The Effect of Altitude on Phenolic, Antioxidant and Fatty Acid Compositions of Some Turkish Hazelnut (Coryllus avellana L.) Cultivars.

Turkey is the leading producer and exporter of hazelnuts, producing approximately 64% of global hazelnut production. This research investigated the effects of cultivars and altitude on the phenolic, antioxidant, and fatty acid compositions of five hazelnut cultivars grown at three different altitudes, 100 m, 350 m, and 800 m, in Ordu province, one of the territories that produce the most hazelnuts. The results showed that the cultivar and location significantly affected phenolic compounds, antioxidant activity, and fatty acid (FA) content. The lowest (2.30 mg/kg-Yagli) and highest (21.11 mg/kg-Kara) gallic acids were obtained at 100 m. The highest total phenolic content and antioxidant activity were found in the nuts grown at 350 m in the Kara and Palaz cultivars, at 100 m in the Yagli and Sivri cultivars, and at 800 m in the Çakildak cultivar. Oleic acid was the predominant FA in the cultivars and possessed a diverse trend according to the altitude and cultivar, ranging from 76.04% to 84.80%, increasing with altitude in all cultivars except Çakildak. Palmitic acid was the predominant saturated FA followed by stearic acid, which significantly varied according to the elevations. This study suggests that the responses of hazelnuts to altitude depend on the cultivar; hence, a proper approach to producing nuts containing more phenolic, fatty acids, and antioxidant activity includes choosing a suitable cultivar for a specific elevation.

A novel method for explaining the product inhibition mechanisms via molecular docking: inhibition studies for tyrosinase from Agaricus bisporus.

The present study aims to investigate the substrate (4-methyl catechol and catechol) specificity and inhibition mechanisms (l-ascorbic acid, citric acid, and l-cysteine) of the tyrosinase enzyme (TYR), which is held responsible for browning in foods and hyperpigmentation in the human skin, through kinetic and molecular docking studies. During the experimental studies, the diphenolase activities of TYR were determined, following which the inhibitory effects of the inhibitors upon the diphenolase activities of TYR. The inhibition types were determined as competitively for l-ascorbic acid and citric acid and noncompetitive for l-cysteine. The kinetic results showed that the substrate specificity was better for catechol while l-cysteine showed the best inhibition profile. As for the in silico studies, they also showed that catechol had a better affinity in line with the experimental results of this study, considering the interactions of the substrates with TYR's active site residues and their distance to CuB metal ion, which is an indicator of diphenolase activity. Besides, the inhibitory mechanisms of the inhibitor molecules were explained by the molecular modeling studies, considering the binding number of the inhibitors with the active site amino acid residues of TYR, the number and length of H bonds, negative binding energy values, and their distance to CuB metal ion. Based on our results, we suggest that the novel method used in this study to explain the inhibitory mechanism of l-cysteine may provide an affordable alternative to the expensive methods available for explaining the inhibitory mechanism of TYR and those of other enzymes. HighlightsThe best affinity for the tyrosinase enzyme occurred with catechol.l-Ascorbic acid, citric acid, l-cysteine inhibited the diphenolic activity of tyrosinase.In silico studies confirmed the best affinity shown by catechol.Product inhibition mechanism of l-cysteine explained by in silico for the first time.Communicated by Ramaswamy H. Sarma.

Purification and biochemical characterization of polyphenol oxidase extracted from Kirmizi Kismis grape (Vitis vinifera L.).

The purification of the polyphenol oxidase (PPO) enzyme from Kirmizi Kismis grape (Vitis vinifera L.) was performed 61.23 times using affinity chromatography. The molecular weight of the enzyme was found to be about 38.1 kDa by SDS-PAGE as a single band. The optimum pH and temperature values were revealed to be 5.0 and 30°C, respectively, in the presence of 4-methyl catechol substrate. The thermal stability of PPO was examined and it was observed to maintain its activity at 20°C for 1 hr. Km and Vmax values were determined to be 4.8 mM and 2000,0 EU/ml for 4-methyl catechol as a substrate. IC50 and Ki values and inhibition types were found for various browning agents and ascorbic acid had the strongest inhibitory impact on PPO. The inhibitory impact of Na+ , K+ , Mg2+ , Cu2+ , and Al3+ metal ions on the enzyme activity at final concentrations of 1 mM and 10 mM was examined. PRACTICAL APPLICATIONS: Grapes grown and processed take a significant place in our life. The grape has antioxidant, anticarcinogenic, antidiabetic and protective properties against bacteria and viruses. Furthermore, it takes an important position in the country's economy and social life due to providing raw materials to the food industry and having high export potential. Polyphenol oxidase, which is the leading actor of enzymatic browning reactions causing serious economic losses every year, was purified and characterized from Kirmizi Kismis grape (Vitis vinifera L.). This ancient grape variety has industrial processing and export potential due to its long storage life and resistance to oxidation. Therefore, the purification and biochemical characterization of polyphenol oxidase from Kirmizi Kismis grape are of great importance.

An Innovator Support Material for Tyrosinase Immobilization: Antimony-Doped Tin Oxide Thin Films (ATO-TF).

Tyrosinase (T3824, Sigma) enzymes were immobilized onto SnO2: Sb thin films (ATO-TF) which were synthesized in laboratory conditions by spray pyrolysis technique. Immobilization of tyrosinase onto SnO2: Sb thin film (ATO-TF) was confirmed by scanning electron microscopy (SEM) and Fourier transformed infrared spectroscopy (FTIR). The optimum pH of the immobilized tyrosinase (tyrosinase-ATO-TF) was 6.0, and the optimum temperature was found to be 30 °C. In the presence of catechol substrate of immobilized enzymes, Km and Vmax values were determined as 0.34 mM and 312.5 U/cm2 min, respectively. The thermal stabilities of the immobilized tyrosinase at 4 °C and 30 °C were investigated for 20, 40, and 60 min. At these temperatures and time intervals, the immobilized tyrosinase was found to be highly stable. The pH stability of the immobilized tyrosinase enzymes was incubated for 24, 48, 72, and 96 h at 4 °C temperature. The enzyme activity was determined under optimum conditions, and the activity of the immobilized tyrosinase enzymes exhibited various values for the range of pH 3.0-8.0. The storage stability of the immobilized tyrosinase enzymes at 4 °C was investigated, and 45.72% of the initial activity was maintained at the end of the seventh day. Furthermore, the reusability of the immobilized tyrosinase enzymes has been examined. The immobilized tyrosinase enzymes can be used 3 times and survived their 50% of initial activity. The ATO-TF can be used as alternative support materials for the immobilization of tyrosinase enzymes.

The toxicological impacts of some heavy metals on carbonic anhydrase from gilthead sea bream (Sparus aurata) gills.

It is known that heavy metals have toxic effects on fish. Insufficient measures are a serious problem in our country and around the world. This problem can threaten human health in areas where it is common for people to obtain nutrition from local bodies of water. In this study, the toxicological impacts of some heavy metals were investigated on carbonic anhydrase activity in gilthead gills. Carbonic anhydrase (CA) was purified from gilthead sea bream (Sparus aurata) gills with a specific activity of 2872.92 EU mg(-1) and a yield of 32.84% using affinity chromatography. The overall purification was approximately ∼ 84-fold. SDS-polyacrylamide gel electrophoresis showed a single band, and the MW was approximately 30.5 kDa (Soyut et al., 2008, 2012; Soyut and Beydemir, 2008, 2012; Kaya et al., 2013). The kinetic and characteristic properties of CA such as the optimum pH, stable pH, optimum temperature, activation energy (Ea), activation enthalpy (ΔH), Q10, Km and Vmax were determined. Cadmium (Cd(2+)), copper (Cu(2+)), nickel (Ni(2+)) and silver (Ag(+)) inhibited CA activity in in vitro conditions. Ki values were calculated for these metals. Ki values were 31.20mM for cadmium (Cd(2+)), 161.96 mM for copper (Cu(2+)), 10.79 mM for nickel (Ni(2+)) and 0.0082 mM for silver (Ag(+)) based on Lineweaver-Burk plots. Except for cadmium, heavy metals had the same inhibition mechanism. Cadmium was competitive, and the others were noncompetitive.

Impact of antibacterial drugs on human serum paraoxonase-1 (hPON1) activity: an in vitro study.

OBJECTIVE: To investigate the in vitro effects of the antibacterial drugs, meropenem trihydrate, piperacillin sodium, and cefoperazone sodium, on the activity of human serum paraoxonase (hPON1). METHODS: hPON1 was purified from human serum using simple chromatographic methods, including DEAE-Sephadex anion exchange and Sephadex G-200 gel filtration chromatography. RESULTS: The three antibacterial drugs decreased in vitro hPON1 activity. Inhibition mechanisms meropenem trihydrate was noncompetitive while piperacillin sodium and cefoperazone sodium were competitive. CONCLUSIONS: Our results showed that antibacterial drugs significantly inhibit hPON1 activity, both in vitro, with rank order meropenem trihydrate piperacillin sodium cefoperazone sodium in vitro.

Carbonic anhydrase activity from the gilthead sea bream (Sparus aurata) liver: the toxicological effects of heavy metals.

Many studies have shown that metal ions may lead to oxidative stress in biological systems. Accordingly, DNA damage, protein modification, enzyme inhibition and activation, lipid peroxidation and many other effects may occur in living organisms. Many different formations of metal ions may enter human cells along with water, air, and various foods, and humans are negatively affected by these conditions, either directly or indirectly. These effects may cause irreversible damage to human metabolism. In this study, the toxicological effects of heavy metals on carbonic anhydrase enzyme activity from the gilthead sea bream liver were investigated. The carbonic anhydrase enzyme was purified via affinity chromatography and had a specific activity of 6775.5EUmg(-1). The kinetics and characteristic properties, such as optimum pH, stable pH, optimum temperature, activation energy (Ea), activation enthalpy (ΔH), Q10, Km, and Vmax, were determined for the purified enzyme SDS-polyacrylamide gel electrophoresis showed a single band and molecular weight of the subunit was approximately 25kDa. Cd(II), Cu(II), Ni(II) and Ag(I) inhibited the enzyme activity in vitro. The type of inhibition and Ki values for these metals were calculated from Lineweaver-Burk plots as 17.74mM, 36.20mM, 12.85mM and 0.025mM for Cd(II), Cu(II), Ni(II) and Ag(I), respectively. All the metals were noncompetitive inhibitors.