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Purification and characterization of glucose-6-phosphate dehydrogenase from Eisenia fetida and effects of some pesticides and metal ions

  • Naciye Kayhan , Veysel Çomaklı , Sevki Adem ORCID logo EMAIL logo and Caglar Güler
Published/Copyright: May 18, 2020
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Turkish Journal of Biochemistry
From the journal Turkish Journal of Biochemistry Volume 45 Issue 4

Abstract

Objectives

Earthworms have a large impact on the soil ecosystem. They are quite sensitive to pollutants. Purification and biochemical characterization of glucose-6-phosphate dehydrogenases (G6PD) from the earthworm species Eisenia fetida were aimed. The determination of the toxicity potentials of some soil pollutants on G6PD activity was intended.

Methods

G6PD was isolated using 2′,5′-ADP-Sepharose 4B affinity column. Enzyme purity and molecular mass were determined by SDS-PAGE. Its biochemical properties investigated. The effects of some soil pollutants on the enzyme were studied in vitro.

Results

Enzyme was purified with 28% yields and 232 fold. Optimum pH and buffer concentration, optimal and stable temperature was determined as pH: 8.5, 60 mM, 25 °C and 20 °C. Its molecular weight estimated as 36 kDa. The Ni2+, Hg2+, Pb2+, Cr2+, and Fe2+ ions with IC50 values in the range of 56 ± 06−120 ± 20 μM and the diniconazole, metalaxyl, methomyl, carboxyl, and oxamyl with IC50 values in the range of 7.6 ± 1.2−77 ± 12 μM exhibited an inhibitory effect on G6PD.

Conclusions

G6PD was isolated and characterized from E. fetida. Its catalytic activity decreased with very low concentration by pesticides and metal ions. The results indicated that the inhibition of G6PD may be important in the toxicity mechanism of pollutants on this earthworm.

Öz

Amaç

Toprak solucanlarının toprak ekosistemi üzerinde büyük etkisi vardır. Kirleticilere karşı oldukça hassastırlar. Toprak solucanı türlerinden Eisenia fetida'dan glikoz-6-fosfat dehidrogenazın (G6PD) saflaştırılması ve biyokimyasal karakterizasyonu hedeflenmiştir. Bazı toprak kirleticilerinin, G6PD aktivitesi üzerindeki toksisite potansiyellerinin belirlenmesi amaçlanmıştır.

Gereç ve Yöntem

G6PD, 2′,5′-ADP-Sepharose 4B afinite kolonu kullanılarak izole edildi. Enzim saflığı ve molekül kütle SDS-PAGE ile belirlendi. Biyokimyasal özellikleri araştırıldı. Bazı pestisitlerin ve metal iyonlarının enzim üzerindeki etkileri in vitro incelenmiştir.

Bulgular

Enzim % 28 verimle 232 kat saflaştırıldı. Optimum pH ve tampon konsantrasyonu, optimal ve stabil sıcaklık pH: 8.5, 60 mM, 25 °C ve 20 °C olarak belirlenmiştir. Molekül kütlesi 36 kDa olarak tespit edildi. Ni2+, Hg2+, Pb2+, Cr2+, ve Fe2+ 56 ± 06−120 ± 20 μM aralığında IC50 değerleri ile diniconazole, metalaxyl, methomyl, carboxyl, ve oxamyl 7.6 ± 1.2−77 ± 12 μM aralığında IC50 değerleri ile G6PD üzerinde inhibisyon etkisi gösterdi.

Sonuç

G6PD, E. fetida'dan saflaştırıldı ve karakterize edildi. Onun katalitik etkinliği, pestisitler ve metal iyonları tarafından çok düşük konsantrasyonlarda azaltılmıştır. Sonuçlar, bu solucan üzerinde, kirleticilerin toksisite mekanizmasında G6PD'nin inhibisyonunun önemli olabileceğini göstermektedir.

Keywords: Eisenia fetida; glucose-6-phosphate dehydrogenase; heavy metal; pesticide; toxic
Anahtar Kelimeler: Eisenia fetida; glikoz-6-fosfat dehidrojenaz; ağır metal; pestisit; toksik

Introduction

In recent years, soil pollution resulting from agricultural activities and industrial development has enhanced the interest in studies related to this field [1], [2]. As important pollutants, petroleum and petroleum products, chlorophenols, aromatic hydrocarbons, pesticide, and heavy metal ions entering the soils and waters directly or indirectly pose a violent threat to human health and the natural ecosystem [3], [4], [5], [6]. Earthworms that live in the soils play an essential role in the performance of the lands exposed to the effects of pesticides and heavy metals, which are prevalent on the grounds due to agricultural and industrial activities [7], [8]. The biochemical responses of worms to environmental stress are sensitive and informative. The chemical toxicity depends on the concentration and application time of the toxic substances. The toxic substances in the living organism cause tissue damage and reducing resistance to diseases. As a result, their effects result in death. Hence, the biochemical responses of the worms to toxic substances in soils can serve as an early indicator of soil pollution [9].

Glucose 6-phosphate dehydrogenase (EC 1.1.1.49-d-glucose-6-phosphate: NADP+ 1-oxidoreductase) is the first and critical enzyme in the pentose phosphate metabolic pathway. It is responsible for the synthesis of ribose-5- phosphate, and NADPH. The first plays a role in the synthesis of nucleic acids, while the second participates in reductive biosynthesis reactions and detoxification mechanisms [10], [11]. Previous studies showed that the worms exposed to the toxic effects of environmental pollutants produce reactive oxygen species (ROS) which damage the macromolecules such as nucleic acids, proteins, and lipids, which in turn causes cell damage [12], [13], [14]. One of the most critical factors in increasing the amount of reactive oxygen is the inhibition of antioxidant enzymes by pollutants. Glucose-6-phosphate dehydrogenase (G6PD) is an essential enzyme that provides the high amount of NADPH used as reducing agent in the glutathione and cytochrome P450 the essential antioxidant systems in the cell, and it protects the cells against chemical and oxidative damages [15]. It is therefore important to determine the effects of environmental pollutants on this enzyme. Many investigators have reported the effects of metal ions and pesticides on enzyme activities [10], [16], [17]. Also, many studies were reported toxic effects on the antioxidant enzyme activities of the heavy metals and pesticide toward earthworm species [18]. However, there is no study in the literature indicating the effect of these soil pollutants in the soil worm G6PD.

In the present study was aimed that the G6PD enzyme was purified from E. fetida, an earthworm species referred to as “the ecosystem engineer” by the local community because of its role in the efficiency of the soil structure. Besides, following the characterization of the purified enzyme, the toxic effects of some pesticides and heavy metal ions were investigated.

Materials and methods

Materials

The protein standards and chemicals were obtained from Sigma Chem. (Germany, Hamburg), and E. Merck (Germany, Taufkirchen). Affinity material was purchased from GE Healthcare (USA, Marlborough). The worms were taken from the Biology Department of Science Faculty of the Cankiri Karatekin University.

Homogenate preparation and enzyme purification

Worms were kept in pure water for 12 h to remove the soil within. They were stored at −20 °C until the experimental studies. To prepare homogenate, they were thoroughly crushed in the porcelain mortar, and frozen and resolved several times. The sample was transferred into chilled buffer including 1 mM EDTA, 2 mM dithiothreitol (DTT) and 20 mM Tris-HCl (pH = 7.5). Homogenate was centrifuged at 11,000 × g for 15 min at + 4 °C. The supernatant was carefully removed. Then it filtered using filter paper to remove particles that may be in the supernatant

The supernatant was loaded onto 2′5′ ADP Sepharose™ 4B media column arranged previously the defined protocol [19]. Then washing buffer containing 50 mM phosphate, 1 mM DTT, 1 mM EDTA, pH 7.35 was passed from affinity column until the absorbance difference between the applied and eluted samples is 0.025 at 280 nm. This process took up approximately 4 h. Before elution, a 10 mL elution buffer without NADP+ was passed through the column. The column-bound G6PD was eluted by using 20 mL buffer including 80 mM phosphate, 80 mM KCl, 1 mM NADP+, 1 mM EDTA, pH 7.85 at 1 mL aliquots. Active tubes were combined and dialyzed against buffer 20 mM cold phosphate buffer, pH 7.5 at + 4 °C for 24 h.

To prepare the purification table, the enzyme activity, total volume, and protein amount were determined at each step. The amount of protein in all steps was calculated using to the Bradford method using as a standard the BSA [20]. SDS-PAGE described by Laemmli was made to check the purity of the enzyme and determine the molecular masses of the subunits, using markers containing between 6.5 and 212 kDa as standard [21].

Activity assay

Enzyme activity assay was performed according to Beutler's method based on measuring the change in absorbance at 340 nm in a 1,000 μL reaction volume at 25 °C on the Rayleigh-UV-2601 UV/VIS Spectrophotometer [22]. The standard enzyme assay included 60 μM Tris-HCl, pH 8.5, 0.6 mM G6P and 0.2 mM NADP+. Each G6PD activity values detected with triplicate trials.

Biochemical characterization

The dependence of G6PD on pH was detected in 0.1 M phosphate buffer, pH 6–7.5, 0.1 M Tris-HCl, pH 7.5–9.5. The optimal buffer concentration for the enzyme activity was determined to be in the range of 10–90 mM Tris-HCl.

The effect of temperature on the catalytic activity of the enzyme was determined by measuring the enzyme activity at 15, 25, 30, 40, 50, and 60 °C. Also, the stable temperature for the enzyme was evaluated waiting for 30 min at 20, 30, 40, and 50 °C.

The kinetic parameters of purified enzyme were figured out under enzyme assay conditions, in the presence of different NADP+ (4–64 μM) and G6P (10–200 μM) concentrations. KM and Vmax values for NADP+ and G6P were determined using Lineweaver–Burk graphs. We also investigated if NADPH the end product of the reaction catalyzed by the purified enzyme inhibits the binding of NADP+ and G6P to glucose-6-phosphate dehydrogenase. Five varied NADP+ (4, 8, 12, 16, and 20 μM) and G6P (12, 24, 36, 48, and 60 μM) concentrations with enzyme solution were incubated with three NADPH concentrations (10, 15, and 25 μM). Lineweaver-Burk plots of 1/V versus 1/NADP+ and 1/G6P were used to analyze the G6PD inhibition pattern of NADPH.

Inhibition studies of metal ions and pesticides

To determine the effects of some ion of metal and pesticide on the G6PDenzyme activity, enzyme activity was tested at least five different concentrations as in vitro. Stock solutions of metal ions were prepared from their salts (Pb(NO3)2, Cr(NO3)2, HgCl2, FeSO4, Ni(NO3)2) as 5 mg/mL, then diluted using pure water. In inhibition studies, metal ions were used in the concentration range of 1–250 μM. The oxamyl (1–200 μM), carboryl (1–40 μM), diniconazole (1–20 μM), metalaxyl (1–18 μM), methomyl (1–20 μM), carbofuran (1–200 μM), tebuconazole (1–250 μM), atrazine (1–200 μM), propoxur (1–200 μM),1- naphthol (1–200 μM), and 2,4–D(1–200 μM) pesticides, simazine herbicide were first dissolved in DMSO as 1 mg/mL, then diluted 10-fold with distilled water. Inhibitor-free enzyme activity was accepted as a control. The inhibitor concentrations decreasing enzyme activity by 50% were calculated from percent activity-concentration graphs drawing in Excel 2010. Ki values of metal ions and pesticides exhibited inhibitory effect was calculated by the Cheng–Prusoff equation [23].

Results

In the present study, after the purification of the G6PD enzyme from the earthworm species E. fetida and its characterization, the toxic effects of some pesticides and heavy metal ions on the activity of the enzyme were investigated. Using the 2′,5′-ADP sepharose-4B affinity chromatography, the protein was successfully purified in a single step. Table 1 shows that the purification method achieved a 232-fold purification of the G6PD enzyme from E. fetida with a specific activity of 11.63 E U/mg proteins and a recovery of 28%. The purity of the enzyme was tested with the SDS-PAGE method (Figure 1). The molecular mass was determined to be 36 kDa by SDS-PAGE under denaturing conditions.

Table 1:

Purification table of G6PD from E. fetida.



Purification steps		Total volume, mLActivity, U/mLTotal activity, UAmount of protein, mg/mLSpecific activity, U/mg% yieldPurification fold
The homogenates500.1335.62.2400.0501001
Affinity chromatography90.1771.50.01511.63528232
Figure 1: SDS-PAGE bands of E. fetida G6PD enzyme.
Figure 1:

SDS-PAGE bands of E. fetida G6PD enzyme.

The biochemical properties of the purified enzyme such as optimum pH, stable pH, buffer concentration activity, optimum temperature, and thermostability were characterized (Figure 2A–D). Then, using the Lineweaver-Burk plot, the affinity of the enzyme to its substrates was analyzed at the optimum conditions determined in the study (optimum pH, optimum buffer concentration activity, and optimum temperature). For the G6P and NADP+ substrates, the KM values were 0.14 and 0.20 mM, respectively, while the Vmax values were 0.374 and 0.525 EU/mL, respectively.

Figure 2: Optimal conditions for G6PD from E. fetida.(A). The effect of pH on enzyme activity, (B). The effect of buffer concentration on enzyme activity, (C). The effect of temperature on enzyme activity, (D). Thermal stability of enzyme.
Figure 2:

Optimal conditions for G6PD from E. fetida.

(A). The effect of pH on enzyme activity, (B). The effect of buffer concentration on enzyme activity, (C). The effect of temperature on enzyme activity, (D). Thermal stability of enzyme.

NADPH inhibited the G6PD enzyme with an IC5O value of approximately 150 ± 10 μM. The inhibition type of NADPH determined with Lineweaver–Burk graphs (Figure 3A, B). While NADPH blocked as uncompetitive the binding of substrate G6P to the G6PD enzyme with Ki values 0.211 ± 0.06 μM, it inhibited as uncompetitive the binding of coenzyme NADPH to the G6PD enzyme with Ki values 0.179 ± 0.08 μM.

Figure 3: Effect of NADPH on the binding of NADP+ and G6P to G6PG.(A). Lineweaver–Burk graph mechanism of competitive type for NADPH with respect to NADP+, (B). Lineweaver–Burk graph mechanism of uncompetitive type for NADPH with respect to G6P.
Figure 3:

Effect of NADPH on the binding of NADP+ and G6P to G6PG.

(A). Lineweaver–Burk graph mechanism of competitive type for NADPH with respect to NADP+, (B). Lineweaver–Burk graph mechanism of uncompetitive type for NADPH with respect to G6P.

The inhibitory effect of some metal ions and pesticides on the G6PD enzyme activity of the E. fetida earthworm was also investigated. All of the selected metal ions and the oxamyl, carboxyl, diniconazole, metalaxyl, and methomyl pesticides show inhibitory effects on enzyme activity. IC50 values were calculated from % activity – [inhibitory concentration] graphs (Figure 4A–D). As seen in Table 2, the IC50 values of the Pb+2, Cr+2, Hg+2, Fe+2 and Ni+2 metal ions were 109 ± 14, 69 ± 6, 110 ± 15, 120 ± 20, and 56 ± 06 μM, respectively. As seen in Table 3, the IC50 values of the oxamyl, carboxyl, diniconazole, metalaxyl, and methomyl pesticides were 77 ± 12, 13 ± 2.6, 7.6 ± 1.2, 10 ± 0.9, and 11 ± 0.8 μM, respectively. Ki values obtained from the Cheng–Prusoff equation were represented in Table 3.

Figure 4: Inhibition graphs of some pesticides and metal ions on Eisenia fetida G6PD enzyme.(A). The concentration-dependent inhibition effect of Ni+2 on enzyme activity, (B). The concentration-dependent inhibition effect of Hg+2 on enzyme activity, (C). The concentration-dependent inhibition effect of Diniconazole on enzyme activity, (D). The concentration-dependent inhibition effect of Metalaxyl on enzyme activity.
Figure 4:

Inhibition graphs of some pesticides and metal ions on Eisenia fetida G6PD enzyme.

(A). The concentration-dependent inhibition effect of Ni+2 on enzyme activity, (B). The concentration-dependent inhibition effect of Hg+2 on enzyme activity, (C). The concentration-dependent inhibition effect of Diniconazole on enzyme activity, (D). The concentration-dependent inhibition effect of Metalaxyl on enzyme activity.

Table 2:

IC50 and Ki values for metal ions inhibiting to G6PD from E. fetida.

Metal IonsIC50 values, μMKi constant concerning G6P, μMKi constant concerning NADP+, μM
Ni2+56 ± 610.59 ± 1.328.00 ± 3.2
Hg2+69 ± 813.59 ± 1.534.50 ± 4.0
Pb2+109 ± 1420.62 ± 2.654.50 ± 7.1
Cr2+110 ± 1520.81 ± 2.855.00 ± 7.5
Fe2+120 ± 2022.70 ± 3.760.00 ± 10.0
Table 3:

IC50 and Ki Values for pesticides inhibiting to G6PD from E. fetida.

PesticidesIC50 values, μMKi constant concerning G6P, μMKi constant concerning NADP+, μM
Diniconazole7.6 ± 1.21.44 ± 0.23.80 ± 0.6
Carboryl13.0 ± 2.62.46 ± 0.56.50 ± 1.3
Metalaxyl10.0 ± 0.91.89 ± 0.25.00 ± 0.4
Methomyl11.0 ± 0.82.08 ± 0.15.50 ± 0.4
Oxamyl77.0 ± 12.014.57 ± 2.238.50 ± 6.0

Discussion

The purification of the G6PD enzyme from the E. fetida was conducted using the 2′,5′-ADP sepharose-4B affinity chromatography. In previous studies, the purification process was performed using multi-chromatographic methods that involved different combinations of affinity, ion exchange, and gel filtration chromatography [19]. 2′,5′-ADP sepharose-4B is the most preferred affinity column material for the purification of NADP+/H-dependent enzymes. This material offers advantages such as rapid, reusable, less chemical use, and fewer steps of purification. We have successfully purified the enzyme in the single-step using this material. This and previous studies indicate that this affinity material is suitable material to purify NADP+/H-dependent enzymes [10], [11], [15]. The purification coefficient and purification efficiency of the enzyme are very low compared to those reported from other studies [11], [15]. This may be because the enzyme has low affinity against to NADP+. The molecular mass of the purified enzyme was determined to be 36 kDa using SDS-PAGE. Similar results have been reported in previous articles [24], [25].

The enzyme activities are affected by changes from factors such as pH, buffer concentration, and temperature. In literature, the pH value at which the enzyme has the highest activity was reported to be within the range of 7.0–9.5 [10], [11], [26], [27]. The optimum pH value for the E. fetida G6PD enzyme was determined to be 8.5, which agrees with the results reported in the relevant literature. The optimum buffer concentration activity of the enzyme at pH 8.5 in tris-HCl buffer was 60 mM. A review of previous studies showed that the G6PD enzyme of animal origin nearly loses its activity at temperatures around 55–60 °C [15], [27], [28]. The optimum temperature of the purified enzyme was 25 °C, and its thermostability temperature was 20 °C. When Figure 2 is examined, it is observed that the enzyme loses significant activity as temperature increases. The optimal temperature determined for the enzyme is in parallel with the optimum growth rate for the E. fetida species [29].

KM values are an indicator of enzyme-substrate interaction. These values may vary depending on the source of the enzyme and the optimum conditions. For the G6P and NADP+ substrates, the KM values were 0.14 and 0.20 mM, respectively. According to the values, the KM value for G6P was lower than the value for NADP+. This indicates that the enzyme had a higher affinity for G6P. The values contradict to the results reported in previous studies for the G6PD obtained from different sources [30], [31], [32].

Control point enzymes in metabolism were generally inhibited by their product. G6PD is the first control point enzyme in the oxidative branch of the hexose monophosphate shunt. It has been known that G6PD activity is balanced with the level of NADPH in the cell, so NADPH is the most important regulator of the pentose pathway [15], [33], [34]. Ki constants calculated for E. fetida G6PD are lower than rat lung and rainbow trout (Oncorhynchus mykiss) erythrocytes [15], [28], but higher than Taenia crassiceps and Escherichia coli [35], [36].

Furthermore, the inhibitory effect of some metal ions and pesticides on the G6PD enzyme activity of the E. fetida earthworm was also investigated. Heavy metals and pesticides are known to have different toxicological effects on organisms. Although previous studies have investigated the toxic effects of both metal ions and pesticides on the important metabolic enzymes in living organisms, their in vitro effects on the G6PD enzyme activity of the E. fetida earthworm are yet to be investigated. All of the selected metal ions and the oxamyl, carboxyl, diniconazole, metalaxyl, and methomyl pesticides show inhibitory effects on enzyme activity. Ki constants and IC50 values are the best parameters to determine inhibitory effects. IC50 values of metal ions were the range of 56 ± 06−110 ± 15 μM. As seen in the IC50 values of the pesticides were the ranges of 7.6 ± 1.2−77 ± 12 μM. Pesticides had this effect at even lower concentrations. Ki constants were determined for each substrate. Both the compatibility of the Ki constants with the IC50 values and obtaining a lower Ki constant in the G6P substrate with a lower KM value is highly significant. Similar results were obtained in previous studies conducted under both in vitro and in vivo conditions[10], [37]

According to the data given in Tables 2 and 3, all of the investigated substances were powerful inhibitors of the G6PD enzyme of the E. fetida earthworm. Specifically, the Ni+2 metal ion with a Ki value of 10.59 μM and the diniconazole pesticide with a Ki value of 1.44 μM had the highest inhibitory effects.

The results obtained in the study can contribute to the reviews on the biology, biochemistry, and toxicology of natural organisms and sustainable agriculture. Pesticides and heavy metals are prevalent pollutants in the soils that stem from various industrial and agricultural activities [38], [39]. The response of the metabolism to these pollutants should be well defined. Therefore, the changes in the G6PD enzyme activity due to pesticides and metal ions and the resulting undesired side effects on NADPH synthesis can be evaluated by referring to the Ki and IC50 values obtained in the study.

Conclusion

Earthworms are evaluated keystone organisms in adjusting nutrient cycling processes in soil. Similarly, G6PD has the central role in antioxidant defense, nucleotide precursor synthesis, and lipid synthesis. G6PD from ecotoxicologically important earthworm species E. fetida successfully isolated for the first time using 2′5′ ADP Sepharose™ 4B. Its biochemical properties characterized. The effect of some important soil pollutants on enzyme activity was investigated. According to Ki values, metal ions have a high inhibitory effect on the enzyme. In particular, they inhibit the binding of G6P to the enzyme with Ki values under 22 μM. Pesticides such as diniconazole, carboryl, metalaxyl, and methomyl inhibited the binding of both NADP+ and G6P to the enzyme Ki values under 7 μM. The results show that the inhibition of the G6PD enzyme may be important in the mechanism of the toxicity effect of metal ions and pesticides on earthworm species E. fetida. Future studies related to pesticide and metal ion toxicity on worms may add this enzyme activity measurement to the parameters of toxicity.

Corresponding author: Dr. Sevki Adem, Department of Chemistry, Cankiri Karatekin University, Faculty of Science, 18100, Cankiri, Turkey, E-mail:

Funding source: Çankiri Karatekin Üniversitesi

Award Identifier / Grant number: FF200217B42

Acknowledgments

This research was supported by Cankiri Karatekin University (Project No: FF200217B42).

  1. Competing interests: The authors declare no conflict interest.

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Received: 2019-04-11
Accepted: 2019-12-06
Published Online: 2020-05-18

© 2020 Walter de Gruyter GmbH, Berlin/Boston

Articles in the same Issue

  1. Frontmatter
  2. Research Articles
  3. Development of new total RNA isolation method for tissues with rich phenolic compounds
  4. Myofibrillar degeneration with diphtheria toxin
  5. In vitro and in silico studies on AChE inhibitory effects of a series of donepezil-like arylidene indanones
  6. In vitro antioxidant, anti-inflammatory and anti-cancer activities of methanolic extract of Asparagus horridus grows in North Cyprus Kuzey Kıbrıs da yetişen Asparagus horridus metanolik ekstraktının in-vitro antioksidan, anti-enflamatuar ve anti-kanser aktivitesi
  7. Purification and characterization of glucose-6-phosphate dehydrogenase from Eisenia fetida and effects of some pesticides and metal ions
  8. Nephroprotective effects of eriocitrin via alleviation of oxidative stress and DNA damage against cisplatin-induced renal toxicity
  9. The impact of orally administered gadolinium orthovanadate GdVO4:Eu3+ nanoparticles on the state of phospholipid bilayer of erythrocytes
  10. An anxiolytic drug buspirone ameliorates hyperglycemia and endothelial dysfunction in type 2 diabetic rat model
  11. Effects of mesenchymal stem cell and amnion membrane transfer on prevention of pericardial adhesions
  12. How potential endocrine disruptor deltamethrin effects antioxidant enzyme levels and total antioxidant status on model organisms
  13. Antiproliferative effect of rosehip tea phenolics in prostate cancer cell lines
  14. Investigation of MMP-9 rs3918242 and TIMP-2 rs8179090 polymorphisms in renal cell carcinoma tissues
  15. Investigation of SR-BI gene rs4238001 and rs5888 polymorphisms prevalence and effects on Turkish patients with metabolic syndrome
  16. Assessment of the frequency and biochemical parameters of conjunctivitis in COVID-19 and other viral and bacterial conditions
  17. Short Communication
  18. Lack of hotspot mutations other than TP53 R249S in aflatoxin B1 associated hepatocellular carcinoma
  19. Letter to the Editors
  20. Cornuside, identified in Corni fructus, suppresses melanin biosynthesis in B16/F10 melanoma cells through tyrosinase inhibition
  21. The extract of male bee and beehive from Bombus terrestris has biological efficacies for promoting skin health
  22. COVID-19 laboratory biosafety guide
  23. Retraction note
https://doi.org/10.1515/tjb-2019-0156
Keywords for this article
Eisenia fetida; glucose-6-phosphate dehydrogenase; heavy metal; pesticide; toxic

Articles in the same Issue

  1. Frontmatter
  2. Research Articles
  3. Development of new total RNA isolation method for tissues with rich phenolic compounds
  4. Myofibrillar degeneration with diphtheria toxin
  5. In vitro and in silico studies on AChE inhibitory effects of a series of donepezil-like arylidene indanones
  6. In vitro antioxidant, anti-inflammatory and anti-cancer activities of methanolic extract of Asparagus horridus grows in North Cyprus Kuzey Kıbrıs da yetişen Asparagus horridus metanolik ekstraktının in-vitro antioksidan, anti-enflamatuar ve anti-kanser aktivitesi
  7. Purification and characterization of glucose-6-phosphate dehydrogenase from Eisenia fetida and effects of some pesticides and metal ions
  8. Nephroprotective effects of eriocitrin via alleviation of oxidative stress and DNA damage against cisplatin-induced renal toxicity
  9. The impact of orally administered gadolinium orthovanadate GdVO4:Eu3+ nanoparticles on the state of phospholipid bilayer of erythrocytes
  10. An anxiolytic drug buspirone ameliorates hyperglycemia and endothelial dysfunction in type 2 diabetic rat model
  11. Effects of mesenchymal stem cell and amnion membrane transfer on prevention of pericardial adhesions
  12. How potential endocrine disruptor deltamethrin effects antioxidant enzyme levels and total antioxidant status on model organisms
  13. Antiproliferative effect of rosehip tea phenolics in prostate cancer cell lines
  14. Investigation of MMP-9 rs3918242 and TIMP-2 rs8179090 polymorphisms in renal cell carcinoma tissues
  15. Investigation of SR-BI gene rs4238001 and rs5888 polymorphisms prevalence and effects on Turkish patients with metabolic syndrome
  16. Assessment of the frequency and biochemical parameters of conjunctivitis in COVID-19 and other viral and bacterial conditions
  17. Short Communication
  18. Lack of hotspot mutations other than TP53 R249S in aflatoxin B1 associated hepatocellular carcinoma
  19. Letter to the Editors
  20. Cornuside, identified in Corni fructus, suppresses melanin biosynthesis in B16/F10 melanoma cells through tyrosinase inhibition
  21. The extract of male bee and beehive from Bombus terrestris has biological efficacies for promoting skin health
  22. COVID-19 laboratory biosafety guide
  23. Retraction note
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