralization of crude soybean urease mainly involves:(1)inhibition of urease activity,and(2)influence on the crystal size and morphology of CaCO3.Besides Ca2+,the anions in solution can inhibit the activity of crude soybean urease,and NO3 has a stronger inhibitory effect on the urease activity compared with both CH3COO-and Cl-.The co-inhibition of Ca2+and NO3 on the activity of urease is the key reason for the worst biomineralization of the Ca(NO3)2 case in this study.The dif-ference in biomineralization between the CaCl2 and(CH3COO)2Ca cases is strongly correlated with the crystal morphology of the precipitated CaCO3.
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| GB/T 7714 | Yajie Weng , Junjie Zheng , Hanjiang Lai et al. Biomineralization of soil with crude soybean urease using different calcium salts [J]. | 岩石力学与岩土工程学报（英文版） , 2024 , 16 (5) : 1788-1798 . |
| MLA | Yajie Weng et al. "Biomineralization of soil with crude soybean urease using different calcium salts" . | 岩石力学与岩土工程学报（英文版） 16 . 5 (2024) : 1788-1798 . |
| APA | Yajie Weng , Junjie Zheng , Hanjiang Lai , Mingjuan Cui , Xingzhi Ding . Biomineralization of soil with crude soybean urease using different calcium salts . | 岩石力学与岩土工程学报（英文版） , 2024 , 16 (5) , 1788-1798 . |
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Abstract ：
Calcium salt is an important contributing factor for calcium-based biomineralization. To study the effect of calcium salt on soil biomineralization using crude soybean urease, the calcium salts, including the calcium chloride (CaCl2), calcium acetate ((CH3COO)(2)Ca) and calcium nitrate (Ca(NO3)(2)), were used to prepare the biotreatment solution to carry out the biomineralization tests in this paper. Two series of biomineralization tests in solution and sand column, respectively, were conducted. Scanning electron microscopy (SEM) and X-ray diffraction (XRD) were performed to determine the microscopic characteristics of the precipitated calcium carbonate (CaCO3) crystals. The experimental results indicate that the biomineralization effect is the best for the CaCl2 case, followed by (CH3COO)(2)Ca, and worst for Ca(NO3)(2) under the test conditions of this study (i.e. 1 mol/L of calcium salt-urea). The mechanism for the effect of the calcium salt on the biomineralization of crude soybean urease mainly involves: (1) inhibition of urease activity, and (2) influence on the crystal size and morphology of CaCO3. Besides Ca2+, the anions in solution can inhibit the activity of crude soybean urease, and NO3- has a stronger inhibitory effect on the urease activity compared with both CH3COO- and Cl-. The co-inhibition of C-a2+ and NO3- on the activity of urease is the key reason for the worst biomineralization of the Ca(NO3)(2) case in this study. The difference in biomineralization between the CaCl2 and (CH3COO)2(C)a cases is strongly correlated with the crystal morphology of the precipitated CaCO3. (c) 2024 Institute of Rock and Soil Mechanics, Chinese Academy of Sciences. Production and hosting by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/ licenses/by-nc-nd/4.0/).
Keyword ：
Biomineralization Biomineralization Calcium salt Calcium salt Crude soybean urease Crude soybean urease Influence mechanism Influence mechanism
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| GB/T 7714 | Weng, Yajie , Zheng, Junjie , Lai, Hanjiang et al. Biomineralization of soil with crude soybean urease using different calcium salts [J]. | JOURNAL OF ROCK MECHANICS AND GEOTECHNICAL ENGINEERING , 2024 , 16 (5) : 1788-1798 . |
| MLA | Weng, Yajie et al. "Biomineralization of soil with crude soybean urease using different calcium salts" . | JOURNAL OF ROCK MECHANICS AND GEOTECHNICAL ENGINEERING 16 . 5 (2024) : 1788-1798 . |
| APA | Weng, Yajie , Zheng, Junjie , Lai, Hanjiang , Cui, Mingjuan , Ding, Xingzhi . Biomineralization of soil with crude soybean urease using different calcium salts . | JOURNAL OF ROCK MECHANICS AND GEOTECHNICAL ENGINEERING , 2024 , 16 (5) , 1788-1798 . |
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Abstract ：
Bio-cementation technology based on crude soybean urease is a new environmentally friendly foundation treatment technology emerging in the field of geotechnical engineering. The uniformity of bio-cementation is a pressing issue that needs to be addressed to advance the application of this technology in practical engineering, and soil particle size stands as a significant influencing factor. In this study, 13 types of sand with varying particle sizes were selected, along with self-extracted crude soybean urease solution, to conduct urease percolation tests, sand column curing tests, and scanning electron microscope (SEM) examinations. These experiments aimed to analyze the influence of soil particle size on the effectiveness of bio-cementation using crude soybean urease and explore its underlying mechanisms. The findings reveal that soil particle size significantly affects the migration and adsorption of urease in the crude soybean urease solution. Smaller soil particle sizes facilitate the adsorption of urease. However, excessively small particle sizes (e.g., less than 0.425 mm) lead to the concentration of most adsorbed urease in the middle and upper regions of the soil column. Conversely, excessively large particle sizes (e.g., greater than 4.750 mm) hinder urease adsorption in these regions. Both scenarios tend to result in uneven bio-cementation. Besides the amount of urease adsorption, the influence of soil particle size effect on the biocementation efficacy based on soybean urease is also associated with factors such as pore size within the soil and the number of particle contacts per unit volume of soil. Larger soil particles result in larger interstitial pore sizes and fewer particle contacts, thus hindering the formation of effective calcium carbonate crystals. © 2024 Biodiversity Research Center Academia Sinica. All rights reserved.
Keyword ：
Cementing (shafts) Cementing (shafts) Conservation Conservation Pressing (forming) Pressing (forming) Soil testing Soil testing
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| GB/T 7714 | Lai, Han-Jiang , Liu, Run-Ming , Chen, Zhi-Bo et al. Effect of grain size on biocementation of sand using crude soybean urease [J]. | Rock and Soil Mechanics , 2024 , 45 : 25-32 . |
| MLA | Lai, Han-Jiang et al. "Effect of grain size on biocementation of sand using crude soybean urease" . | Rock and Soil Mechanics 45 (2024) : 25-32 . |
| APA | Lai, Han-Jiang , Liu, Run-Ming , Chen, Zhi-Bo , Cui, Ming-Juan . Effect of grain size on biocementation of sand using crude soybean urease . | Rock and Soil Mechanics , 2024 , 45 , 25-32 . |
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In tropical islands, calcareous sand with poor engineering properties usually needs to be treated before it can be used as building materials. Considering the scarcity of freshwater in these areas, this study proposes seawater-based enzyme induced carbonate precipitation (EICP) technology to enhance the properties of calcareous sand. It is to induce calcium carbonate crystals to bond calcareous sand particles together using the seawater-based crude soybean enzyme and cementation solution (i.e., urea and calcium chloride). In this study, the crude soybean urease extraction test was firstly carried out using seawater and it was also investigated what components of seawater had a greater effect on the soybean urease extraction. Afterwards, the solution test was conducted to explore the ability of the extracted urease in inducing calcium carbonate through analyzing the variation of concentration of calcium ions and pH of the solution. Finally, the biocementation effect of EICP treated calcareous sand using the seawater extracted urease solution was evaluated by the unconfined compressive strength (quc) and microscopic analysis. Test results show that the turbidity of the seawater-extracted soybean urease solution can be reduced by 66.7% compared to that of deionised water extracted urease, with only a slight reduction in urease activity. Among all the components of seawater, NaCl, MgCl2, CaCl2, NaHCO3 and KBr can significantly reduce the turbidity of soybean urease solution. The lower turbidity can effectively avoid bioclogging and contribute to the homogeneity of the EICP-treated calcareous sands, and thus improve the biomineralization efficiency and strength enhancement. Seawater-based EICP treatment will be a great promising technology in freshwater-scarce tropical islands, because it can directly use seawater for biomineralization treatment of calcareous sand, and meanwhile effectively avoid local clogging of biocementation.
Keyword ：
Biomineralization Biomineralization Calcareous sand Calcareous sand Enzyme induced carbonate precipitation (EICP) Enzyme induced carbonate precipitation (EICP) Seawater Seawater Soybean urease extraction Soybean urease extraction
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| GB/T 7714 | Cui, Ming-Juan , Zhou, Jia-Ni , Lai, Han-Jiang et al. Seawater-based soybean urease for calcareous sand biomineralization [J]. | ACTA GEOTECHNICA , 2024 , 19 (10) : 6643-6659 . |
| MLA | Cui, Ming-Juan et al. "Seawater-based soybean urease for calcareous sand biomineralization" . | ACTA GEOTECHNICA 19 . 10 (2024) : 6643-6659 . |
| APA | Cui, Ming-Juan , Zhou, Jia-Ni , Lai, Han-Jiang , Zheng, Jun-Jie , Huang, Ming , Zhang, Zhi-Chao . Seawater-based soybean urease for calcareous sand biomineralization . | ACTA GEOTECHNICA , 2024 , 19 (10) , 6643-6659 . |
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Abstract ：
Enzyme induced carbonate precipitation (EICP) based on self -extracted crude soybean urease solution (CSUS) is a promising method for soil improvement. However, the deionized water -extracted CSUS usually contains a large amount of impurities that easily lead to bioclogging during the biogrouting process, resulting in a nonuniform biomineralization effect. In this study, a purification method using inorganic flocculants is proposed to extract CSUS with relatively high purity and urease activity (UA) for EICP method. Seven commonly used inorganic flocculants were adopted in this study, including KAl(SO4)2 center dot 12H2O, AlCl3 center dot 6H2O, Al2(SO4)3 center dot 18H2O, Fe2(SO4)3, poly aluminum chloride, poly ferric sulfate, and poly aluminum ferric chloride. Three sets of tests, including CSUS extraction tests, solution tests, and sand column treatment tests, were conducted to investigate the feasibility and validity of this purification method. The test results show that inorganic flocculants could effectively reduce the turbidity of the extracted CSUS, avoid the bioclogging during biogrouting, and thus improve the biomineralization effect of the CSUS-based EICP method. Compared with deionized water -based CSUS, at least 60% of the impurities in CSUS can be removed at optimal flocculant contents when the soybean powder content is 100 g/L. On the other hand, the flocculants would also cause a reduction in the UA of the extracted CSUS. Considering the UA, turbidity, and biomineralization effect of the extracted CSUS, the optimal inorganic flocculants and their contents are recommended to be 3.0 g/L for KAl(SO4)2 center dot 12H2O, 2.0 g/L for AlCl3 center dot 6H2O, 2.5 g/L for Al2(SO4)3 center dot 18H2O under the tested conditions, respectively.
Keyword ：
Biomineralization Biomineralization Enzyme induced carbonate precipitation (EICP) Enzyme induced carbonate precipitation (EICP) Inorganic flocculant Inorganic flocculant Soybean urease Soybean urease Urease extraction Urease extraction
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| GB/T 7714 | Lai, Han-Jiang , Liu, Hui , Cui, Ming-Juan et al. Inorganic flocculant-based soybean urease extraction and its effect on biomineralization [J]. | JOURNAL OF CLEANER PRODUCTION , 2024 , 460 . |
| MLA | Lai, Han-Jiang et al. "Inorganic flocculant-based soybean urease extraction and its effect on biomineralization" . | JOURNAL OF CLEANER PRODUCTION 460 (2024) . |
| APA | Lai, Han-Jiang , Liu, Hui , Cui, Ming-Juan , Zheng, Jun-Jie , Chen, Zhi-Bo , Zhang, Zhi-Chao . Inorganic flocculant-based soybean urease extraction and its effect on biomineralization . | JOURNAL OF CLEANER PRODUCTION , 2024 , 460 . |
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