scholarly journals Further thermal characterization of an aspartate aminotransferase from a halophilic organism

1994 ◽  
Vol 298 (2) ◽  
pp. 465-470 ◽  
Author(s):  
F J G Muriana ◽  
M C Alvarez-Ossorio ◽  
A M Relimpio

Aspartate aminotransferase (AspAT, EC 2.6.1.1) from the halophilic archaebacterium Haloferax mediterranei was purified [Muriana, Alvarez-Ossorio and Relimpio (1991) Biochem. J. 278, 149-154] and further characterization of the effects of temperature on the activity and stability of the halophilic AspAT were carried out. The halophilic transaminase is most active at 65 degrees C and stable at high temperatures, under physiological or nearly physiological conditions (3.5 M KCl, pH 7.8). Thermal inactivation (60-85 degrees C) of the halophilic AspAT followed first-order kinetics, 2-oxoglutarate causing a shift of the thermal inactivation curves to higher temperatures. The salt concentration affected the thermal stability of the halophilic transaminase at 60 degrees C, suggesting that disruption of hydrophobic interactions may play an important role in the decreased thermal stability of the enzyme.

1991 ◽  
Vol 278 (1) ◽  
pp. 149-154 ◽  
Author(s):  
F J G Muriana ◽  
M C Alvarez-Ossorio ◽  
A M Relimpio

Aspartate aminotransferase from the archaebacterium Haloferax mediterranei was purified and found to be homogeneous. An average Mr of 66,000 was estimated. The native halophilic transaminase exhibited no maximum absorption at 410 nm, which indicates that the apo form is obtained by our purification procedure, and the molar absorption coefficient at 275 nm in 3.5 M-KCl (pH 7.8) was found to be 78.34 mM-1.cm-1. Plots of titration data show that 1 mol of halophilic aspartate aminotransferase binds 2 mol of pyridoxal 5′-phosphate. The halophilic transaminase behaved as a dimer with two similar subunits and had a maximum activity in the pH range 7.6-7.9 and at 65 degrees C in 3.5 M-KCl. By differential scanning calorimetry, the denaturation temperature of the halophilic holo- and apo-transaminase was determined to be 78.5 and 68.0 degrees C respectively at 3.3 M-KCl (pH 7.8). At low salt concentration the halophilic transaminase was inactivated, following first-order kinetics. The Km values for 2-oxoglutarate and L-aspartate, in 3 M-KCl (pH 7.8), were 0.75 mM and 12.6 mM respectively.


1993 ◽  
Vol 295 (3) ◽  
pp. 827-831 ◽  
Author(s):  
A Villa ◽  
L Zecca ◽  
P Fusi ◽  
S Colombo ◽  
G Tedeschi ◽  
...  

Investigations were performed on the structural features responsible for kinetic thermal stability of a thermostable carboxypeptidase from the thermoacidophilic archaebacterium Sulfolobus solfataricus which had been purified previously and identified as a zinc metalloprotease [Colombo, D'Auria, Fusi, Zecca, Raia and Tortora (1992) Eur. J. Biochem. 206, 349-357]. Removal of Zn2+ by dialysis led to reversible activity loss, which was promptly restored by addition of 80 microM ZnCl2 to the assay mixture. For the first-order irreversible thermal inactivation the metal-depleted enzyme showed an activation energy value of 205.6 kJ.mol-1, which is considerably lower than that of the holoenzyme (494.4 kJ.mol-1). The values of activation free energies, enthalpies and entropies also dropped with metal removal. Thermal inactivation of the apoenzyme was very quick at 80 degrees C, whereas the holoenzyme was stable at the same temperature. These findings suggest a major stabilizing role for the bivalent cation. Chaotropic salts strongly destabilized the holoenzyme, showing that hydrophobic interactions are involved in maintaining the native conformation of the enzyme. However, the inactivation rate was also increased by sodium sulphate, acetate and chloride, which are not chaotropes, indicating that one or more salt bridges concur in stabilizing the active enzyme. Furthermore, at the extremes of the pH-stability curve, NaCl did not affect the inactivation rate, confirming the stabilizing role of intramolecular ionic bonds, as a pH-dependent decrease in stability is likely to occur from breaking of salt bridges involved in maintaining the native conformation of the protein.


Author(s):  
Bernard Téhi Sea ◽  
Armand Kouassi Kanga ◽  
Djary Michel Koffi ◽  
Lucien Patrice Kouamé

Polygalacturonases are extensively used in food industries for pectic substances degradation. In this paper, we investigate on thermal stability parameters of two Polygalacturonases previously isolated from digestive juice of the snail Limicolaria flammea for several industrial applications such as fruit juice clarification. Thermal inactivation was carried out in the temperature range of 55°C to 80°C from 15 to 120 min. All results were statistically analysed. The results shown that thermal inactivation of studied acid phosphatases follows first order kinetics. At their optimum temperatures, these enzymes showed high half-lives ranging from 462.06 to 630.10 min and D values from 1535.00 to 2093.64 min suggesting that these two enzymes had a large thermal stability. The high values of ΔG# (93.96 to 94.97 kJ/mol) reveal a better resistance to denaturation. The relatively high activation energies (from 120.35 to 129.13 kJ/mol) and average enthalpy values (from 117.67 to 126.44 kJ.mol−1) could corroborate the good stability of these biocatalyst. All these results suggest that Polygalacturonases from digestive juice of the snail Limicolaria flammea may be profitably exploited in future food industrial applications.


2019 ◽  
pp. 089270571987822
Author(s):  
Saud Aldajah ◽  
Mohammad Y Al-Haik ◽  
Waseem Siddique ◽  
Mohammad M Kabir ◽  
Yousef Haik

This study reveals the enhancement of mechanical and thermal properties of maleic anhydride-grafted polypropylene (PP- g-MA) with the addition of nanocrystalline cellulose (NCC). A nanocomposite was manufactured by blending various percentages of PP, MA, and NCC nanoparticles by means of a twin-screw extruder. The influence of varying the percentages of NCC on the mechanical and thermal behavior of the nanocomposite was studied by performing three-point bending, nanoindentation, differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), scanning electron microscopy (SEM), and Fourier-transform infrared (FTIR) spectroscopy tests. The novelty of this study stems on the NCC nanoparticles and their ability to enhance the mechanical and thermal properties of PP. Three-point bending and nanoindentation tests revealed improvement in the mechanical properties in terms of strength, modulus, and hardness of the PP- g-MA nanocomposites as the addition of NCC increased. SEM showed homogeneity between the mixtures which proved the presence of interfacial adhesion between the PP- g-MA incorporated with NCC nanoparticles that was confirmed by the FTIR results. DSC and TGA measurements showed that the thermal stability of the nanocomposites was not compromised due to the addition of the coupling agent and reinforced nanoparticles.


2021 ◽  
Author(s):  
Qifeng Jiang ◽  
Sydnee Wong ◽  
Rebekka S Klausen

Thermal characterization of polysilanes has focused on the influence of organic side chains, whereas little is understood about the influence of silane backbone microstructure on thermal stability, phase properties, and...


e-Polymers ◽  
2012 ◽  
Vol 12 (1) ◽  
Author(s):  
Benhong Yang ◽  
Meng Li ◽  
Yun Wu ◽  
Kang Wang

AbstractSeveral inorganic/organic nanocomposites were prepared via solution-blending of cage-like octahexyl-polyhedral oligomeric silsesquioxane (Oh-POSS) with polystyrene (PS) in THF solvent. FTIR and 29Si-NMR were employed to characterize the structures of the nanocomposites. SEM pictures showed that the sample films were smooth and no POSS aggregation was observed when POSS content was lower than 1.0 wt%. TGA and DSC were used to investigate the thermal property. The results showed that the incorporation of nanosized Oh-POSS enhanced the thermal stability of PS with low POSS content. When 1.0 wt% of Oh-POSS was incorporated into PS matrix, the Tg and Td increased by 7.7 °C and 8.2 °C, respectively. However, higher POSS contents (>1.0 wt%) would deteriorate the thermal property of the nanocomposites due to the severe congregation of POSS..


2012 ◽  
Vol 727-728 ◽  
pp. 1552-1556
Author(s):  
Renata Barbosa ◽  
Dayanne Diniz Souza ◽  
Edcleide Maria Araújo ◽  
Tomás Jefférson Alves de Mélo

Studies of degradation have verified that the decomposition of some quaternary ammonium salts can begin to be significant at the temperature of about 180 ° C and like most thermoplastics are processed at least around this temperature, the thermal stability of the salt in clay should always be considered. Some salts are more stable than others, being necessary to study the degradation mechanisms of each case. In this work, four quaternary ammonium salts were characterized by differential scanning calorimetry (DSC) and thermogravimetry (TG). The results of DSC and TG showed that the salts based chloride (Cl-) anion begin to degrade at similar temperatures, while the salt based bromide (Br-) anion degrades at higher temperature. Subsequently, a quaternary ammonium salt was chosen to be used in organoclays, depending on its chemical structure and its thermal behavior.


2001 ◽  
Vol 288 (1) ◽  
pp. 121-128 ◽  
Author(s):  
Serguei V. Kuznetsov ◽  
Olga M. Sidorkina ◽  
Jacques Laval ◽  
Anjum Ansari

2011 ◽  
Vol 197-198 ◽  
pp. 606-609 ◽  
Author(s):  
Ti Feng Jiao ◽  
Yuan Yuan Xing ◽  
Jing Xin Zhou ◽  
Wei Wang

Some functional luminol derivatives with aromatic substituted groups have been designed and synthesized from the reaction of the corresponding aromatic acyl chloride precursors with luminol. It has been found that depending on the size of aromatic groups, the formed luminol derivatives showed different properties, indicating distinct regulation of molecular skeletons. UV and IR data confirmed commonly the formation of imide group as well as aromatic segment in molecular structures. Thermal analysis showed that the thermal stability of luminol derivatives with p-phthaloyl segment was the highest in those derivatives. The difference of thermal stability is mainly attributed to the formation of imide group and aromatic substituent groups in molecular structure. The present results have demonstrated that the special properties of luminol derivatives can be turned by modifying molecular structures of objective compounds with proper substituted groups, which show potential application in functional material field and ECL sensor.


Author(s):  
Mahbubor Rahman ◽  
Tuba DEMIREL ◽  
Ismail KARACAN

Thermal oxidation of polyacrylonitrile (PAN) fibers was accomplished at temperatures up to 250°C for different oxidation times. Chemical integration of PAN fibers with an aqueous solution of ammonium persulfate was performed before starting thermal oxidation. The results recommend that ammonium persulfate integration enhanced the oxidation reactions of the PAN fibers and resulting in enhanced thermal stability. Ammonium persulfate impregnation followed by the oxidation process in the air environment leads to significant deviations in the characteristics of PAN fibers. To perform structural characterization of the raw and ammonium persulfate (APS) incorporated and stabilized samples, XRD, IR-spectroscopy, TGA was executed in this study. Investigation of the XRD and infrared spectroscopy outcomes recommended quick aromatization reactions with growing oxidation periods. The TGA traces indicated a comparative enhancement in the thermal stability of the PAN fibers by the increased carbon yield with the rise of the oxidation time. The overall findings recommend that ammonium persulfate incorporation was very influential in stimulating the oxidation process.


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