crystals
Article
Improvement of Organic Soil Shear Strength through Calcite
Precipitation Method Using Soybeans as Bio-Catalyst
Heriansyah Putra 1, * , Erizal 1 , Sutoyo 1 , Minson Simatupang 2






Citation: Putra, H.; Erizal; Sutoyo;
Simatupang, M.; Yanto, D.H.Y.
Improvement of Organic Soil Shear
Strength through Calcite Precipitation
Method Using Soybeans as
Bio-Catalyst. Crystals 2021, 11, 1044.
https://doi.org/10.3390/cryst11091044
Academic Editors: Hamed
Khodadadi Tirkolaei,
Satoru Kawasaki, Liang Cheng and
Leon van Paassen
Received: 27 July 2021
Accepted: 27 August 2021
Published: 30 August 2021
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with regard to jurisdictional claims in
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iations.
Copyright: © 2021 by the authors.
Licensee MDPI, Basel, Switzerland.
This article is an open access article
distributed under the terms and
conditions of the Creative Commons
Attribution (CC BY) license (https://
creativecommons.org/licenses/by/
4.0/).
Crystals 2021, 11, 1044. https://doi.org/10.3390/cryst11091044
and Dede Heri Yuli Yanto 3
1 Department of Civil and Environmental Engineering, IPB University, Bogor 16680, Indonesia;
erizal@apps.ipb.ac.id (E.); sutoyo@apps.ipb.ac.id (S.)
2 Department of Civil Engineering, Halu Oleo University, Kendari 93232, Indonesia;
minson.simatupang@uho.ac.id
3 Research Center for Biomaterials, Indonesian Institute of Sciences (LIPI), Cibinong 16911, Indonesia;
dede@biomaterial.lipi.go.id
* Correspondence: heriansyahptr@apps.ipb.ac.id
Abstract: Organic soil has a high content of water and compressibility. Besides that, it has a low
specific gravity, density, and shear strength. This study evaluates the applicability of the soybean
crude urease for calcite precipitation (SCU-CP) method and its effectiveness in organic soil as a
soil-amelioration technique. Various soybean concentrations were mixed with a reagent composed of
urea and calcium chloride to produce the treatment solution. Its effect on the hydrolysis rate, pH,
and amount of precipitated calcite was evaluated through test-tube experiments. SEM-EDS tests
were performed to observe the mineralogy and morphology of the untreated and treated samples.
The treatment solution composed of the reagent and various concentrations of soybeans was applied
to organic soil. The increasing strength of the organic soil was evaluated using direct shear (DS) and
unconfined compression (UCS) tests. The test-tube results show that a hydrolysis rate of 1600 u/g
was obtained when using 50 g/L of soybeans with a precipitation ratio of 100%. The mechanical
tests show a significant enhancement in the parameters of the organic soil’s shear strength. A shear
strength improvement of 50% was achieved in this study. A UCS of 148 kPa and cohesion of 50 kPa
was obtained in the treated samples of organic soil. This research elucidates that the SCU-CP is an
effective technique for improving organic soil’s shear strength.
Keywords: calcite precipitation; hydrolysis rate; shear strength; soybean; SCU-CP; organic soil
1. Introduction
Organic soil has high compressibility, high saturation, low density, and low shear
strength, resulting in massive and long-term deformation [1,2]. These characteristics bring
about certain problems and limitations for the development of civil structures, especially
in the geotechnical field. Several potential methods have been proposed for improving
the parameters of organic soil, e.g., compaction, deep mixing, stone columns, and the
grouting method using fly ash, cement, sodium silica, and calcite [2–12]. However, the
aforementioned methods involve significant efforts, high costs, and a long span of time
for application, resulting in low efficacy [13]. The utilization of chemicals has also caused
significant environmental impacts, e.g., increasing the soil pH and contaminating the
soil [14–16]. In addition, high viscosity and rapid chemical reactions in the chemical
grouting have been known to reduce the effectivity of this method because the resulting
cement fills the pores and hence, limits the area of improvement [17]. A grouting method
using calcite, called enzyme-induced calcite precipitation (EICP), was introduced as a
promising choice for the soil-enhancement approach [18–22]. The EICP method has been
stated to be able to significantly ameliorate both the shear and the unconfined compressive
strength of the soil. They range from 50 kPa to 1.6 MPa [19,20,23–25]. Moreover, the
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Crystals 2021, 11, 1044 2 of 14

utilization of EICP in sandy soil was reported to have reduced the permeability of the
soil [18,26].
Recently, the EICP method, using a urease enzyme (E1.12) purified from jack
beans, was used as a bio-catalyst in the calcite precipitation method to produce calcite
crystals [18,19]. The utilization of a commercial urease enzyme with this method leads to a
significant financial burden [27,28]. Almajed et al. [28] noted that the utilization of a urease
enzyme purified from the jack bean meal involves high costs. Hence, alternative materials
to replace the commercial urease enzyme should be seriously considered. Some studies
have been performed to look at other sources that can produce urease enzymes, such as
cabbage [27], watermelon [22,29], jack beans [30], and soybeans [31–33]. It was reported
that the utilization of urease harvested from crude soybean is a prospective catalyst, using
the calcite precipitation (CP) approach in refining soil [32,33]. Using soybean crude urease
(SCU), hereafter in this paper referred to as SCU-CP, could upgrade the strength of the
fine-grained soil to twice that of the untreated soil [32]. The improvement in the deviator
strength of the SCU-CP-treated soil varied from 30 kPa to 170 kPa [32].
The calcite precipitation method has been extensively investigated to evaluate its
efficacy in improving the strength of sandy soil [18,22,25,34,35]. However, its utiliza-
tion with organic soil has been very limited. Similar research has been reported by
Sidik et al. [36], Canakci et al. [37], and Chen et al. [38], who used microbial-induced
calcite precipitation to improve organic soil’s shear strength. The amount of enhancement
in the organic soil’s strength was lower than that of the sandy soil [37,38]. Other results
showed an improvement of around 20% in the UCS of the bacterial-based calcite-improved
organic soil [36,39]. The mass of the precipitated calcite within the soil was higher for
the sandy soil than for the organic soil. This was because of the organic and chemical
composition in the organic soil [36,38]. However, the study of the calcite precipitation
method using soybean crude urease as the catalyst to improve the strength of organic soil
has not been discussed in previous studies.
This research aims to assess the applicability of the SCU-CP method to enhance organic
soil strength. The reaction rate and pH change during the urea hydrolysis and their effect
on the production of precipitated calcite are evaluated through test-tube experiments. The
improvement in the organic soil’s shear strength is evaluated using direct shear (DS) and
unconfined compression (UCS) tests. The mineralogical and morphological characteristics
of the precipitated mineral and treated soil samples are also observed using SEM-EDS
(Scanning Electron Microscopy with Energy Dispersive Spectroscopy).

2. Materials and Methods

2.1. Materials
This study observed organic soil collected from Siak, Province of Riau, Indonesia. The
organic soil samples were prepared following Canakci et al. [40]. All the samples were
sieved, and the soil that passed through sieve no. 100 (0.15 mm) was used in this study, as
depicted in Figure 1. Soil property tests were performed to observe the characteristics of the
organic soil. The soil was categorized as organic clay (OH) by the Unified Soil Classification
System (USCS) [41]. It had an organic content of 56.58%, a D10 of 0.11 mm, and a liquid
limit of 126.25%. The grain size distribution and physical properties of the organic soil used
in this study are shown in Figure 2 and Table 1, respectively. In addition, the treatment
solution was prepared by mixing the reagent and the catalyst. The reagent was prepared
by combining the urea and CaCl2 with 95% purity taken from Kanto Chemicals, Tokyo,
Japan. Soybean flour with No. 1031P-BIOCert/LSO-006-IDN/05 /17, obtained from Gasol
Pertanian Organik, was used as the biocatalyst in this study.
 Crystals 2021, 11, 1044 3 of 14
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2021, 11,11, x FOR
x FOR PEER
PEER REVIEW
REVIEW 3 3of of1515

Figure
Figure
Figure1.1.1. Organic
Organic soil
soil
Organic used
used
soil used in
inin this
this
this study.
study.
study.

100
100

8080
Percent finer [%]
Percent finer [%]

6060

4040

2020

00
0.01
0.01 0.10
0.10 1.00
1.00 10.00
10.00
Grainsize
Grain size[mm]
[mm]
Figure
Figure
Figure2.2.2. Grain-size
Grain-size
Grain-size distribution
distribution
distribution of
ofof
thethe
the organic
organic
organic soil
soil
soil used
used
usedininin this
this
this study.
study.
study.

Table
Table
Table1.1.1. Physical
Physical
Physical properties
properties
properties ofofof
thethe
the organic
organic
organic soil
soil
soil used
used
used inin
in this
this
this study.
study.
study.
Property
Property Value
Value Unit
Unit
Property Value Unit
Specific
Specific gravity,
gravity, GsGs 1.61
1.61 -- -
Specific gravity, Gs 1.61
Mean
Mean size,
size,D10
Mean size, D10
D10 0.11
0.11
0.11 mmmm
mm
Uniformity coefficient,CuCuCu
Uniformity
Uniformity coefficient,
coefficient, 4.88
4.88
4.88 -- -
Curvature
Curvature
Curvature coefficient,
coefficient,
coefficient,
Cc Cc
Cc 1.05
1.05
1.05 - -
-
Organic content 56.58 %
Organic
Organic content
content 56.58
56.58 % %
Ash content 43.42 %
Ash
Ash
Liquid content
content
limit, LL 43.42
43.42
126.25 %%%
Liquid limit, LLLL
Liquid
Plastic limit,
limit, PL 126.25
126.25
Non-plastic (NP) %%
%
Plastic
Soil type
Plastic limit,
limit, PLPL Non-plastic
Organic clay (OH)
Non-plastic (NP) (NP) %%
-
Soil
Soil type
type Organic
Organic clay
clay (OH)
(OH) - -
Crystals 2021, 11, 1044 4 of 14

2.2. Analysis of Hydrolysis Rate

Hydrolysis rate determinations took place to assess the efficacy of the soybeans in
changing the urea to carbonate and ammonia ions. This study adopted the research steps
constructed by Whiffin et al. [42]. A standardized curve was prepared by specifying the
conductivity outcoming from the whole hydrolysis of any urea’s concentrations, from
50 mmol/L to 500 mmol/L, that were hydrolyzed by 15 g/L of soybean powder. The
change in the conductivity with time was determined by Hanna Edge Multiparameter
230 and various concentrations of soybean powder, namely, 5, 10, 20, 30, 40, and 50 g/L.
The rate of hydrolysis was measured by computing the conductivity gradient alterations
against time and the curve of the standard curve, expressed in Equation (1).
 
u θms
Hydrolysis rate = ·v· N (1)
g θsc

θ ms is the sample gradient, θ sc is the standardized curve gradient, v is the sample volume
(L), and N is the final ammonia concentration (mmol/L).
pH measurements were also conducted in this study to observe the hydrolysis process
of urea. The determination of the pH evolution with time might indirectly define the
rates and the magnitude of the urea hydrolysis [23]. The tests were conducted using a
pH meter of the Hanna Edge Multiparameter 230. Various concentrations of soybean
solutions were prepared and mixed with 1 mol/L of urea to obtain 100 mL volume in
total. The pH changes were recorded until the pH reached a constant value. During the
measurements, the solution was stirred at a constant speed, and a change in temperature
was also observed.

2.3. Precipitation Test

Tests of precipitation were performed to evaluate the efficacy of soybeans as a cat-
alyst for producing precipitated calcite. The tests adapted the procedures promoted by
Neupane et al. [19]. The reagent was composed of urea and calcium chloride, which were
mixed separately with water, comprising 50% of the final volume of the treatment solution.
The soybean powder was mixed with water for 5 min at a constant speed with a magnetic
stirrer to prepare the other 50% of the final volume of the treatment solution. Thus, the
reagent and soybean solution were mixed thoroughly to make a total volume of 30 mL. The
treatment solution was allowed to react for a curing time of seven days in a transparent
tube at a room temperature of 25 ◦ C. The precipitated mass was evaluated at the end of the
time for curing using filter paper. The filtered treatment solution was then dried in a 60 ◦ C
oven for 24 h. This way, the mass of calcite could be calculated.

2.4. Soil Treatment

Soil treatment was conducted to evaluate the improvement in the soil shear strength
parameters of the organic soil after treating it with a mixed solution consisting of the
reagent and soybean powder. The strength of the treated soil was assessed using a strength
test of both unconfined compressive and direct shear, UCS and DS, respectively. The
organic soil samples were prepared and combined with a controlled volume of treatment
solution. The UCS test samples were created by the compaction of each soil sample into
five layers with a total height and diameter of 10 cm and 5 cm, respectively, to control the
uniformity of the density of the sample. The direct shear test samples were prepared in
two layers with a total height of 2 cm and a diameter of 6 cm. Then, the treated samples
were cured for seven days. A dry density of 0.6 g/cm3 and a degree of saturation of 100%
controlled the mass of each soil sample and the volume of the treatment solution mass.
pH measurements were also performed to evaluate the change in pH of the improved
samples. This research followed the experimental procedures from ASTM D2976 [43].
Treated samples were collected from the UCS and direct shear tests. At various concentra-
tions of soybeans, 3 g of treated soil were prepared and mixed with 50 mL of the calcium
chloride solution with a concentration of 0.01 mol/L for 30 min. The evolution of pH was
Crystals 2021, 11, x FOR PEER REVIEW 5 of 15
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Crystals 2021, 11, 1044 5 of 14

calcium chloride solution with a concentration of 0.01 mol/L for 30 min. The evolution of
calcium chloride solution with a concentration of 0.01 mol/L for 30 min. The evolution of
pH was observed using the Hanna Edge Multiparameter 230 until the pH reached a con-
pH was observed using the Hanna Edge Multiparameter 230 until the pH reached a con-
stant value.
observed SEM-EDS
using (Scanning
the Hanna Electron Microscopy
Edge Multiparameter 230 untilwith
theEnergy
pH reached Dispersive Spectros-
a constant value.
stant value. SEM-EDS (Scanning Electron Microscopy with Energy Dispersive Spectros-
copy)
SEM-EDS analyses were
(Scanning also performed
Electron to
Microscopy assess the
with mineral
Energy composition,
Dispersive the shape
Spectroscopy) of the
analy-
copy) analyses were also performed to assess the mineral composition, the shape of the
precipitated
ses were alsomaterial,
performed andto itsassess
characteristics
the mineral in the organic soil
composition, the samples.
shape of the precipitated
precipitated material, and its characteristics in the organic soil samples.
material, and its characteristics in the organic soil samples.
3. Results and Discussion
3. Results and Discussion
3. Results
3.1. andRate
Hydrolysis Discussion
3.1.
3.1. Hydrolysis
Hydrolysis Rate
Rate
A hydrolysis rate analysis was conducted to evaluate the efficacy of soybeans in hy-
A
A hydrolysis
hydrolysis rate
rate analysis
analysis was
was conducted
conducted to
toevaluate
evaluate the
theefficacy
efficacy of soybeans in
inhy-
drolyzing urea. Various concentrations of urea, namely, 50, 100, 200, 300,of soybeans
and 500 mmol/L, hy-
drolyzing
drolyzing urea.
urea. Various
Various concentrations
concentrations of
of urea,
urea, namely,
namely, 50, 100,
50, 100, 200,
200, 300,
300, and
and 500
500 mmol/L,
mmol/L,
were compounded with 15 g/L of soybeans. The evolution of the conductance was meas-
were compounded
wereuntil
compounded with 15
withwas15 g/L
g/L ofofsoybeans.
soybeans. The evolution of the conductance was meas-
ured all of the urea hydrolyzed. This The
was evolution
indicated of bythethe conductance
conductance was mea-
reaching
ured
sured until all of
until all ofthe urea
the urea was
was hydrolyzed. This was indicated by the conductance reaching
the maximum value. Next, thehydrolyzed. This was indicated
maximum conductance valuesby the conductance
were reaching
plotted to produce a
the maximum value. Next, the maximum conductance values were plotted to produce a
standard curve for hydrolysis, as depicted in Figure 3. Then, 1 mol of urea was prepareda
the maximum value. Next, the maximum conductance values were plotted to produce
standard
standard curve for hydrolysis, as
as depicted in
in Figure 3.
3. Then, 1 mol of of urea was prepared
and mixedcurve
with for hydrolysis,
various concentrationsdepicted Figure
of soybean powderThen,of 15,mol
10, 20, urea was
30, 40, andprepared
50 g/L.
and
and mixed
mixed with
with various
various concentrations
concentrations of
of soybean
soybean powder
powder of
of 5,
5, 10,
10, 20,
20, 30,
30, 40, and
40, and 50 50 g/L.
g/L.
The conductance measurements were performed for 10 min, and the results were plotted
The
The conductance
conductance measurements
measurements were were performed
performed for for 10
10 min,
min, and
and thethe results
results were
were plotted
plotted
on a curve for the measured conductance. The change in conductance with time is shown
on
on aa curve
curve for
for the
the measured
measured conductance.
conductance. The The change
change in in conductance
conductance with with time
time isis shown
shown
in Figure 4.
in
in Figure
Figure 4.4.
45
45
2
40 y = 2.5662 + 0.083225x R 2= 0.996
40 y = 2.5662 + 0.083225x R = 0.996
[mS]
Conductance[mS]

35
35
30
30
Conductance

25
25
20
20
15
15
10
10
5
50 100 200 300 400 500 600
0 100 200 300 400 500 600
Urea [mMol/L]
Urea [mMol/L]
Figure 3. Standard curve of hydrolysis.
Figure
Figure 3.
3. Standard
Standard curve
curve of
of hydrolysis.
hydrolysis.

2 2
S: 5 g/L; : 0.096x; R : 0.999 S: 30 g/L; : 0.373x; R : 0.992
2 2
S: 5 g/L; : 0.096x; R :2 0.999 S: 30 g/L; : 0.373x; R2 : 0.992
S: 10 g/L;  0.156x; R : 0.978 S: 40 g/L;: 0.460x; R : 0.996
2 2
S: 10 g/L;  0.156x; R2 : 0.978 S: 40 g/L;: 0.460x; R2 : 0.996
S: 20 g/L; : 0.270x; R : 0.985 S: 50 g/L; : 0.534x; R : 0.979
2 2
8.0 S: 20 g/L; : 0.270x; R : 0.985 S: 50 g/L; : 0.534x; R : 0.979
[mS]

8.0
conductance[mS]

7.0
7.0
6.0
Measuredconductance

6.0
5.0
5.0
4.0
4.0
3.0
3.0
Measured

2.0
2.0
1.0
1.0
0.0
0.00.0 2.0 4.0 6.0 8.0 10.0
0.0 2.0 4.0 6.0 8.0 10.0
Elasped time [minutes]
Elasped time [minutes]
Figure 4. Change in conductance with time for various concentrations of soybeans.
Crystals 2021, 11, x FOR PEER REVIEW 6 of 15

Crystals 2021, 11, 1044 6 of 14

Figure 4. Change in conductance with time for various concentrations of soybeans.

The conductance measurements were performed for various concentrations of soy-

beans,Theresulting in a linear
conductance curve withwere
measurements a slope. The slopes
performed of the curves
for various were thenofcom-
concentrations soy-
pared to the standard curve to obtain the hydrolysis rate depicted
beans, resulting in a linear curve with a slope. The slopes of the curves were then in Figure 5. compared
As is ap-
parent, the activity
to the standard was
curve to linearly related
obtain the to the rate
hydrolysis increase in soybean
depicted in Figureconcentration. The hy-
5. As is apparent, the
drolysis rate linearly
activity was of 1600 related
u/g (noteto that 1 u of activity
the increase corresponds
in soybean to 1 μmol/L
concentration. of urea hydro-
The hydrolysis rate of
lyzed per (note
1600 u/g minute)thatwas
1 u obtained
of activityusing 50 g/L of
corresponds tosoybeans.
1 µmol/L On average,
of urea the useper
hydrolyzed of 1minute)
g/L of
soybeans was estimated to have an activity of around 40 u/g. Compared
was obtained using 50 g/L of soybeans. On average, the use of 1 g/L of soybeans was to commercial
enzymes
estimatedthat have an
to have an activity
activity of
of 2950
around u/g 40
foru/g.
15 g/L (or around
Compared to200 u/g for 1 g/L
commercial of urease)
enzymes that
or
have an activity of 2950 u/g for 15 g/L (or around 200 u/g for 1 g/L of urease) orofother
other plant-derived ureases (i.e., watermelon [22], cabbage, soy pulp [27]), 5 g/L soy-
beans is high enough
plant-derived to bewatermelon
ureases (i.e., used instead of cabbage,
[22], commercial soyurease enzymes
pulp [27]), 5 g/Lwith the EICP
of soybeans is
method.
high enough to be used instead of commercial urease enzymes with the EICP method.

1600

1400
Hidrolysis rate [u/g]

1200

1000

800

600

400

200

0
5 10 20 30 40 50
Soybean [g/L]
Figure
Figure5.
5. Hydrolysis
Hydrolysisrate
rateresults.
results.
Crystals 2021, 11, x FOR PEER REVIEW 7 of 15
The pH
The pH measurements
measurements were
were taken
taken to
to observe
observe the
the elapsed
elapsed time
time of
of the
the hydrolysis
hydrolysis of
of
urea using soybeans. The results of these measurements are depicted in Figure
urea using soybeans. The results of these measurements are depicted in Figure 6. 6.

9.20

9.10
Measured pH [-]

9.00

8.90

8.80

8.70 5 g/L
10 g/L
8.60 20 g/L
30 g/L
8.50 40 g/L
50 g/L
8.40
0 5 10 15 20 25 30
Elapsed time [minutes]
Figure
Figure6.6.pH
pHmeasurements
measurementswith
withtime
timefor
forvarious
variousconcentrations
concentrationsof
ofsoybeans.
soybeans.

The results show a significant improvement in the pH value at the beginning of mix-
ing. It reached a constant value after 30 min, in the range of 9.08 to 9.12, depending on the
soybean concentration. The increasing pH indicates the change in urea to carbonate and
ammonia ions by means of hydrolysis. The released ammonia (NH4) ions promoted the
increase in pH [44]. Furthermore, the constant value of the pH confirms the completion of
Crystals 2021, 11, 1044 7 of 14

The results show a significant improvement in the pH value at the beginning of mixing.
It reached a constant value after 30 min, in the range of 9.08 to 9.12, depending on the
soybean concentration. The increasing pH indicates the change in urea to carbonate and
ammonia ions by means of hydrolysis. The released ammonia (NH4 ) ions promoted the
increase in pH [44]. Furthermore, the constant value of the pH confirms the completion of
the reaction. The entire amount of urea was converted to carbonate and ammonia ions. The
results of the pH measurements also show a decrease in the activity of the enzymes when
the pH was 9 [45]. In addition, the pH results indicate that the concentration of soybeans
had no significant impact on the pH. A relatively similar pH was obtained for the various
concentrations of soybeans, ranging from 5 to 50 g/L, resulting in pH levels ranging from
9.08 to 9.12.

3.2. Precipitation Test

The precipitation tests were performed to evaluate the efficacy of the SCU-CP method
by measuring the mass of the produced calcite. This test adopts the experimental procedure
developed by Neupane et al. [19]. First, soybean and reagent solutions were prepared sepa-
rately. Next, the reagent composed of urea and calcium chloride, each with concentrations
of 0.5, 1.0, and 1.5 mol/L, and soybeans with various concentrations of 5 to 50 g/L, were
prepared and mixed with distilled water separately. Then, 30 mL total volume of both
soybean and reagent solution were mixed in a transparent tube and cured for three days.
The conditions of the precipitation tests are depicted in Table 2. The precipitated materials
were evaluated after the curing times. Firstly, the treatment solution was filtrated using
filter paper No. 41 (pore size of 20 µm) to collect the material deposits. Next, the retained
material was dried in an oven at 60 ◦ C for 24 h to enable the evaluation of the dry mass.
Then, its productivity was examined as percentages of the precipitated material’s dry mass,
and the maximum theoretical mass for each concentration was calculated.

Table 2. Condition of the precipitation test.

Case Concentration of Reagent (mol/L) Concentration of Soybeans (g/L)

S1.1 0.50 5
S1.2 0.50 10
S1.3 0.50 20
S1.4 0.50 30
S1.5 0.50 40
S1.6 0.50 50
S2.1 1.00 5
S2.2 1.00 10
S2.3 1.00 20
S2.4 1.00 30
S2.5 1.00 40
S2.6 1.00 50
S3.1 1.50 5
S3.2 1.50 10
S3.3 1.50 20
S3.4 1.50 30
S3.5 1.50 40
S3.6 1.50 50

The productivity of the various concentrations of soybeans and reagents in producing

a precipitated material is shown in Figure 7. The precipitation ratio significantly increased
when the concentration of soybean was increased from 5 to 20 g/L. Then, the precipitation
mass reached a stable condition even when the soybean concentrations were enhanced. The
increase in soybean concentration was seen to promote an improvement in the hydrolysis
rates, and thus, it enhanced the calcite mass. In addition, the various concentrations
of the reagent resulted in a different trend in the calcite mass. The increasing reagent
Crystals 2021, 11, 1044 8 of 14

concentration brought about a lower precipitation ratio in the same soybean concentration.
These results indicate that soybean concentration is a crucial parameter in this method.
Hence, the optimum concentration of reagent and soybeans should be considered. The
increase in the concentration of the reagent from 0.5 to 1.5 mol/L should be followed by the
enhancement of the concentration of soybean from 5 to 30 g/L. Neupane et al. [19] noted
that the upgrade in urease concentration had a notable influence on the precipitation ratio
in the calcite precipitation method. Furthermore, this study shows that a high concentration
of soybeans was able to inhibit the precipitation process. The precipitation ratio decreased
from 90 to 95% when 40 to 50 g/L of soybeans were appended to the solution. The organic
content may affect this phenomenon, resulting in undissolved soybeans, hampering
Crystals 2021, 11, x FOR PEER REVIEW 9 of the
15
precipitation process. Hence, a method to isolate the undissolved soybeans should be
considered in order to optimize the use of soybeans as a bio-catalyst.

100
Precipitation ratio [%]

80

60

40

R: 0.5 mol/L
20 R: 1.0 mol/L
R: 1.5 mol/L
0
0 10 20 30 40 50
Soybean [g/L]
Figure
Figure7.7.Precipitation
Precipitationtest
testresults
resultswith
withvarious
variousconcentrations
concentrationsof
ofsoybeans
soybeansand
andreagent.
reagent.

3.3. Direct Shear (DS) Test

3.3. Direct Shear (DS) Test
DS tests were established to assess the effectiveness of SCU-CP on the organic soil’s
DS tests were established to assess the effectiveness of SCU-CP on the organic soil’s
cohesion and internal friction angle. A reagent solution composed of 1.0 mol/L of urea and
cohesion and internal friction angle. A reagent solution composed of 1.0 mol/L of urea and
1.0 mol/L of calcium chloride was prepared and then mixed thoroughly with the selected
1.0 mol/L solutions
soybean of calciumofchloride was30
10, 20, and prepared
g/L. Each andsoil
then mixedwas
sample thoroughly
mixed with withthethetreatment
selected
soybean solutions of 10, 20, and 30 g/L. Each soil sample was mixed
solution, and thus, the mold sample was formed. The soil sample was controlled under with the treatment
solution,
saturatedand thus, the
conditions mold
with sample
a dry was
density of formed.
0.6 g/cmThe soiluntreated
3 . The sample was controlled
soil samples under
were also
saturated conditions with a dry density of 0.6 g/cm 3. The untreated soil samples were also
prepared in the same way using a controlled volume of water. All the samples were cured
prepared
for sevenindays,
the same way DS
and then using a controlled
tests volumeunder
were conducted of water. All the
vertical samples
stress in thewere
ordercured
of 50,
for seven days,
100, and 150 kPa. and then DS tests were conducted under vertical stress in the order of 50,
100, and 150 kPa.
The results of the DS tests on the organic soil treated by the calcite precipitation
The results
solution of the DS
with various tests on
soybean the organic soil
concentrations treated byinthe
are depicted calcite
Figure precipitation
8. The results showso-
lution with various soybean concentrations are depicted in Figure 8.
that the cohesion levels of the improved soil were augmented significantly. Improvements The results show that
the cohesion levels
in cohesion, ranging of the
fromimproved
43 to 79% soil were augmented
compared significantly.
to the untreated Improvements
soil, were obtained for in
cohesion,
the various ranging
soybean from 43 to 79% compared
concentrations. Increasing to the
theuntreated soil, were obtained
soybean concentration also ledfortothe
an
various soybean
improvement in concentrations. Increasing
cohesion. The treatment the soybean
solution composedconcentration also and
of the reagent led to an im-
soybeans
provement
was able toinpromote
cohesion. The precipitation
calcite treatment solution
and the composed of the of
strengthening reagent
the soil and soybeans
particles. In
was able to promote calcite precipitation and the strengthening
comparison, the improvement in the strength gained in this research was significantly of the soil particles. In
comparison,
higher than the thatimprovement
in bacterial-basedin thecalcite
strength gained in techniques
precipitation this research was significantly
[36,38]. The results
higher
confirmthan that
that thein application
bacterial-based calcite precipitation
of soybeans might be more techniques [36,38].
effective thanThe results con-
bacteria. This
firm
couldthat
bethedueapplication of soybeans
to the activity might bebacteria
of susceptible more effective than bacteria.
and influenced This could be
by environmental
due to the activity of susceptible bacteria and influenced by environmental conditions
[46]. In addition, it was shown that a lower pH hampered the bacterial activity, thus re-
ducing the hydrolysis process [47].
Furthermore, the treatment of organic soil using precipitation was seen to have no
significant effect on the internal friction angle. The precipitated material within the soil
Crystals 2021, 11, 1044 9 of 14

Crystals 2021, 11, x FOR PEER REVIEW 10 of 15

conditions [46]. In addition, it was shown that a lower pH hampered the bacterial activity,
thus reducing the hydrolysis process [47].

Figure8.8.Direct
Figure Directshear
sheartest
testresults
resultswith
withvarious
variousconcentrations
concentrationsofofsoybeans.
soybeans.

Furthermore,
3.4. Unconfined the treatment
Compressive of organic
Strength (UCS)soil using precipitation was seen to have no sig-
nificant effect on the internal friction angle. The precipitated material within the soil caused
Samples for the UCS tests were set up and treated in a similar way to the DS tests.
a slight decrease in the friction of the soil. Putra et al. [48] reported that the precipitated ma-
terial within the soil of
The augmentation the organicpromotes
significantly soil’s compressive
bonds among strength
the soilatparticles,
various soybean concen-
thus improving
trations
the bondingis depicted
and cohesion in Figure
of the9.soil,
Thehenceresults show that
decreasing theapplying the calcite
friction angle precipitation
of the soil particles.
technique resulted in a significant improvement in the UCS.
Wibisono et al. [49] also reported that clay in sandy soil brings about high cohesion Strength ranging from 130 andto
148 kPa or a gain of 37 to 58% was obtained in this research compared
significantly reduces the internal friction angle. In addition, Asghari et al. [50] reported to the untreated
soil.the
that Thecementation
variation inprocess
soybean hasconcentration
no prominentalso promoted
impact on the ainternal
slight increase in the UCS.
friction angle.
The rise in the strength of soil observed in this research was relatively lower than that of
sandy
3.4. soil. Putra
Unconfined et al. [24]Strength
Compressive reported(UCS) that using a grouting solution composed of 2 g/L of
the urease
Samples for the UCS tests were set reagent
enzyme and 1 mol/L of the resultedinin
up and treated an improvement
a similar way to theinDS the strength
tests. The
of 205 kPa. This lower improvement in strength may cause a
augmentation of the organic soil’s compressive strength at various soybean concentrations lower pH of 3 to 4 and or-
isganic content
depicted in the 9.
in Figure soilThesample.
resultsStocks-Fischer
show that applying et al. [51]
the reported that the optimum
calcite precipitation technique pH
for calcite promotion in calcite precipitation is 8.3 to 9.0.
resulted in a significant improvement in the UCS. Strength ranging from 130 to 148 kPaPrevious studies also reported
orthat a pHofof
a gain 377.0 is thewas
to 58% optimum
obtained condition for precipitated
in this research compared calcite
to the[23]. In addition,
untreated soil. Thethe
presence in
variation of soybean
the organic content coming
concentration from the asoil
also promoted sample
slight and in
increase thethe
undissolved
UCS. The rise soy-
inbeans may hamper
the strength theobserved
of soil precipitation
in thisprocess; thus,
research wastherelatively
production of calcite
lower is limited
than that [27].
of sandy
The results of the DS tests show a similar trend to that of the
soil. Putra et al. [24] reported that using a grouting solution composed of 2 g/L of the UCS tests conducted in this
research. The UCS improvement also demonstrates an increase
urease enzyme and 1 mol/L of the reagent resulted in an improvement in the strength of in cohesion. These results
confirm
205 that the
kPa. This precipitated
lower improvement calcite enhancedmay
in strength the cause
cohesion of the
a lower pHsoil
ofparticles
3 to 4 and [52].
organic
content in the soil sample. Stocks-Fischer et al. [51] reported that the optimum pH for
calcite promotion in calcite precipitation is 8.3 to 9.0. Previous studies also reported that a
pH of 7.0 is the optimum condition for precipitated calcite [23]. In addition, the presence
of the organic content coming from the soil sample and the undissolved soybeans may
hamper the precipitation process; thus, the production of calcite is limited [27]. The results
of the DS tests show a similar trend to that of the UCS tests conducted in this research. The
UCS improvement also demonstrates an increase in cohesion. These results confirm that
the precipitated calcite enhanced the cohesion of the soil particles [52].
Crystals 2021, 11, x FOR PEER REVIEW 11 of 15
Crystals
Crystals 2021,
2021, 11,
11, x1044
FOR PEER REVIEW 1110ofof 15
14

180
180 Untreated soil Treated soil: CaCl2: 1 mol/L; CO(NH2)2: 1 mol/L
160 Untreated soil Treated soil: CaCl2: 1 mol/L; CO(NH2)2: 1 mol/L
160
140
140

[kPa]
120

[kPa]
120
100
UCS 100
80
UCS
80
60
60
40
40
20
20
0
0 0 10 20 30
0 10 20 30
Soybean [g/L]
Soybean [g/L]
Figure 9. Results of UCS tests with various concentrations of soybeans.
Figure
Figure9.
9. Results
Resultsof
ofUCS
UCStests
testswith
withvarious
variousconcentrations
concentrations of
of soybeans.
soybeans.
3.5. pH Measurement
3.5. pH Measurement
3.5. pH
TheMeasurement
pH measurement results are presented in Figure 10. The untreated organic soil
The pH measurement results are presented in Figure 10. The untreated organic soil
The pH
was highly measurement
acidic, with a pHresults
of 3.4. are
Thepresented
soil treatment in Figure 10. in
resulted The untreated
a slight organic soil
improvement in
was highly acidic, with a pH of 3.4. The soil treatment resulted in a slight improvement
waswith
pH highly acidic, with
a maximum pHa of
pH5.9.
of 3.4.
The The
lowersoilimprovement
treatment resulted
in pH in maya slight
have improvement
been due to the in
in pH with a maximum pH of 5.9. The lower improvement in pH may have been due to
pH
fact with a maximum pH of 5.9. The lower improvement in pH may have been due to the
the that the precipitated
fact that solution
the precipitated was was
solution a weak basebase
a weak material withwith
material a maximum
a maximum pH pH
of 9,of
fact that the
whereas thesoil
precipitated
sample wassolution
stronglywas a weak
acidic base
[51]. Thismaterial
condition with
may a also
maximum
have pHa of
had sig-9,
9, whereas the soil sample was strongly acidic [51]. This condition may also have had a
whereaseffect
nificant the soil
on sample
the was strongly
precipitation acidic
process. A [51].
lower This
pH condition
during themay also have
treatment had amay
process sig-
significant effect on the precipitation process. A lower pH during the treatment process
nificant
have effect on
thethe precipitation process. A lower pH during theformation
treatmentand
process may
mayinhibited
have inhibited hydrolysis process
the hydrolysis and thus,
process hampered
and calcite
thus, hampered reduced
calcite formation and
have
the inhibited
calcitethe
reduced the
content hydrolysis
calcite[51]. process
content [51]. and thus, hampered calcite formation and reduced
the calcite content [51].
7.0
7.0 Untreated soil Treated soil: CaCl2: 1 mol/L; CO(NH2)2: 1 mol/L
6.0 Untreated soil Treated soil: CaCl2: 1 mol/L; CO(NH2)2: 1 mol/L
6.0
[-][-]

5.0
pHpH

5.0
4.0
Measured

4.0
Measured

3.0
3.0
2.0
2.0
1.0
1.0
0.0
0.0 0 10 20 30
0 10 20 30
Soybean [g/L]
Soybean [g/L]
Figure
Figure10.
10.pH
pHmeasurements
measurementsofofthe
theorganic
organicsoil.
soil.
Figure 10. pH measurements of the organic soil.
3.6. Scanning Electron Microscopy with Energy Dispersive Spectroscopy (SEM-EDS)
3.6. Scanning Electron Microscopy with Energy Dispersive Spectroscopy (SEM-EDS)
3.6. Scanning
SEM-EDSElectron
SEM-EDS tests wereMicroscopy
tests were performedwithtoto
performed Energy Dispersive
observe
observe Spectroscopy
the particle
the particle (SEM-EDS)
morphology
morphology
and composition
and composition
of the precipitated material, organic soil, and treated soil. The outputs of the SEM-EDS tests
of theSEM-EDS testsmaterial,
precipitated were performed
organic to observe
soil, the particle
and treated morphology
soil. The and
outputs of composition
the SEM-EDS
are depicted in Figure 11. Figure 11a shows the morphology of the precipitated material and
of theare
tests precipitated
depicted material,
in Figure organic
11. Figuresoil, and
11a treated
shows thesoil. The outputs
morphology of of the
the SEM-EDS
precipitated
its components that were observed using SEM-EDS with a magnitude of 1000 times. The
tests are depicted in Figure 11. Figure 11a shows the morphology of the precipitated
Crystals 2021, 11, x FOR PEER REVIEW 12 of 15

Crystals 2021, 11, 1044 11 of 14

material and its components that were observed using SEM-EDS with a magnitude of 1000
times. The precipitated material was dominated by the amorphous, spherical, and rhom-
bohedral form
precipitated of thewas
material calcium carbonate.
dominated Inamorphous,
by the addition, thespherical,
fiber form,
andwhich is indicated
rhombohedral as
form
of the calcium carbonate. In addition, the fiber form, which is indicated
undissolved soybeans, was obtained from the SEM images. The amorphous and spherical as undissolved
soybeans, was obtained
shape indicates from the SEM
that the precipitation images.
process wasThenotamorphous and spherical
optimum; hence, shape
the calcination
indicates that the precipitation process was not optimum; hence, the calcination
process was hampered [24]. Furthermore, the presence of the organic content, resulting process was
in
hampered [24]. Furthermore, the presence of the organic content, resulting in
undissolved soybeans, may have inhibited the calcination process [27]. Therefore, the ex- undissolved
soybeans, may have
traction method forinhibited
soybean the calcination
powder should process [27]. Therefore,
be considered the extraction
essential method
in the subsequent
for soybean
study. powder should be considered essential in the subsequent study.

(a) Precipitated material

(b) Untreated organic soil

(c) Treated organic soil

Figure 11. SEM-EDS test results: (a) precipitated material, (b) untreated organic soil, and (c) treated organic soil.
Figure 11. SEM-EDS test results: (a) precipitated material, (b) untreated organic soil, and (c) treated organic soil.
The EDS results show that the precipitated material and organic soil had carbon,
The and
oxygen, EDSaluminum.
results showInthat the precipitated
addition, materialmaterial
the precipitated and organic
wassoil
alsohad carbon, ox-
composed of
ygen, and aluminum. In addition, the precipitated material was also composed of
calcium. Meanwhile, the organic soil had silica. Figure 11b shows the SEM-EDS results forcalcium.
Meanwhile,
the unimproved the organic
organicsoil.
soil The
hadsheet
silica.
andFigure
form 11b shows
of the the SEM-EDS
fiber were results
both clearly for the
observed in
the soil sample. Furthermore, Figure 11c shows the SEM-EDS results for the treated organic
Crystals 2021, 11, 1044 12 of 14

soil. The SEM tests were performed with a magnitude of 5000 times. The precipitated
calcium carbonate was seen to be covering the sheet of organic soil. It filled the pores,
bound soil particles, and thus increased the organic soil’s strength. The SEM image clearly
shows that the crystals precipitated on the organic soil’s surface. However, the precipitated
materials promoted in the treated were dominated by the amorphous form. It may also have
impacted the presence of the organic content, resulting in soil samples and undissolved
soybeans. The rhombohedral and spherical shape of the precipitated materials may have
had difficulty forming in the high content of organic materials. The precipitated form and
its location may have caused the lower effectivity of the improvement in the organic soil
compared to that in the sandy soil, where the precipitated material ideally formed as calcite
in the rhombohedral form on the contact surface of the soil particles [40]. Canacki et al. [40]
also reported that in the organic soil treatment by the calcite precipitation method, the
precipitated material formed on the surface and filled the pores of the organic soil.

4. Conclusions
The SCU-CP method was evaluated for its effectiveness as a soil-enhancement tech-
nique. Any experiments were performed to assess the applicability of soybean powder
as a new material for a bio-catalyst. The hydrolysis-rated results confirm that the urease
activity was linearly related to the increase in soybean concentration. A hydrolysis rate of
1600 u/g was obtained using 50 g/L of soybeans. Hence, the soybeans had a high-enough
hydrolysis rate to be used instead of commercial urease enzymes with the calcite precip-
itation method. The precipitation test results show that the soybean concentration is a
crucial parameter in this method. The precipitation ratio significantly increased when the
soybean concentration was increased from 5 g/L to 20 g/L and reached the optimum ratio
of 20 to 30 g/L of soybeans. In addition, the high concentration of soybeans was able to
inhibit the precipitation process. The precipitation ratio dropped from 90 to 95% when 40
to 50 g/L of soybeans were supplemented in the solution. The mechanical tests showed
a significant amelioration in the parameters of the organic soil’s shear strength. A shear
strength improvement of 50% was achieved in this study. A UCS and a cohesion of 148 kPa
and 50 kPa, respectively, were obtained for the treated samples of organic soil. This study
confirms that SCU-CP is a practical approach to improve the organic soil’s shear strength.

Author Contributions: H.P. conducted the experiment work, collected the data, and wrote the paper.
E. and S. suggested the experimental methods and how to analyze the data. M.S. partly suggested the
method and partly wrote the paper. D.H.Y.Y. partly conducted the experimental work. All authors
have read and agreed to the published version of the manuscript.
Funding: This research was funded by the Ministry of Education, Culture, Research, and Technology
of Indonesia, grant number 1/E1/KP.PTNBH/2021.
Acknowledgments: The authors acknowledge the managerial team of the Indonesian Institute of
Science, especially the facilities and the scientific and technical assistance of the Integrated Laboratory
of Bioproducts.
Conflicts of Interest: The authors declare no conflict of interest.

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