Startseite Screening and optimization of extracellular pectinase produced by Bacillus thuringiensis SH7
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Screening and optimization of extracellular pectinase produced by Bacillus thuringiensis SH7

  • Maria Umar , Aneela Rehman EMAIL logo , Ibrar Khan , Palwasha Hayat , Azam Hayat , Mujaddad Ur Rehman , Tawaf Ali Shah , Turki M. Dawoud , Safaa Hadrach und Mohammed Bourhia
Veröffentlicht/Copyright: 24. Juli 2023
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Open Chemistry
Aus der Zeitschrift Open Chemistry Band 21 Heft 1

Abstract

The objective of the current research was to identify and evaluate the possibility of production of pectinase, also known as pectin degrading enzymes, from indigenous bacterial strains. Qualitative screening of isolated bacterial strains showed that among 29 bacterial strains, 5 have maximum enzymatic activity. The highest pectinase producing strains were quantitatively analyzed for enzyme production. SH7 strain was found as highest pectinase producer (0.77 IU/mL) that was further analyzed to molecular level by amplification of 16s rRNA. It was found 100% similar with other reported strains of Bacillus thuringiensis. Medium optimization was performed to optimize fermentation conditions for maximum enzyme yield. An experimental design containing 12 experimental runs was designed by Plackett–Burman design (PBD). Maximum pectinase activity was obtained at 45°C after 24 h when the growth medium was supplemented with 2.5% nitrogen, 5.0% substrate, MgSO4 as metal ion, 1% inoculum size, and pH was adjusted to 6. Factorial regression analysis of the PBD design was performed and the overall design was also found significant in terms of R square value. In PBD, the most significant factors for production were temperature, pH, metal ion concentration, and nitrogen source. Central composite design (CCD) design consisting of 26 experimental runs was employed to optimize these four significant factors. The overall model summary showed maximum pectinase activity (19.2 IU/mL) at 37°C temperature, 0.08 NaCl, 1.7% nitrogen source, and pH 8.4. In CCD, NaCl, nitrogen source, and pH were also reported as significant factors by the Pareto chart, probability plots, and 3D interactions.

Keywords: Plackett–Burman design; central composite design; pectinases; optimization; fermentation

1 Introduction

Enzymes are biomolecules that speed up reactions in any metabolic pathway. Many benefits can be acquired by using enzymes that cannot be obtained by traditional ways of treating chemical. They have been more valuable in industrial operations due to their capacity to accomplish very specific chemical transformation. With the passage of time, as the energy consumptions increases, natural resources will be depleted at an alarming rate. Therefore, the need for additional energy resources is critical. There are numerous advantages of applying enzymes over traditional methods [1].

Pectin is a polymeric substance containing group of carbohydrates that esterifies with methyl alcohol. It is a crucial part of the plants’ cell wall. This substance is mostly found in the terrestrial plants and middle plate, where it serves as a binding agent between neighboring cells as well as it also has a role in cell wall hydration. Pectic polymers have been suggested to play a variety of activities, such as cell growth, controlling cell–cell adhesion, wall mechanical characteristics, mediating cell porosity, acting as a source of signaling molecules (oligosaccharides), and taking part in cell organogenesis and differentiation [2]. Pectinases belong to class of enzymes which carried out the cleavage of pectin substances found in the cell walls of plants. Pectic enzymes primarily fall into two categories: those that catalyze the de-esterification of pectins (de-esterifying enzymes) and those that depolymerize glycosidic-(1–4) links within GalA residues (depolymerizing enzymes), through hydrolysis (hydrolases).

There are many sources by which enzymes are synthesized such as plants, animals, and microbes. However, it is well recognized that microorganisms are a key origin of enzymes since they enable inexpensive technique with low waste production and less resource consumption that does not raise any environmental difficulties, as is the case with sources from animals and plants [3]. According to reports, fungi, yeast, and bacteria are responsible for producing 50% of all known enzymes, followed by 35% by bacteria and the remaining 15% by either plants or animals [4]. Many microbial species have been reported for production of extracellular and intracellular pectinases, these include Bacillus subtilis, Bacillus pumilis, Aspergillus niger, Geotrichumklebahnii, Cystofilobasidium capitatum, and Geotrichum lactis [5].

Pectinases can be used in various industrial sectors. The microbial pectinases are being used in the fruit juice industries for the enhancement of clarification and manufacturing of juices. They also minimize the density of fluid and enhance the recovery process [6]. Pectinolytic enzymes also have significant role in wine industry. Wine which is processed enzymatically has better stability and less filtration time as compared to other methods [7]. Many fiber crops are retted and degummed using pectinases. Microorganisms use this process to break down the pectin in bark, releasing fibers in the process [8]. By adding particular enzymes to fibers, bioscouring is used to remove non-cellulosic contaminants. The enzymatic approach has replaced the harmful chemicals that were previously used in conventional scouring [9]. Other carbohydrases are added to pectinolytic enzymes to remove the sizing agent from cotton fibers without generating any negative side effects [10]. Similarly, pectinases are added to the process of tea and coffee fermentation in order to hydrolyze and break the cell wall of tea leaves and to remove the mucilage layer around the coffee beans, which reduces the ability of instant tea powder to froth. In contrast to the conventional technique, pectinase, xylanase, and cellulose from yeasts and bacteria improved the various black tea components [11].

For maximum yield of enzyme various fermentation factors are optimized. Previously one factor/variable at time technique was used for optimizing process parameters but it has many limitations like time consumption and laborious, superficial optimization, no factor interaction, and large number of experimental runs. Nowadays many statistical approaches are being practiced for optimization like Box–Behnken design, Plackett–Burman design (PBD), central composite design (CCD), and Graeco–Latin squares [12]. Among the statistical approaches, PBD method can quickly and efficiently filter out the most crucial parameters from a large number of factors using a single strategy. As a combination of different mathematical and statistical approaches or modeling, response surface methodology (RSM) is typically used to analyze the performance of complex systems and optimize the multiple process conditions [13]. This work aims to isolate and identify pectinase producing bacterial strains from indigenous soil samples and optimize fermentation conditions (both physical and chemical) to increase enzyme production initially by PBD and then followed by CCD.

2 Materials and methods

2.1 Isolation of pectinolytic bacteria

Soil samples were collected in winters from vegetable and fruit dump areas of district Haripur, Abbottabad and Mansehra. Isolation of collected strains was done by ten-fold serial dilution in Luria Basal (LB) media to reduce microbial load. From the last three dilutions, a small measured volume (100 µL) of each dilution was used to spread on LB plates containing yeast extract (10 g), sodium chloride (10 g), tryptone (10 g), agar (2 g), and 1% substrate. Initial pH of the medium was adjusted to 9 before sterilization at 121°C for 20 min. After inoculation with selected strain, the plates were incubated at 37°C for 24 h. After 24 h of incubation, separate distinct colonies were picked with sterilize wire loop and streaked on LB agar media plate and labeled accordingly. Then, the plates were again subjected to incubation at 37°C for 24 h for further growth. The process was repeated for a number of times to get the individual cell. Gram staining and biochemical assay were performed for initial identification of isolated species [14].

2.2 Qualitative screening

Bacterial strains were initially screened by inoculating in fermentation media containing glucose (2%), MgSO4 (0.05%), ammonium sulphate (0.3%), NaCl (0.1%), K2HpO4 (0.1%), and pectin substrate (1%). Selected colonies were inoculated on the media plates followed by incubation for 24 h at 37°C. After the complete incubation colonial plates were stained with 2% iodine dye, appearance of clear zone around colonies showed pectinolytic bacteria. The pectinolytic bacterial colonies were further qualitatively screened for optimum pectinase production on the basis of zone diameter around the colonies. Zone of hydrolysis was analyzed and enzyme index was calculated using the formula [15] as follows:

Enzyme index = Diameter of hydrolysis zone/Diameter of a colony.

2.3 Quantitative screening of selected bacterial strains

Submerged fermentation was used to evaluate the strains that showed pectinolytic activity in the qualitative assay. Fermentation media (100 mL) with 1% (v/v) inoculum was incubated in shaking incubator at 37°C for 24 h (agitation 150 rpm). After incubation period, the broth culture was centrifuged for 15 min at 8,000 rpm. The supernatant was used as crude enzyme for the enzyme assay. Then, 1 mL of substrate solution (1 g substrate in 100 mL distilled water) was mixed with 1 mL of cell free filtrate and incubated for 30 min in a water bath at 37°C. The reaction was then stopped with 3 mL of dinitrosalicylic acid reagent followed by boiling for 5 min. After cooling for 5 min in an ice bath, 720 µL distilled water was added to the reaction mixture and enzyme production was analyzed at 540 nm. Along with the test, a control was prepared by using heat-killed enzyme. All the experiments were performed in triplicate and the results are the mean values. According to standard assay, one unit of enzyme is defined as the quantity of enzyme required to release 1 mg of reducing sugar from galacturonic acid [16].

2.4 Phenotypic and biochemical characterizations

The highest pectinolytic isolates were characterized on the basis of various morphological, microscopic, and biochemical characteristics to identify the isolated strains to genus level as per results of Bergey’s Manual of Determinative Bacteriology. The test parameters were gram stain, catalase test, citrate, pectinase, lactose, oxidase, etc. Microscopic characteristics and colonial feature were also studied [17].

2.5 Molecular identification

Genomic DNA from maximum pectin producing strain was extracted according to the methods for bacterial DNA extraction by using Promega DNA extraction kit.

2.5.1 PCR of 16s rRNA gene of pectinolytic bacterial isolate

16s rRNA gene of highest pectinase producing strain was amplified by C-1000 Touch thermocycler using universal primer sequences. The PCR mixture was prepared by adding Hot start PCR master mix, forward primers (5′-AGA GTT TGA TCI TGG CTC AG-3′), reverse primers (5′-ACG GIT ACC TTG TTA CGA CTT-3′), and DNA template. The reaction was carried out at the conditions of denaturation at 94°C for 1 min followed by 35 cycles, 55°C annealing for 30 s, 72°C extension for 1 min and final elongation at 72°C for 10 min [18].

2.5.2 Gene sequencing and analysis of 16s rRNA

For gene sequencing, 16s rRNA sequence was analyzed by BioEdit Sequence Alignment Editor Software and compared to GenBank nucleotide data library. BLAST program of NCBI was used for the nucleotide sequence alignment and to determine its closest phylogenetic relatives. The 16S rRNA sequence was analyzed by MEGA11 software and compared to the sequences in GenBank nucleotide data library [19].

2.6 Statistical optimization of pectinase production by multi factorial experiments

The overall optimization of fermentative enzyme production was completed in three major steps. At first, various nutritional and physical parameters were evaluated for pectinase production by using PBD. In the next step, most significant variables were further optimized to their optimum level by using CCD. At last, the quality of model fitness was verified by computational analysis expressed by R 2, the coefficient of determination.

2.6.1 PBD

PBD was generated to assess the effect of the seven factors influencing pectinase enzyme production. Temperature, inoculum size, pH, incubation period, substrate concentration, metal ion, and nitrogen ion concentration were chosen as independent factors. All the selected variables were selected at low (−1) and high (+1) level (Table 1). Minitab software was used to create a design matrix of 12 experiments (Table 2). All experiments were performed in triplicate in 250 mL Erlenmeyer flasks containing 50 ml fermentation medium. PBD screening design depends on the first order model.

A = α 0 + ∑ α i B i .

Table 1

Variable levels screened in the PBD

Factor Low level (−1) High level (+1) Factor Low level (−1) High level (+1)
Temperature (°C) 25 45 Nitrogen source (%) 0.5 2.5
pH 6 10 Substrate (%) 1 5
Inoculum size (%) 1 5 Metal ion (%) Magnesium sulphate Sodium chloride
Incubation period (h) 24 96
Table 2

PBD for screening significant factors affecting pectinase production

Run B1: temp (°C) B2: pH B3: incubation period (h) B4: inoculum size (%) B5: metal ions B6: nitrogen source (%) B7: substrate (%) A: pectinase (IU/mL)
1 45 6 96 1 MgSO4 0.5 5 0.43
2 45 10 24 5 MgSO4 0.5 1 0.21
3 25 10 96 1 NaCl 0.5 1 0.43
4 45 6 96 5 MgSO4 2.5 1 0.59
5 45 10 24 5 NaCl 0.5 5 0.43
6 45 10 96 1 NaCl 2.5 1 0.59
7 25 10 96 5 MgSO4 2.5 5 0.3
8 25 6 96 5 MgSO4 0.5 5 0.42
9 25 6 24 5 MgSO4 2.5 1 0.59
10 45 6 24 1 MgSO4 2.5 5 0.83
11 25 10 24 1 MgSO4 2.5 5 0.3
12 25 6 24 1 MgSO4 0.5 1 0.35

Here A represents the response, i.e., pectinase activity, α 0 is the model intercept, α i is the variable estimate, and B i shows the independent variable levels. Significance of each variable was best demonstrated by Pareto Plot. All the variables with p-values less than 0.05 were noted as significant. Statistical significance of the model was also determined by T and R 2 values. All factors that were found statistically significant were chosen for further steps [20].

2.6.2 Optimization of significant variables using CCD

CCD of RSM was employed to optimize significant factors for pectinase activity. Number of experimental combinations in CCD is 2 h + 2 h + no, where h represents the number of independent variables and no represents the repetition of experiments at center point. Four factors were identified as significant during the initial screening of factors (in PBD), including initial temperature, pH, nitrogen source, and NaCl were further optimized using CCD in RSM. Three levels for each variable are low (−),central (0), and high (+) (Table 3). A second-order polynomial equation was used to analyze the effect of these variables on response.

A = α 0 + Σ α i B i + Σ α i i B i 2 + Σ α i j B i B j ,

where A = predicted response, α0 = intercept term, α i = linear effect, α ii = squared effect, α ij = interaction effect, and B i and B j = independent variables. An experimental model of 26 runs was designed (Table 4). Pectinase activity was determined and used as a response, while the rest of the optimized factors were kept constant for subsequent designed experimental runs [21].

Table 3

Factors with their levels in CCD

Factors Levels
−1 0 +1
NaCl concentration (%) 0.05 0.075 0.08
pH 6 7 8
Nitrogen source concentration (%) 0.5 1.5 1.7
Temperature 33°C 37°C 45°C
Table 4

CCD Optimization of pectinase production (IU/mL)

Run order NaCl pH Nitrogen source Temperature °C IU/mL
1 0.07 7.6 1.3 33 1.95
2 0.08 7.6 1.3 33 5.4
3 0.07 8.4 1.3 33 4.8
4 0.08 8.4 1.3 33 9
5 0.07 7.6 1.7 33 9
6 0.08 7.6 1.7 33 9.6
7 0.07 8.4 1.7 33 11.7
8 0.08 8.4 1.7 33 8.7
9 0.07 7.6 1.3 37 9
10 0.08 7.6 1.3 37 10.2
11 0.07 8.4 1.3 37 9.6
12 0.08 8.4 1.3 37 5.7
13 0.07 7.6 1.7 37 15.3
14 0.08 7.6 1.7 37 1.2
15 0.07 8.4 1.7 37 19.2
16 0.08 8.4 1.7 37 3.3
17 0.075 8 1.5 35 10.8
18 0.075 8 1.5 35 4.95
19 0.05 8 1.5 35 14.4
20 0.1 8 1.5 35 4.5
21 0.075 6 1.5 35 6.75
22 0.075 10 1.5 35 13.6
23 0.075 8 0.5 35 2.1
24 0.075 8 2.5 35 14.4
25 0.075 8 1.5 25 6.6
26 0.075 8 1.5 45 9.6

2.7 Model validation

Designed model in the RSM was validated by different point studies. Obtained data were compared to predicted values and prediction errors.

2.8 Statistical analysis of data

All the designed run orders were carried out in triplicate and response was expressed as mean of observed values. ANOVA analysis was applied to the model data and the quality of polynomial equation in term of fit was accessed by evaluating the R 2 coefficient to Adj R 2 coefficient; model significance was checked by F test and P value. 3D surface and 2D contour plots were plotted to express the significance of independent variables on the results.

3 Results

3.1 Screening for maximum pectinase producing bacteria

Strains with particularly considering their conditions were isolated from soil samples and were qualitatively analyzed for pectinase production by dropping iodine dye on pectinase agar plate. Eight strains (SH1, SH3, SH5, SH7, SH16, SH17, SH18, and SH25) showed positive results with the formation of clear zone surrounding the colonies due to pectin hydrolysis at dark purple background. The largest zone was observed around strain SH7 (Figure 1). Quantitative screening of the potent pectinolytic isolates was conducted using the enzyme activity assay. The highest pectinolytic activity was shown by isolate SH7 (0.77 IU/mL) which was consistent with the results obtained from the previous qualitative assessment. Due to its potent activity, SH7 strain was chosen for further investigation (Figure 2).

Figure 1 (a) Pectinase producers and (b) non-producers.
Figure 1

(a) Pectinase producers and (b) non-producers.

Figure 2 Qualitative screening of pectinase enzyme.
Figure 2

Qualitative screening of pectinase enzyme.

3.2 Characterization of pectinolytic bacterial isolate

Highest pectinase producing isolate, i.e., SH7 strain was subjected to morphological, microscopic, and biochemical characterization according to methods of Bergey’s Manual of Determinative Bacteriology. The results showed that isolate SH7 belongs to the genus Bacillus, gram (+), citrate (+), catalase (+), oxidase (−), lactose (+), glucose (+), urease (+), pectinase (+), short rod shaped in chains, aerobic, and spore forming organism.

3.3 Molecular identification of pectinolytic bacterial isolate (SH7)

SH7 strain was subjected to molecular identification for further confirmation and identification. The PCR was used to amplify 16s rRNA of Bacillus SH7. Through agarose gel electrophoresis band of 1.5 kb was obtained. After purification and sequencing of reamplified 16s rRNA, 1,400 nucleotide sequence was obtained that contained both variable and conserved sections. The sequenced data were run to the BLAST study for identification of the bacterial strain (SH7). Phylogenetic tree was constructed on the basis of 16s rRNA sequences of pectinase producing SH7 strain and close species sequences were downloaded from NCBI. The sequences were aligned by CLUSTAL W2 and neighbor-joining tree was constructed in MEGA11 by Kimura-2-parameter [22]. Phylogenetic tree from Bacillus thuringiensis showed 100% similarity with their closest phylogenetic relatives Bacillus thuringiensis LAA3 in the NCBI (Figure 3).

Figure 3 Electrophoresis shows PCR products of 16s rRNA of Bacillus thuringiensis SH7 (M) (a) and phylogenetic tree using multiple CLUSTAL W2 alignment software of MEGA 11 (b).
Figure 3

Electrophoresis shows PCR products of 16s rRNA of Bacillus thuringiensis SH7 (M) (a) and phylogenetic tree using multiple CLUSTAL W2 alignment software of MEGA 11 (b).

3.4 Multi factorial optimization designs for pectinase production

Current study is focused on statistical approach to gradually enhance pectinase production by using different as well as minimize cost.

3.4.1 PBD

PBD was conducted to find out the significant variables that affect the production of pectinase. Totally 12 experiments were designed at 2 levels i.e., high and low. Temperature, incubation period, pH, incubation temperature, nitrogen source, metal ions, and substrate concentration were primarily selected as optimization parameters. Table 2 demonstrates the average pectinase activity (IU/mL). The lowest pectinase activity (0.2 IU/mL) was observed in run order 2 whereas highest pectinase activity (0.8 IU/mL) was obtained in run over 10 having pH 6, sodium chloride as metal ion, incubation period of 24 h, inoculum size of 1%, nitrogen source of 2.5%, substrate concentration of 5%, and temperature of 450°C. Multi regression analysis on the experimental model established the first order polynomial equation to show the production of pectinase as follows:

A ( IU/mL ) = 0.503 + 0.00575 B 1 − 0.03958 B 2 + 0.000116 B 3 – 0.01625 B 4 + 0.0925 B 5 + 0.0775 B 6 − 0.00208 B 7 ,

where A is the pectinase activity; B1 is the temperature; B2 is the pH; B3 is the incubation period; B4 is the inoculum size; B5 is the metal ions; B6 is the nitrogen source; and B7 is the substrate concentration. Regression analysis in the form of Pareto chart showed that four factors including temperature (P = 0.031), pH (P = 0.011), nitrogen source (P = 0.012), and metal ions (P = 0.006) have significant effects on pectinase production because these factors clearly crossed the reference line (Figure 4).

Figure 4 Pareto chart (PBD) showing effect of different factors on pectinase production.
Figure 4

Pareto chart (PBD) showing effect of different factors on pectinase production.

3.4.2 CCD

Second order model (CCD) was performed to analyze the four factors (pH, temperature, nitrogen source, and metal ions) that were found as significant factors in PBD while making non-significant factors constant as per highest run over values. The overall model summary showed that the highest pectinase activity (19.2 IU/mL) was observed in run over 15 whereas lowest pectinase activity of 1.2 IU/mL was observed in run over 14. Regression analysis for the experimental data showed that predicted response can be obtained by using second-order polynomial equation as follows:

A ( IU/mL ) = − 1 , 141 + 1 , 3181 B 1 + 18.5 B 2 + 215 B 3 + 23.5 B 4 + 5 , 511 B 1 ⁎ B 1 + 1.04 B 2 ⁎ B 2 + 2.2 B 3 ⁎ B 3 + 0.021 B 4 ⁎ B 4 − 305 B 1 ⁎ B 2 – 2 , 334 B 1 ⁎ B 3 − 237.2 B 1 ⁎ B 4 + 4.10 B 2 ⁎ B 3 − 0.480 B 2 ⁎ B 4 − 2.09 B 3 ⁎ B 4 ,

where A is the pectinase activity; B1 is sodium chloride; B2 is the pH; B3 is the nitrogen source; and B4 is the temperature. ANOVA analysis for pectinase production was employed on CCD design (Table 5). High precision of model was showed by low magnitude of P value (at 1% level of significance). Significance of each coefficient was indicated as lower P and higher T values. Sodium chloride (P = 0.004), pH (0.055), and nitrogen source (0.002) were seen as significant linear term whereas no square term was found significant throughout the design. Two-way interaction showed significant value for sodium chloride and nitrogen (0.004), and sodium chloride and temperature (0.004) interactions. Significance of model was also depicted by values of R 2 (coefficient of determination). In the current experimental model, value of R 2 (87.47%) was found high as compare to adjusted R 2 (68.67%) that also confirms the accuracy of designed experiments.

Table 5

Variance analysis of CCD for pectinase production

Source DF Seq SS Contribution (%) Adj SS Adj MS F-Value P Value
Model 15 445.912 87.47 445.912 29.727 4.65
Blocks 1 2.666 0.52 0.214 0.214 0.03
Linear 4 238.561 46.79 238.561 59.640 9.33
B1 1 89.717 17.60 89.717 89.717 14.04 0.004
B2 1 30.139 5.91 30.139 30.139 4.72 0.055
B3 1 106.528 20.90 106.528 106.528 16.67 0.002
B4 1 12.178 2.39 12.178 12.178 1.91 0.197
Square 4 6.317 1.24 6.317 1.579 0.25
B1*B1 1 0.596 0.12 0.531 0.531 0.08 0.779
B2*B2 1 5.486 1.08 0.778 0.778 0.12 0.734
B3*B3 1 0.039 0.01 0.225 0.225 0.04 0.855
B4*B4 1 0.196 0.196 0.196 0.196 0.03 0.864
Two-way interaction 6 198.368 38.91 198.368 33.061 5.17
B1*B2 1 5.941 1.17 5.941 5.941 0.93 0.358
B1*B3 1 87.189 17.10 87.189 87.189 13.65 0.004
B1*B4 1 90.013 17.66 90.013 90.013 14.09 0.004
B2*B3 1 1.723 0.34 1.723 1.723 0.27 0.615
B2*B4 1 2.364 0.46 2.364 2.364 0.37 0.557
B3*B4 1 11.139 2.18 11.139 11.139 1.74 0.216
Error 10 63.891 12.53 63.891 6.389
Lack-of-fit 9 46.779 9.18 46.779 5.198 0.30
Pure error 1 17.111 3.36 17.111 17.111
Total 25 509.803 100.00

B1 = Sodium chloride, B2 = pH, B3 = Nitrogen source, B4 = Temperature.

The response analysis was obtained after running the CCD design. Pareto charts were plotted to determine the significant effects for pectinase enzyme. In the Pareto chart, three factors including metal ion (NaCl) (P value 0.004), pH (P value 0.05), and nitrogen source (P value 0.002) were found statistically significant, as these factors clearly crossed the reference line (Figure 5).

Figure 5 Pareto chart showing the effect of NaCl, pH, nitrogen source, and temperature on pectinase by B. thuringiensisstrain-SH7.
Figure 5

Pareto chart showing the effect of NaCl, pH, nitrogen source, and temperature on pectinase by B. thuringiensisstrain-SH7.

Expression of model regression equation in the form of surface plots (3D) and contour plots (2D) graphs provides a rapid and structured way to visualize the best values of significant variable. Elliptical contours show maximum interaction among the selected variables whereas circular shaped contour explains no interaction among variables. 3D surface plots further explain the relationship between dependent and independent variables. As depicted by these graphical representations, nitrogen concentration of 0.657386, sodium chloride of 0.075, and pH 8 resulted in a remarkable increase in the enzyme yield (22.5 IU/mL) as compared to the results of PBD (0.8 IU/mL) (Figure 6).

Figure 6 Response surface 2D and 3D plots representing interaction between variables affecting pectinase production: (a) pH and Nacl, (b) Nacl and nitrogen source, (c) temperature and Nacl, (d) nitrogen and pH, (e) temperature and pH, and (f) temperature and nitrogen source.
Figure 6 Response surface 2D and 3D plots representing interaction between variables affecting pectinase production: (a) pH and Nacl, (b) Nacl and nitrogen source, (c) temperature and Nacl, (d) nitrogen and pH, (e) temperature and pH, and (f) temperature and nitrogen source.
Figure 6

Response surface 2D and 3D plots representing interaction between variables affecting pectinase production: (a) pH and Nacl, (b) Nacl and nitrogen source, (c) temperature and Nacl, (d) nitrogen and pH, (e) temperature and pH, and (f) temperature and nitrogen source.

4 Discussion

Nowadays chemicals have been replaced with biomolecules for accelerating various reactions. Among these biomolecules, enzymes are most commonly used in many industrial sectors. It is important to synthesize these biocatalysts with enhanced and desired characteristics that can meet the industrial demands. Pectinases are among industrially important enzymes that have numerous applications in various fields. Although, pectinases are broadly synthesized by higher plants but advances in microbial biotechnology have revealed the enormous pectinolytic potential of microbes, particularly bacteria and filamentous fungi. In general, filamentous fungi are the basis of acidic pectinases, whereas bacteria are the source of alkaline pectinases [23]. Bacteria have various advantages over filamentous fungi as they do not experience rheological issues in fermentation broth and do not produce a mixture of pectinases [24]. Locally isolated bacteria are yet to be investigated for biotechnological applications [25].

The current study was conducted with the isolation of 50 microbial isolates from soil of fruit and vegetable dumps areas. Indeed soil dumped with rotten vegetables and fruits is a well-known productive habitat in order to isolate pectinolytic bacteria because pectin promotes their growth by supplying nutritious requirements [26]. The qualitative screening of bacterial isolates was carried out by growing them on pectin fermentation media and treating with potassium iodide solution. Potassium iodide is useful because when it is oxidized, a color change is produced which shows the presence of pectin polymer that gives color change [27]. Following the initial screening, 5 out of 50 (10%) strains were obtained as potential pectinase producer. Our study is in correspondence with the findings of Merín et al. [19]. Quantitative enzyme production showed that SH7 has maximum enzyme production ability as compared to other strains. SH7 strain was chosen for further study and subjected to colonial, microscopic, biochemical characterization, and molecular identification. The results showed that selected bacterial isolate displayed 100% similarity with other reported strains of Bacillus thuringiensis. Liu and coworkers also reported identification of pectinase producing strain as Bacillus thuringiensis [28]. Similarly, Namasivayam isolated bacterial strain from soil sample and identified it as Bacillus thuringiensis [29].

Generally, the production of pectinase is majorly affected by biological, physical, and chemical parameters. The production of enzymes from microbes can be improved by analyzing the influence of nutritional and ecological factors mostly known as optimization methods which ultimately results in decrease in production cost. Conventionally, factors that affect the production process were studied one after another by keeping the other factors constant. In this strategy, the influence of values and factors on the analysis can be obtained separately. However, this technique is found as laborious and does not explain the interactions between variables [30]. Later, different statistical techniques have been developed for the optimization of biological and chemical factors. In the current research, two levels of statistical designs were used in order to perform optimization, the first one is PBD and the second is CCD.

PBD is an effective method for screening the influence of several factors on the response [31]. 12 runs of the experiment were performed as obtained by PBD. Maximum pectinase units 0.83 IU/mL was produced in PBD when fermentation was carried for 24 h at 450°C by maintaining the medium at pH 10, inoculum size of 5%, substrate concentration of 1%, nitrogen source of 0.5%, and MnSO4 as metal ion. The results of recent work are in accordance with the findings of Ahmed et al. [5]. Factorial regression analysis of PBD design was performed that showed significance of multiple factors. Among all test variables, temperature, pH, nitrogen source, and metal ion concentration were statistically significant for pectinase production as these factors have positive R-coefficient and P value less than 0.05. In the current model, value of R 2 (87.47%) was found high as compared to adjusted R 2 (68.67%) that also confirms the accuracy of designed experiments. Accuracy of optimization design for pectinase production in terms of R 2 value was also reported in an earlier study (99.93%) [32].

Pareto chart clearly shows significant effects for enzyme by a reference line. Exceeding bars from reference line shows significant factors for response analysis. In a recent study, response analysis was obtained after running the PBD design. The most significant factors for production of pectinase were sodium chloride, temperature, pH, and nitrogen source. Our results are found in accordance with the findings of Venil et al., who reported pH and nitrogen source as significant factor by response analysis in terms of Pareto [33].

During statistical analysis, PBD was used to screen significant factors that have profound effect on enzyme production. In the current study, the significant factors by PBD design were further optimized by using central composite. In CCD, the overall model summary showed that the significant factors are pH, nitrogen source, and metal ion concentrations that showed the maximum pectinase yield of (22.5 IU/mL) at 0.657% nitrogen source, 0.075% NaCl, and pH 8. Accuracy of CCD design was also checked by using ANOVA analysis. Each significant coefficient was analyzed by lower P and higher T values. All factors showing p values less than 0.05 was a significant variable. Whereas no square term was found significant throughout the design. A two-way interaction showed significant value for sodium chloride and nitrogen (0.004) and sodium chloride, and temperature (0.004) interactions. Our results are in contrast with earlier findings that showed maximum pectinase production (10 IU/mL) increase in the pectinase activity at 40°C and 24 h by Aspergillus niger [34]. In the current experimental model, the value of R 2 (87.47%) was found high as compared to adjusted R 2 (68.67%) that also confirms the accuracy of designed experiments [35].

Representation of the model regression equation in the form of surface plots (3D) and contour plots (2D) graphs provides a rapid and structured way to visualize the best values of significant variable. Multiple combinations among two variables (keeping all the other selected parameters at their respective zero level) are graphically presented in the form of contour curves. Elliptical contours show maximum interaction among selected variables whereas circular shaped contour explains no interaction among variables [36]. 3D surface plots further explain the relationship between independent and dependent variables. As depicted by these graphical representations, nitrogen concentration of 0.657386, sodium chloride of 0.075, and pH 8 resulted in a significant increase in the enzyme yield (22.5 IU/mL) as compared to the results of PBD (0.8 IU/mL). Significant interaction of fermentation variables such as CaCl2 (0.8%), temperature (40°C), inoculum size (1.5%), and pectin concentration (2.5 g/L) for optimum pectinase yield by B. subtilis BKDS1 was also reported in previous findings [37].

The current study resulted in isolation of pectinolytic bacterial strain from indigenous soil samples. Moreover, optimization studies showed that significant titer of enzyme could be used for different industrial applications.

5 Conclusion

Members of genus bacillus have been reported for production of biotechnologically important products. Among these commercially important products, enzymes are well documented by Bacillus thuringiensis. The current study was focused on isolation of pectinolytic bacillus strain from soil sample. Optimum pectinase producing strain was identified as Bacillus thuringiensis SH7 on the basis of microscopic, morphological, biochemical, and molecular characteristics. Pectinase production was optimized in two-step process, PBD followed by CCD. Optimization of fermentation variable by PBD showed that among seven selected variables, temperature, sodium chloride, nitrogen source, and pH have a remarkable effect on extracellular pectinase production. The significant factors by PBD were optimized to their optimal level by CCD of RSM. Temperature, NaCl, and pH were also found as optimum variables after CCD analysis. The overall optimization model was found significant in terms of R, P, and F values. The present study resulted in a significant statistical optimization of enzyme that can be used for different biotechnologically important processes.

Acknowledgements

Authors are thankful to the Researchers Supporting Project number (RSP2023R197), King Saud University, Riyadh, Saudi Arabia.

  1. Funding information: This research was funded by Researchers Supporting Project number (RSP2023R197), King Saud University, Riyadh, Saudi Arabia.

  2. Author contributions: Conceptualization: I.K.; methodology: I.K.; software: M.U.R.; validation: A.H.; formal analysis: A.R.; investigation: K.A.; resources: M.U.R.; data curation: K.A.; writing – review and editing: I.K.; visualization: M.K., A.A., and M.A.; supervision: I.K.; project administration: M.U.R.; and funding acquisition: A.A. and M.A. All authors have read and agreed to the published version of the manuscript.

  3. Conflict of interest: The authors declare no conflict of interest.

  4. Ethical approval: The conducted research is not related to either human or animal use.

  5. Data availability statement: The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.

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Received: 2023-04-19
Revised: 2023-06-02
Accepted: 2023-06-22
Published Online: 2023-07-24

© 2023 the author(s), published by De Gruyter

This work is licensed under the Creative Commons Attribution 4.0 International License.

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  142. A comprehensive review of non-alkaloidal metabolites from the subfamily Amaryllidoideae (Amaryllidaceae)
  143. Discovery of the chemical constituents, structural characteristics, and pharmacological functions of Chinese caterpillar fungus
  144. Eco-friendly green approach of nickel oxide nanoparticles for biomedical applications
  145. Advances in the pharmaceutical research of curcumin for oral administration
  146. Rapid Communication
  147. Determination of the contents of bioactive compounds in St. John’s wort (Hypericum perforatum): Comparison of commercial and wild samples
  148. Retraction
  149. Retraction of “Two mixed-ligand coordination polymers based on 2,5-thiophenedicarboxylic acid and flexible N-donor ligands: The protective effect on periodontitis via reducing the release of IL-1β and TNF-α”
  150. Topical Issue on Phytochemicals, biological and toxicological analysis of aromatic medicinal plants
  151. Anti-plasmodial potential of selected medicinal plants and a compound Atropine isolated from Eucalyptus obliqua
  152. Anthocyanin extract from black rice attenuates chronic inflammation in DSS-induced colitis mouse model by modulating the gut microbiota
  153. Evaluation of antibiofilm and cytotoxicity effect of Rumex vesicarius methanol extract
  154. Chemical compositions of Litsea umbellata and inhibition activities
  155. Green synthesis, characterization of silver nanoparticles using Rhynchosia capitata leaf extract and their biological activities
  156. GC-MS analysis and antibacterial activities of some plants belonging to the genus Euphorbia on selected bacterial isolates
  157. The abrogative effect of propolis on acrylamide-induced toxicity in male albino rats: Histological study
  158. A phytoconstituent 6-aminoflavone ameliorates lipopolysaccharide-induced oxidative stress mediated synapse and memory dysfunction via p-Akt/NF-kB pathway in albino mice
  159. Anti-diabetic potentials of Sorbaria tomentosa Lindl. Rehder: Phytochemistry (GC-MS analysis), α-amylase, α-glucosidase inhibitory, in vivo hypoglycemic, and biochemical analysis
  160. Assessment of cytotoxic and apoptotic activities of the Cassia angustifolia aqueous extract against SW480 colon cancer
  161. Biochemical analysis, antioxidant, and antibacterial efficacy of the bee propolis extract (Hymenoptera: Apis mellifera) against Staphylococcus aureus-induced infection in BALB/c mice: In vitro and in vivo study
  162. Assessment of essential elements and heavy metals in Saudi Arabian rice samples underwent various processing methods
  163. Two new compounds from leaves of Capparis dongvanensis (Sy, B. H. Quang & D. V. Hai) and inhibition activities
  164. Hydroxyquinoline sulfanilamide ameliorates STZ-induced hyperglycemia-mediated amyleoid beta burden and memory impairment in adult mice
  165. An automated reading of semi-quantitative hemagglutination results in microplates: Micro-assay for plant lectins
  166. Inductively coupled plasma mass spectrometry assessment of essential and toxic trace elements in traditional spices consumed by the population of the Middle Eastern region in their recipes
  167. Phytochemical analysis and anticancer activity of the Pithecellobium dulce seed extract in colorectal cancer cells
  168. Impact of climatic disturbances on the chemical compositions and metabolites of Salvia officinalis
  169. Physicochemical characterization, antioxidant and antifungal activities of essential oils of Urginea maritima and Allium sativum
  170. Phytochemical analysis and antifungal efficiency of Origanum majorana extracts against some phytopathogenic fungi causing tomato damping-off diseases
  171. Special Issue on 4th IC3PE
  172. Graphene quantum dots: A comprehensive overview
  173. Studies on the intercalation of calcium–aluminium layered double hydroxide-MCPA and its controlled release mechanism as a potential green herbicide
  174. Synergetic effect of adsorption and photocatalysis by zinc ferrite-anchored graphitic carbon nitride nanosheet for the removal of ciprofloxacin under visible light irradiation
  175. Exploring anticancer activity of the Indonesian guava leaf (Psidium guajava L.) fraction on various human cancer cell lines in an in vitro cell-based approach
  176. The comparison of gold extraction methods from the rock using thiourea and thiosulfate
  177. Special Issue on Marine environmental sciences and significance of the multidisciplinary approaches
  178. Sorption of alkylphenols and estrogens on microplastics in marine conditions
  179. Cytotoxic ketosteroids from the Red Sea soft coral Dendronephthya sp.
  180. Antibacterial and biofilm prevention metabolites from Acanthophora spicifera
  181. Characteristics, source, and health risk assessment of aerosol polyaromatic hydrocarbons in the rural and urban regions of western Saudi Arabia
  182. Special Issue on Advanced Nanomaterials for Energy, Environmental and Biological Applications - Part II
  183. Green synthesis, characterization, and evaluation of antibacterial activities of cobalt nanoparticles produced by marine fungal species Periconia prolifica
  184. Combustion-mediated sol–gel preparation of cobalt-doped ZnO nanohybrids for the degradation of acid red and antibacterial performance
  185. Perinatal supplementation with selenium nanoparticles modified with ascorbic acid improves hepatotoxicity in rat gestational diabetes
  186. Evaluation and chemical characterization of bioactive secondary metabolites from endophytic fungi associated with the ethnomedicinal plant Bergenia ciliata
  187. Enhancing photovoltaic efficiency with SQI-Br and SQI-I sensitizers: A comparative analysis
  188. Nanostructured p-PbS/p-CuO sulfide/oxide bilayer heterojunction as a promising photoelectrode for hydrogen gas generation
https://doi.org/10.1515/chem-2022-0358
Schlagwörter für diesen Artikel
Plackett–Burman design; central composite design; pectinases; optimization; fermentation
Creative Commons
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Artikel in diesem Heft

  1. Characteristics, source, and health risk assessment of aerosol polyaromatic hydrocarbons in the rural and urban regions of western Saudi Arabia
  2. Regular Articles
  3. A network-based correlation research between element electronegativity and node importance
  4. Pomegranate attenuates kidney injury in cyclosporine-induced nephrotoxicity in rats by suppressing oxidative stress
  5. Ab initio study of fundamental properties of XInO3 (X = K, Rb, Cs) perovskites
  6. Responses of feldspathic sandstone and sand-reconstituted soil C and N to freeze–thaw cycles
  7. Robust fractional control based on high gain observers design (RNFC) for a Spirulina maxima culture interfaced with an advanced oxidation process
  8. Study on arsenic speciation and redistribution mechanism in Lonicera japonica plants via synchrotron techniques
  9. Optimization of machining Nilo 36 superalloy parameters in turning operation
  10. Vacuum impregnation pre-treatment: A novel method for incorporating mono- and divalent cations into potato strips to reduce the acrylamide formation in French fries
  11. Characterization of effective constituents in Acanthopanax senticosus fruit for blood deficiency syndrome based on the chinmedomics strategy
  12. Comparative analysis of the metabolites in Pinellia ternata from two producing regions using ultra-high-performance liquid chromatography–electrospray ionization–tandem mass spectrometry
  13. The assessment of environmental parameter along the desalination plants in the Kingdom of Saudi Arabia
  14. Effects of harpin and carbendazim on antioxidant accumulation in young jujube leaves
  15. The effects of in ovo injected with sodium borate on hatching performance and small intestine morphology in broiler chicks
  16. Optimization of cutting forces and surface roughness via ANOVA and grey relational analysis in machining of In718
  17. Essential oils of Origanum compactum Benth: Chemical characterization, in vitro, in silico, antioxidant, and antibacterial activities
  18. Translocation of tungsten(vi) oxide/gadolinium(iii) fluoride in tellurite glasses towards improvement of gamma-ray attenuation features in high-density glass shields
  19. Mechanical properties, elastic moduli, and gamma ray attenuation competencies of some TeO2–WO3–GdF3 glasses: Tailoring WO3–GdF3 substitution toward optimum behavioral state range
  20. Comparison between the CIDR or sponge with hormone injection to induce estrus synchronization for twining and sex preselection in Naimi sheep
  21. Exergetic performance analyses of three different cogeneration plants
  22. Psoralea corylifolia (babchi) seeds enhance proliferation of normal human cultured melanocytes: GC–MS profiling and biological investigation
  23. A novel electrochemical micro-titration method for quantitative evaluation of the DPPH free radical scavenging capacity of caffeic acid
  24. Comparative study between supported bimetallic catalysts for nitrate remediation in water
  25. Persicaline, an alkaloid from Salvadora persica, inhibits proliferation and induces apoptosis and cell-cycle arrest in MCF-7 cells
  26. Determination of nicotine content in locally produced smokeless tobacco (Shammah) samples from Jazan region of Saudi Arabia using a convenient HPLC-MS/MS method
  27. Changes in oxidative stress markers in pediatric burn injury over a 1-week period
  28. Integrated geophysical techniques applied for petroleum basins structural characterization in the central part of the Western Desert, Egypt
  29. The impact of chemical modifications on gamma-ray attenuation properties of some WO3-reinforced tellurite glasses
  30. Microwave and Cs+-assisted chemo selective reaction protocol for synthesizing 2-styryl quinoline biorelevant molecules
  31. Structural, physical, and radiation absorption properties of a significant nuclear power plant component: A comparison between REX-734 and 316L SS austenitic stainless steels
  32. Effect of Moringa oleifera on serum YKL-40 level: In vivo rat periodontitis model
  33. Investigating the impact of CO2 emissions on the COVID-19 pandemic by generalized linear mixed model approach with inverse Gaussian and gamma distributions
  34. Influence of WO3 content on gamma rays attenuation characteristics of phosphate glasses at low energy range
  35. Study on CO2 absorption performance of ternary DES formed based on DEA as promoting factor
  36. Performance analyses of detonation engine cogeneration cycles
  37. Sterols from Centaurea pumilio L. with cell proliferative activity: In vitro and in silico studies
  38. Untargeted metabolomics revealing changes in aroma substances in flue-cured tobacco
  39. Effect of pumpkin enriched with calcium lactate on iron status in an animal model of postmenopausal osteoporosis
  40. Energy consumption, mechanical and metallographic properties of cryogenically treated tool steels
  41. Optimization of ultra-high pressure-assisted extraction of total phenols from Eucommia ulmoides leaves by response surface methodology
  42. Harpin enhances antioxidant nutrient accumulation and decreases enzymatic browning in stored soybean sprouts
  43. Physicochemical and biological properties of carvacrol
  44. Radix puerariae in the treatment of diabetic nephropathy: A network pharmacology analysis and experimental validation
  45. Anti-Alzheimer, antioxidants, glucose-6-phosphate dehydrogenase effects of Taverniera glabra mediated ZnO and Fe2O3 nanoparticles in alloxan-induced diabetic rats
  46. Experimental study on photocatalytic CO2 reduction performance of ZnS/CdS-TiO2 nanotube array thin films
  47. Epoxy-reinforced heavy metal oxides for gamma ray shielding purposes
  48. Black mulberry (Morus nigra L.) fruits: As a medicinal plant rich in human health-promoting compounds
  49. Promising antioxidant and antimicrobial effects of essential oils extracted from fruits of Juniperus thurifera: In vitro and in silico investigations
  50. Chloramine-T-induced oxidation of Rizatriptan Benzoate: An integral chemical and spectroscopic study of products, mechanisms and kinetics
  51. Study on antioxidant and antimicrobial potential of chemically profiled essential oils extracted from Juniperus phoenicea (L.) by use of in vitro and in silico approaches
  52. Screening and characterization of fungal taxol-producing endophytic fungi for evaluation of antimicrobial and anticancer activities
  53. Mineral composition, principal polyphenolic components, and evaluation of the anti-inflammatory, analgesic, and antioxidant properties of Cytisus villosus Pourr leaf extracts
  54. In vitro antiproliferative efficacy of Annona muricata seed and fruit extracts on several cancer cell lines
  55. An experimental study for chemical characterization of artificial anterior cruciate ligament with coated chitosan as biomaterial
  56. Prevalence of residual risks of the transfusion-transmitted infections in Riyadh hospitals: A two-year retrospective study
  57. Computational and experimental investigation of antibacterial and antifungal properties of Nicotiana tabacum extracts
  58. Reinforcement of cementitious mortars with hemp fibers and shives
  59. X-ray shielding properties of bismuth-borate glass doped with rare earth ions
  60. Green supported silver nanoparticles over modified reduced graphene oxide: Investigation of its antioxidant and anti-ovarian cancer effects
  61. Orthogonal synthesis of a versatile building block for dual functionalization of targeting vectors
  62. Thymbra spicata leaf extract driven biogenic synthesis of Au/Fe3O4 nanocomposite and its bio-application in the treatment of different types of leukemia
  63. The role of Ag2O incorporation in nuclear radiation shielding behaviors of the Li2O–Pb3O4–SiO2 glass system: A multi-step characterization study
  64. A stimuli-responsive in situ spray hydrogel co-loaded with naringenin and gentamicin for chronic wounds
  65. Assessment of the impact of γ-irradiation on the piperine content and microbial quality of black pepper
  66. Antioxidant, sensory, and functional properties of low-alcoholic IPA beer with Pinus sylvestris L. shoots addition fermented using unconventional yeast
  67. Screening and optimization of extracellular pectinase produced by Bacillus thuringiensis SH7
  68. Determination of polyphenols in Chinese jujube using ultra-performance liquid chromatography–mass spectrometry
  69. Synergistic effects of harpin and NaCl in determining soybean sprout quality under non-sterile conditions
  70. Field evaluation of different eco-friendly alternative control methods against Panonychus citri [Acari: Tetranychidae] spider mite and its predators in citrus orchards
  71. Exploring the antimicrobial potential of biologically synthesized zero valent iron nanoparticles
  72. NaCl regulates goldfish growth and survival at three food supply levels under hypoxia
  73. An exploration of the physical, optical, mechanical, and radiation shielding properties of PbO–MgO–ZnO–B2O3 glasses
  74. A novel statistical modeling of air pollution and the COVID-19 pandemic mortality data by Poisson, geometric, and negative binomial regression models with fixed and random effects
  75. Treatment activity of the injectable hydrogels loaded with dexamethasone In(iii) complex on glioma by inhibiting the VEGF signaling pathway
  76. An alternative approach for the excess lifetime cancer risk and prediction of radiological parameters
  77. Panax ginseng leaf aqueous extract mediated green synthesis of AgNPs under ultrasound condition and investigation of its anti-lung adenocarcinoma effects
  78. Study of hydrolysis and production of instant ginger (Zingiber officinale) tea
  79. Novel green synthesis of zinc oxide nanoparticles using Salvia rosmarinus extract for treatment of human lung cancer
  80. Evaluation of second trimester plasma lipoxin A4, VEGFR-1, IL-6, and TNF-α levels in pregnant women with gestational diabetes mellitus
  81. Antidiabetic, antioxidant and cytotoxicity activities of ortho- and para-substituted Schiff bases derived from metformin hydrochloride: Validation by molecular docking and in silico ADME studies
  82. Antioxidant, antidiabetic, antiglaucoma, and anticholinergic effects of Tayfi grape (Vitis vinifera): A phytochemical screening by LC-MS/MS analysis
  83. Identification of genetic polymorphisms in the stearoyl CoA desaturase gene and its association with milk quality traits in Najdi sheep
  84. Cold-acclimation effect on cadmium absorption and biosynthesis of polyphenolics, and free proline and photosynthetic pigments in Spirogyra aequinoctialis
  85. Analysis of secondary metabolites in Xinjiang Morus nigra leaves using different extraction methods with UPLC-Q/TOF-MS/MS technology
  86. Nanoarchitectonics and performance evaluation of a Fe3O4-stabilized Pickering emulsion-type differential pressure plugging agent
  87. Investigating pyrolysis characteristics of Shengdong coal through Py-GC/MS
  88. Extraction, phytochemical characterization, and antifungal activity of Salvia rosmarinus extract
  89. Introducing a novel and natural antibiotic for the treatment of oral pathogens: Abelmoschus esculentus green-formulated silver nanoparticles
  90. Optimization of gallic acid-enriched ultrasonic-assisted extraction from mango peels
  91. Effect of gamma rays irradiation in the structure, optical, and electrical properties of samarium doped bismuth titanate ceramics
  92. Combinatory in silico investigation for potential inhibitors from Curcuma sahuynhensis Škorničk. & N.S. Lý volatile phytoconstituents against influenza A hemagglutinin, SARS-CoV-2 main protease, and Omicron-variant spike protein
  93. Physical, mechanical, and gamma ray shielding properties of the Bi2O3–BaO–B2O3–ZnO–As2O3–MgO–Na2O glass system
  94. Twofold interpenetrated 3D Cd(ii) complex: Crystal structure and luminescent property
  95. Study on the microstructure and soil quality variation of composite soil with soft rock and sand
  96. Ancient spring waters still emerging and accessible in the Roman Forum area: Chemical–physical and microbiological characterization
  97. Extraction and characterization of type I collagen from scales of Mexican Biajaiba fish
  98. Finding small molecular compounds to decrease trimethylamine oxide levels in atherosclerosis by virtual screening
  99. Prefatory in silico studies and in vitro insecticidal effect of Nigella sativa (L.) essential oil and its active compound (carvacrol) against the Callosobruchus maculatus adults (Fab), a major pest of chickpea
  100. Polymerized methyl imidazole silver bromide (CH3C6H5AgBr)6: Synthesis, crystal structures, and catalytic activity
  101. Using calcined waste fish bones as a green solid catalyst for biodiesel production from date seed oil
  102. Influence of the addition of WO3 on TeO2–Na2O glass systems in view of the feature of mechanical, optical, and photon attenuation
  103. Naringin ameliorates 5-fluorouracil elicited neurotoxicity by curtailing oxidative stress and iNOS/NF-ĸB/caspase-3 pathway
  104. GC-MS profile of extracts of an endophytic fungus Alternaria and evaluation of its anticancer and antibacterial potentialities
  105. Green synthesis, chemical characterization, and antioxidant and anti-colorectal cancer effects of vanadium nanoparticles
  106. Determination of caffeine content in coffee drinks prepared in some coffee shops in the local market in Jeddah City, Saudi Arabia
  107. A new 3D supramolecular Cu(ii) framework: Crystal structure and photocatalytic characteristics
  108. Bordeaux mixture accelerates ripening, delays senescence, and promotes metabolite accumulation in jujube fruit
  109. Important application value of injectable hydrogels loaded with omeprazole Schiff base complex in the treatment of pancreatitis
  110. Color tunable benzothiadiazole-based small molecules for lightening applications
  111. Investigation of structural, dielectric, impedance, and mechanical properties of hydroxyapatite-modified barium titanate composites for biomedical applications
  112. Metal gel particles loaded with epidermal cell growth factor promote skin wound repair mechanism by regulating miRNA
  113. In vitro exploration of Hypsizygus ulmarius (Bull.) mushroom fruiting bodies: Potential antidiabetic and anti-inflammatory agent
  114. Alteration in the molecular structure of the adenine base exposed to gamma irradiation: An ESR study
  115. Comprehensive study of optical, thermal, and gamma-ray shielding properties of Bi2O3–ZnO–PbO–B2O3 glasses
  116. Lewis acids as co-catalysts in Pd-based catalyzed systems of the octene-1 hydroethoxycarbonylation reaction
  117. Synthesis, Hirshfeld surface analysis, thermal, and selective α-glucosidase inhibitory studies of Schiff base transition metal complexes
  118. Protective properties of AgNPs green-synthesized by Abelmoschus esculentus on retinal damage on the virtue of its anti-inflammatory and antioxidant effects in diabetic rat
  119. Effects of green decorated AgNPs on lignin-modified magnetic nanoparticles mediated by Cydonia on cecal ligation and puncture-induced sepsis
  120. Treatment of gastric cancer by green mediated silver nanoparticles using Pistacia atlantica bark aqueous extract
  121. Preparation of newly developed porcelain ceramics containing WO3 nanoparticles for radiation shielding applications
  122. Utilization of computational methods for the identification of new natural inhibitors of human neutrophil elastase in inflammation therapy
  123. Some anticancer agents as effective glutathione S-transferase (GST) inhibitors
  124. Clay-based bricks’ rich illite mineral for gamma-ray shielding applications: An experimental evaluation of the effect of pressure rates on gamma-ray attenuation parameters
  125. Stability kinetics of orevactaene pigments produced by Epicoccum nigrum in solid-state fermentation
  126. Treatment of denture stomatitis using iron nanoparticles green-synthesized by Silybum marianum extract
  127. Characterization and antioxidant potential of white mustard (Brassica hirta) leaf extract and stabilization of sunflower oil
  128. Characteristics of Langmuir monomolecular monolayers formed by the novel oil blends
  129. Strategies for optimizing the single GdSrFeO4 phase synthesis
  130. Oleic acid and linoleic acid nanosomes boost immunity and provoke cell death via the upregulation of beta-defensin-4 at genetic and epigenetic levels
  131. Unraveling the therapeutic potential of Bombax ceiba roots: A comprehensive study of chemical composition, heavy metal content, antibacterial activity, and in silico analysis
  132. Green synthesis of AgNPs using plant extract and investigation of its anti-human colorectal cancer application
  133. The adsorption of naproxen on adsorbents obtained from pepper stalk extract by green synthesis
  134. Treatment of gastric cancer by silver nanoparticles encapsulated by chitosan polymers mediated by Pistacia atlantica extract under ultrasound condition
  135. In vitro protective and anti-inflammatory effects of Capparis spinosa and its flavonoids profile
  136. Wear and corrosion behavior of TiC and WC coatings deposited on high-speed steels by electro-spark deposition
  137. Therapeutic effects of green-formulated gold nanoparticles by Origanum majorana on spinal cord injury in rats
  138. Melanin antibacterial activity of two new strains, SN1 and SN2, of Exophiala phaeomuriformis against five human pathogens
  139. Evaluation of the analgesic and anesthetic properties of silver nanoparticles supported over biodegradable acacia gum-modified magnetic nanoparticles
  140. Review Articles
  141. Role and mechanism of fruit waste polyphenols in diabetes management
  142. A comprehensive review of non-alkaloidal metabolites from the subfamily Amaryllidoideae (Amaryllidaceae)
  143. Discovery of the chemical constituents, structural characteristics, and pharmacological functions of Chinese caterpillar fungus
  144. Eco-friendly green approach of nickel oxide nanoparticles for biomedical applications
  145. Advances in the pharmaceutical research of curcumin for oral administration
  146. Rapid Communication
  147. Determination of the contents of bioactive compounds in St. John’s wort (Hypericum perforatum): Comparison of commercial and wild samples
  148. Retraction
  149. Retraction of “Two mixed-ligand coordination polymers based on 2,5-thiophenedicarboxylic acid and flexible N-donor ligands: The protective effect on periodontitis via reducing the release of IL-1β and TNF-α”
  150. Topical Issue on Phytochemicals, biological and toxicological analysis of aromatic medicinal plants
  151. Anti-plasmodial potential of selected medicinal plants and a compound Atropine isolated from Eucalyptus obliqua
  152. Anthocyanin extract from black rice attenuates chronic inflammation in DSS-induced colitis mouse model by modulating the gut microbiota
  153. Evaluation of antibiofilm and cytotoxicity effect of Rumex vesicarius methanol extract
  154. Chemical compositions of Litsea umbellata and inhibition activities
  155. Green synthesis, characterization of silver nanoparticles using Rhynchosia capitata leaf extract and their biological activities
  156. GC-MS analysis and antibacterial activities of some plants belonging to the genus Euphorbia on selected bacterial isolates
  157. The abrogative effect of propolis on acrylamide-induced toxicity in male albino rats: Histological study
  158. A phytoconstituent 6-aminoflavone ameliorates lipopolysaccharide-induced oxidative stress mediated synapse and memory dysfunction via p-Akt/NF-kB pathway in albino mice
  159. Anti-diabetic potentials of Sorbaria tomentosa Lindl. Rehder: Phytochemistry (GC-MS analysis), α-amylase, α-glucosidase inhibitory, in vivo hypoglycemic, and biochemical analysis
  160. Assessment of cytotoxic and apoptotic activities of the Cassia angustifolia aqueous extract against SW480 colon cancer
  161. Biochemical analysis, antioxidant, and antibacterial efficacy of the bee propolis extract (Hymenoptera: Apis mellifera) against Staphylococcus aureus-induced infection in BALB/c mice: In vitro and in vivo study
  162. Assessment of essential elements and heavy metals in Saudi Arabian rice samples underwent various processing methods
  163. Two new compounds from leaves of Capparis dongvanensis (Sy, B. H. Quang & D. V. Hai) and inhibition activities
  164. Hydroxyquinoline sulfanilamide ameliorates STZ-induced hyperglycemia-mediated amyleoid beta burden and memory impairment in adult mice
  165. An automated reading of semi-quantitative hemagglutination results in microplates: Micro-assay for plant lectins
  166. Inductively coupled plasma mass spectrometry assessment of essential and toxic trace elements in traditional spices consumed by the population of the Middle Eastern region in their recipes
  167. Phytochemical analysis and anticancer activity of the Pithecellobium dulce seed extract in colorectal cancer cells
  168. Impact of climatic disturbances on the chemical compositions and metabolites of Salvia officinalis
  169. Physicochemical characterization, antioxidant and antifungal activities of essential oils of Urginea maritima and Allium sativum
  170. Phytochemical analysis and antifungal efficiency of Origanum majorana extracts against some phytopathogenic fungi causing tomato damping-off diseases
  171. Special Issue on 4th IC3PE
  172. Graphene quantum dots: A comprehensive overview
  173. Studies on the intercalation of calcium–aluminium layered double hydroxide-MCPA and its controlled release mechanism as a potential green herbicide
  174. Synergetic effect of adsorption and photocatalysis by zinc ferrite-anchored graphitic carbon nitride nanosheet for the removal of ciprofloxacin under visible light irradiation
  175. Exploring anticancer activity of the Indonesian guava leaf (Psidium guajava L.) fraction on various human cancer cell lines in an in vitro cell-based approach
  176. The comparison of gold extraction methods from the rock using thiourea and thiosulfate
  177. Special Issue on Marine environmental sciences and significance of the multidisciplinary approaches
  178. Sorption of alkylphenols and estrogens on microplastics in marine conditions
  179. Cytotoxic ketosteroids from the Red Sea soft coral Dendronephthya sp.
  180. Antibacterial and biofilm prevention metabolites from Acanthophora spicifera
  181. Characteristics, source, and health risk assessment of aerosol polyaromatic hydrocarbons in the rural and urban regions of western Saudi Arabia
  182. Special Issue on Advanced Nanomaterials for Energy, Environmental and Biological Applications - Part II
  183. Green synthesis, characterization, and evaluation of antibacterial activities of cobalt nanoparticles produced by marine fungal species Periconia prolifica
  184. Combustion-mediated sol–gel preparation of cobalt-doped ZnO nanohybrids for the degradation of acid red and antibacterial performance
  185. Perinatal supplementation with selenium nanoparticles modified with ascorbic acid improves hepatotoxicity in rat gestational diabetes
  186. Evaluation and chemical characterization of bioactive secondary metabolites from endophytic fungi associated with the ethnomedicinal plant Bergenia ciliata
  187. Enhancing photovoltaic efficiency with SQI-Br and SQI-I sensitizers: A comparative analysis
  188. Nanostructured p-PbS/p-CuO sulfide/oxide bilayer heterojunction as a promising photoelectrode for hydrogen gas generation
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Heruntergeladen am 29.9.2025 von https://www.degruyterbrill.com/document/doi/10.1515/chem-2022-0358/html
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