scholarly journals Immobilization of Mutant Phosphotriesterase on Fuller’s Earth Enhanced the Stability of the Enzyme

Catalysts ◽  
2021 ◽  
Vol 11 (8) ◽  
pp. 983
Author(s):  
Wahhida Latip ◽  
Victor Feizal Knight ◽  
Ong Keat Khim ◽  
Noor Azilah Mohd Kasim ◽  
Wan Md Zin Wan Yunus ◽  
...  
Keyword(s):  
Thermal Stability ◽  
Metal Ions ◽  
Half Life ◽  
Industrial Applications ◽  
Free Enzyme ◽  
High Tolerance ◽  
Tween 40 ◽  
Fuller’S Earth ◽  
The Stability ◽  
Biochemical Features

Immobilization is a method for making an enzyme more robust in the environment, especially in terms of its stability and reusability. A mutant phosphotriesterase (YT PTE) isolated from Pseudomonas dimunita has been reported to have high proficiency in hydrolyzing the Sp and Rp-enantiomers of organophosphate chromophoric analogs and therefore has great potential as a decontamination agent and biosensor. This work aims to investigate the feasibility of using Fuller’s earth (FE) as a YT PTE immobilization support and characterize its biochemical features after immobilization. The immobilized YT PTE was found to show improvement in thermal stability with a half-life of 24 h compared to that of the free enzyme, which was only 8 h. The stability of the immobilized YT PTE allowed storage for up to 4 months and reuse for up to 6 times. The immobilized YT PTE showed high tolerance against all tested metal ions, Tween 40 and 80 surfactants and inorganic solvents. These findings showed that the immobilized YT PTE became more robust for use especially with regards to its stability and reusability. These features would enhance the future applicability of this enzyme as a decontamination agent and its use in other suitable industrial applications.

scholarly journals Lysosomal protein thermal stability does not correlate with cellular half-life: global observations and a case study of tripeptidyl-peptidase 1

2020 ◽  
Vol 477 (3) ◽  
pp. 727-745 ◽  
Author(s):  
Aaron M. Collier ◽  
Yuliya Nemtsova ◽  
Narendra Kuber ◽  
Whitney Banach-Petrosky ◽  
Anurag Modak ◽  
...  
Keyword(s):  
Thermal Stability ◽  
Protein Engineering ◽  
Half Life ◽  
Neuronal Ceroid Lipofuscinosis ◽  
Turnover Rates ◽  
Lysosomal Storage ◽  
Enzyme Replacement ◽  
Tripeptidyl Peptidase ◽  
The Stability

Late-infantile neuronal ceroid lipofuscinosis (LINCL) is a neurodegenerative lysosomal storage disorder caused by mutations in the gene encoding the protease tripeptidyl-peptidase 1 (TPP1). Progression of LINCL can be slowed or halted by enzyme replacement therapy, where recombinant human TPP1 is administered to patients. In this study, we utilized protein engineering techniques to increase the stability of recombinant TPP1 with the rationale that this may lengthen its lysosomal half-life, potentially increasing the potency of the therapeutic protein. Utilizing multiple structure-based methods that have been shown to increase the stability of other proteins, we have generated and evaluated over 70 TPP1 variants. The most effective mutation, R465G, increased the melting temperature of TPP1 from 55.6°C to 64.4°C and increased its enzymatic half-life at 60°C from 5.4 min to 21.9 min. However, the intracellular half-life of R465G and all other variants tested in cultured LINCL patient-derived lymphoblasts was similar to that of WT TPP1. These results provide structure/function insights into TPP1 and indicate that improving in vitro thermal stability alone is insufficient to generate TPP1 variants with improved physiological stability. This conclusion is supported by a proteome-wide analysis that indicates that lysosomal proteins have higher melting temperatures but also higher turnover rates than proteins of other organelles. These results have implications for similar efforts where protein engineering approaches, which are frequently evaluated in vitro, may be considered for improving the physiological properties of proteins, particularly those that function in the lysosomal environment.

scholarly journals Protein thermal stability does not correlate with cellular half-life: Global observations and a case study of tripeptidyl-peptidase 1

2019 ◽  
Author(s):  
Aaron M. Collier ◽  
Yuliya Nemtsova ◽  
Narendra Kuber ◽  
Whitney Banach-Petrosky ◽  
Anurag Modak ◽  
...  
Keyword(s):  
Thermal Stability ◽  
Protein Engineering ◽  
Half Life ◽  
Neuronal Ceroid Lipofuscinosis ◽  
Turnover Rates ◽  
Lysosomal Storage ◽  
Enzyme Replacement ◽  
Tripeptidyl Peptidase ◽  
The Stability

AbstractLate-infantile neuronal ceroid lipofuscinosis (LINCL) is a neurodegenerative lysosomal storage disorder caused by mutations in the gene encoding the protease tripeptidyl-peptidase 1 (TPP1). Progression of LINCL can be slowed or halted by enzyme replacement therapy, where recombinant human TPP1 is administered to patients. In this study, we utilized protein engineering techniques to increase the stability of recombinant TPP1 with the rationale that this may lengthen its lysosomal half-life, potentially increasing the potency of the therapeutic protein. Utilizing multiple structure-based methods that have been shown to increase the stability of other proteins, we have generated and evaluated over 70 TPP1 variants. The most effective mutation, R465G, increased the melting temperature of TPP1 from 55.6°C to 64.4°C and increased its enzymatic half-life at 60°C from 5.4 min to 21.9 min. However, the intracellular half-life of R465G and all other variants tested in cultured LINCL-patient derived lymphoblasts was similar to that of WT TPP1. These results provide structure/function insights into TPP1 and indicate that improving in vitro thermal stability alone is insufficient to generate TPP1 variants with improved physiological stability. This conclusion is supported by a proteome-wide analysis that indicates that lysosomal proteins have higher melting temperatures but also higher turnover rates than proteins of other organelles. These results have implications for similar efforts where protein engineering approaches, which are frequently evaluated in vitro, may be considered for improving the physiological properties of proteins, particularly those that function in the lysosomal environment.

Kinetics of Various 99mTc-Sn-Pyrophosphate Compounds in the Rat

1977 ◽  
Vol 16 (04) ◽  
pp. 157-162 ◽  
Author(s):  
C. Schümichen ◽  
B. Mackenbrock ◽  
G. Hoffmann
Keyword(s):  
Normal Saline ◽  
Half Life ◽  
Compound A ◽  
Short Half Life ◽  
Low Concentrations ◽  
The Stability ◽  
Kinetics Of ◽  
In Vitro Stability

SummaryThe bone-seeking 99mTc-Sn-pyrophosphate compound (compound A) was diluted both in vitro and in vivo and proved to be unstable both in vitro and in vivo. However, stability was much better in vivo than in vitro and thus the in vitro stability of compound A after dilution in various mediums could be followed up by a consecutive evaluation of the in vivo distribution in the rat. After dilution in neutral normal saline compound A is metastable and after a short half-life it is transformed into the other 99mTc-Sn-pyrophosphate compound A is metastable and after a short half-life in bone but in the kidneys. After dilution in normal saline of low pH and in buffering solutions the stability of compound A is increased. In human plasma compound A is relatively stable but not in plasma water. When compound B is formed in a buffering solution, uptake in the kidneys and excretion in urine is lowered and blood concentration increased.It is assumed that the association of protons to compound A will increase its stability at low concentrations while that to compound B will lead to a strong protein bond in plasma. It is concluded that compound A will not be stable in vivo because of a lack of stability in the extravascular space, and that the protein bond in plasma will be a measure of its in vivo stability.

REMOVAL OF HEAVY METAL Cr (II), Ni (II), Cu (II), Zn (II), Cd (II), Pb (II) IONS FROM AQUEOUS SOLUTION BY MENTHA PIPERITA EXTRACT

2020 ◽  
pp. 15-20
Author(s):  
Ersin Yucel ◽  
Mine Yucel
Keyword(s):  
Aqueous Solution ◽  
Metal Ions ◽  
High Efficiency ◽  
Correlation Coefficients ◽  
Langmuir Model ◽  
Mentha Piperita ◽  
Freundlich Model ◽  
Biosorption Capacity ◽  
Peppermint Extract ◽  
The Stability

In this study, the usage of the peppermint (Mentha piperita) for extracting the metal ions [Mg (II), Cr (II), Ni (II), Cu (II), Zn (II), Cd (II), Pb (II)] that exist at water was investigated. In order to analyze the stability properties, Langmuir, Freundlich, Temkin and Dubinin-Radushkevich isotherms were used at removing the metal ions and the highest correlation coefficients (R2) were obtained at Langmuir isotherm. Therefore, it is seen that the Langmuir model is more proper than the Freundlich model. However, it was found that the correlation coefficients of removing Ni and Cd is higher at Freundlich model than Langmuir and low at Dubinin-Radushkevich isotherm. It is established that the biosorption amount increase depends on the increase of biosorbent and it can be achieved high efficiency (95%) even with small amount (0.6 mg, peppermint extract) at lead ions. It is also determined that the peppermint extracted that is used at this study shows high biosorption capacity for metal ions and can be used for immobilization of metals from polluted areas.

Optimization of Organic Fibres%[Kevlar/Arobocel/Acrylic] in NAO Brake Pad Application and its Effect on Thermal Stability & Friction Characteristics

2012 ◽  
Vol 531-532 ◽  
pp. 8-12
Author(s):  
M.A. Sai Balaji ◽  
K. Kalaichelvan
Keyword(s):  
Thermal Stability ◽  
Weight Loss ◽  
Minimum Weight ◽  
Friction Material ◽  
Brake Pad ◽  
Temperature Zone ◽  
Good Thermal Stability ◽  
Friction Testing ◽  
Brake Application ◽  
The Stability

Organic fibres (Kevlar/ Arbocel / Acrylic) have good thermal stability, higher surface area and bulk density. The optimization of organic fibres percentage for thermal behaviour is considered using TGA. The temperature raise during brake application will be between 150-4000 C and this temperature zone is very critical to determine the fade characteristics during friction testing. Hence, three different friction composites are developed with the same formulation varying only the Kevlar, Arbocel and Acrylic fibres which are compensated by the inert filler namely the barites and are designated as NA01, NA02 and NA03 respectively. After the fabrication, the TGA test reveals that the composite NA03 has minimum weight loss. The friction coefficient test rig is then used to test the friction material as per SAE J661a standards. The results prove that the brake pad with minimum weight loss during TGA has higher friction stability. Thus, we can correlate the thermal stability with the stability of friction.

scholarly journals Investigation of the Stability of Yeast rad52 Mutant Proteins Uncovers Post-translational and Transcriptional Regulation of Rad52p

Genetics ◽  
2003 ◽  
Vol 163 (1) ◽  
pp. 91-101 ◽  
Author(s):  
Erin N Asleson ◽  
Dennis M Livingston
Keyword(s):  
Half Life ◽  
Translational Regulation ◽  
Cellular Level ◽  
Missense Mutations ◽  
Wild Type ◽  
Mutant Proteins ◽  
Gal1 Promoter ◽  
Rad52 Protein ◽  
The Stability ◽  
Mutant Forms

Abstract We investigated the stability of the Saccharomyces cerevisiae Rad52 protein to learn how a cell controls its quantity and longevity. We measured the cellular levels of wild-type and mutant forms of Rad52p when expressed from the RAD52 promoter and the half-lives of the various forms of Rad52p when expressed from the GAL1 promoter. The wild-type protein has a half-life of 15 min. rad52 mutations variably affect the cellular levels of the protein products, and these levels correlate with the measured half-lives. While missense mutations in the N terminus of the protein drastically reduce the cellular levels of the mutant proteins, two mutations—one a deletion of amino acids 210-327 and the other a missense mutation of residue 235—increase the cellular level and half-life more than twofold. These results suggest that Rad52p is subject to post-translational regulation. Proteasomal mutations have no effect on Rad52p half-life but increase the amount of RAD52 message. In contrast to Rad52p, the half-life of Rad51p is >2 hr, and RAD51 expression is unaffected by proteasomal mutations. These differences between Rad52p and Rad51p suggest differential regulation of two proteins that interact in recombinational repair.

scholarly journals Suboptimal Disturbance Observer Design Using All Stabilizing Q Filter for Precise Tracking Control

Mathematics ◽  
2020 ◽  
Vol 8 (9) ◽  
pp. 1434 ◽  
Author(s):  
Wonhee Kim ◽  
Sangmin Suh
Keyword(s):  
Design Method ◽  
Industrial Applications ◽  
Point Of View ◽  
Observer Design ◽  
Linear Matrix ◽  
External Disturbances ◽  
Closed Loop Systems ◽  
Control Target ◽  
The Stability ◽  
Unified Index

For several decades, disturbance observers (DOs) have been widely utilized to enhance tracking performance by reducing external disturbances in different industrial applications. However, although a DO is a verified control structure, a conventional DO does not guarantee stability. This paper proposes a stability-guaranteed design method, while maintaining the DO structure. The proposed design method uses a linear matrix inequality (LMI)-based H∞ control because the LMI-based control guarantees the stability of closed loop systems. However, applying the DO design to the LMI framework is not trivial because there are two control targets, whereas the standard LMI stabilizes a single control target. In this study, the problem is first resolved by building a single fictitious model because the two models are serial and can be considered as a single model from the Q-filter point of view. Using the proposed design framework, all-stabilizing Q filters are calculated. In addition, for the stability and robustness of the DO, two metrics are proposed to quantify the stability and robustness and combined into a single unified index to satisfy both metrics. Based on an application example, it is verified that the proposed method is effective, with a performance improvement of 10.8%.

Photoinhibition of Electron Transport Activity of Photosystem II in Isolated Thylakoids Studied by Thermoluminescence and Delayed Luminescence

1988 ◽  
Vol 43 (11-12) ◽  
pp. 871-876 ◽  
Author(s):  
Imre Vass ◽  
Narendranath Mohanty ◽  
Sándor Demeter
Keyword(s):  
Electron Transport ◽  
Photosystem Ii ◽  
Photosystem I ◽  
Half Life ◽  
Delayed Luminescence ◽  
Transport Activity ◽  
Primary Target ◽  
Electron Transport Activity ◽  
The Stability ◽  
Spinach Thylakoids

Abstract The effect of photoinhibition on the primary (QA) and secondary (QB) quinone acceptors of photosystem I I was investigated in isolated spinach thylakoids by the methods of thermoluminescence and delayed luminescence. The amplitudes of the Q (at about 2 °C) and B (at about 30 °C) thermoluminescence bands which are associated with the recombination of the S2QA- and S2QB charge pairs, respectively, exhibited parallel decay courses during photoinhibitory treatment. Similarly, the amplitudes of the flash-induced delayed luminescence components ascribed to the recombination of S20A and S2OB charge pairs and having half life-times of about 3 s and 30 s, respectively, declined in parallel with the amplitudes of the corresponding Q and B thermoluminescence bands. The course of inhibition of thermoluminescence and delayed luminescence intensity was parallel with that of the rate of oxygen evolution. The peak positions of the B and Q thermoluminescence bands as well as the half life-times of the corresponding delayed luminescence components were not affected by photoinhibition. These results indicate that in isolated thylakoids neither the amount nor the stability of the reduced OB acceptor is preferentially decreased by photoinhibition. We conclude that either the primary target of photodamage is located before the O b binding site in the reaction center of photosystem II or QA and OB undergo simultaneous damage.

A DENSITY FUNCTIONAL STUDY OF THE STRUCTURE OF SILANE COUPLING AGENT 3–AMINOPROPYLTRIETHOXYSILANE

2005 ◽  
Vol 04 (01) ◽  
pp. 117-126
Author(s):  
N. L. MA ◽  
P. WU
Keyword(s):  
Coupling Agent ◽  
Density Functional ◽  
Solution Structure ◽  
Silane Coupling Agent ◽  
Industrial Applications ◽  
Functional Study ◽  
Stable Form ◽  
Functional Theory ◽  
Silane Coupling ◽  
The Stability

Using density functional theory, we predicted the solution structure of the hydrolyzed 3–aminopropyltriethoxysilane (h–APS), which is a silane coupling agent commonly used in many industrial applications. We have located five stable minima on the potential energy surface of h–APS in which four of them are "neutral", and the remaining one is zwitterionic (dipolar) in nature. Our calculations suggested that the stability of the most stable form of h–APS in water (denoted as II_N) arose from strong intramolecular OH ⋯ N hydrogen bond. The least stable form is the zwitterionic form (I_ZW), which is estimated to be over 90 kJ mol -1 less stable than II_N. The factors governing the relative stabilities of different forms are discussed.

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