scholarly journals Thermostable Adenosine 5’-Monophosphate Phosphorylase from Thermococcus kodakarensis forms catalytically active inclusion bodies

2020 ◽  
Author(s):  
Sarah Kamel ◽  
Miriam Walczak ◽  
Felix Kaspar ◽  
Sarah Westarp ◽  
Peter Neubauer ◽  
...  
Keyword(s):  
Heat Treatment ◽  
Thermal Stability ◽  
Inclusion Bodies ◽  
Specific Activity ◽  
High Thermal Stability ◽  
Active Protein ◽  
E Coli ◽  
Thermococcus Kodakarensis ◽  
Active Inclusion Bodies ◽  
Catalytically Active

Abstract Catalytically active inclusion bodies (CatIBs) produced in E. coli are an interesting but currently underexplored strategy for enzyme immobilization. They can be purified easily and used directly as stable and reusable heterogenous catalysts. However, very few examples of CatIBs that are naturally formed during heterologous expression have been reported so far. Previous studies have revealed that the adenosine 5'-monophosphate phosphorylase of Thermococcus kodakarensis (TkAMPpase) forms large soluble multimers with high thermal stability. Herein, we show that heat treatment of solubilized protein induces aggregation of active protein which phosphorolysis all natural 5’-mononucleotides. Additionally, inclusion bodies formed during the expression in E. coli were found to be similarly active with 2−6 folds higher specific activity compared to the heat-induced aggregates. Interestingly, differences in the substrate preference were observed. These results show that the recombinant thermostable TkAMPpase is one of rare examples of naturally formed CatIBs.

scholarly journals Thermostable adenosine 5′-monophosphate phosphorylase from Thermococcus kodakarensis forms catalytically active inclusion bodies

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Sarah Kamel ◽  
Miriam C. Walczak ◽  
Felix Kaspar ◽  
Sarah Westarp ◽  
Peter Neubauer ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Heat Treatment ◽  
Inclusion Bodies ◽  
Specific Activity ◽  
High Thermal Stability ◽  
Active Protein ◽  
E Coli ◽  
Thermococcus Kodakarensis ◽  
Active Inclusion Bodies ◽  
Catalytically Active

AbstractCatalytically active inclusion bodies (CatIBs) produced in Escherichia coli are an interesting but currently underexplored strategy for enzyme immobilization. They can be purified easily and used directly as stable and reusable heterogenous catalysts. However, very few examples of CatIBs that are naturally formed during heterologous expression have been reported so far. Previous studies have revealed that the adenosine 5′-monophosphate phosphorylase of Thermococcus kodakarensis (TkAMPpase) forms large soluble multimers with high thermal stability. Herein, we show that heat treatment of soluble protein from crude extract induces aggregation of active protein which phosphorolyse all natural 5′-mononucleotides. Additionally, inclusion bodies formed during the expression in E. coli were found to be similarly active with 2–6 folds higher specific activity compared to these heat-induced aggregates. Interestingly, differences in the substrate preference were observed. These results show that the recombinant thermostable TkAMPpase is one of rare examples of naturally formed CatIBs.

scholarly journals Catalytically active inclusion bodies of L-lysine decarboxylase from E. coli for 1,5-diaminopentane production

2018 ◽  
Vol 8 (1) ◽  
Author(s):  
Ramona Kloss ◽  
Michael H. Limberg ◽  
Ursula Mackfeld ◽  
Doris Hahn ◽  
Alexander Grünberger ◽  
...  
Keyword(s):  
Inclusion Bodies ◽  
Lysine Decarboxylase ◽  
E Coli ◽  
Active Inclusion Bodies ◽  
Catalytically Active

Binding of 3-phosphoglycerate leads to both activation and stabilisation of ADP-glucose pyrophosphorylase from apple leaves

2005 ◽  
Vol 32 (9) ◽  
pp. 839
Author(s):  
Rui Zhou ◽  
Lailiang Cheng
Keyword(s):  
Heat Treatment ◽  
Thermal Stability ◽  
Enzyme Activity ◽  
Inorganic Phosphate ◽  
Thermal Inactivation ◽  
Specific Activity ◽  
Apple Leaves ◽  
Apparent Homogeneity ◽  
Adp Glucose Pyrophosphorylase ◽  
High Concentrations

Apple leaf ADP-glucose pyrophosphorylase was purified 1436-fold to apparent homogeneity with a specific activity of 58.9 units mg–1. The enzyme was activated by 3-phosphoglycerate (PGA) and inhibited by inorganic phosphate (Pi) in the ADPG synthesis direction. In the pyrophosphorolytic direction, however, high concentrations of PGA (> 2.5 mm) inhibited the enzyme activity. The enzyme was resistant to thermal inactivation with a T0.5 (temperature at which 50% of the enzyme activity is lost after 5 min incubation) of 52°C. Incubation with 2 mm PGA or 2 mm Pi increased T0.5 to 68°C. Incubation with 2 mm dithiothreitol (DTT) decreased T0.5 to 42°C, whereas inclusion of 2 mm PGA in the DTT incubation maintained T0.5 at 52°C. DTT-induced decrease in thermal stability was accompanied by monomerisation of the small subunits. Presence of PGA in the DTT incubation did not alter the monomerisation of the small subunits of the enzyme induced by DTT. These findings indicate that binding of PGA renders apple leaf AGPase with a conformation that is not only more efficient in catalysis but also more stable to heat treatment. The physiological significance of the protective effect of PGA on thermal inactivation of AGPase is discussed.

Catalytically-active inclusion bodies—Carrier-free protein immobilizates for application in biotechnology and biomedicine

2017 ◽  
Vol 258 ◽  
pp. 136-147 ◽  
Author(s):  
Ulrich Krauss ◽  
Vera D. Jäger ◽  
Martin Diener ◽  
Martina Pohl ◽  
Karl-Erich Jaeger
Keyword(s):  
Inclusion Bodies ◽  
Free Protein ◽  
Active Inclusion Bodies ◽  
Carrier Free ◽  
Catalytically Active

SpyTag/Catcher chemistry induces the formation of active inclusion bodies in E. coli

2022 ◽  
Author(s):  
Wenge Dong ◽  
Hongxu Sun ◽  
Qiwei Chen ◽  
Liangyu Hou ◽  
Yanhong Chang ◽  
...  
Keyword(s):  
Inclusion Bodies ◽  
E Coli ◽  
Active Inclusion Bodies

scholarly journals Catalytically-active inclusion bodies for biotechnology—general concepts, optimization, and application

2020 ◽  
Vol 104 (17) ◽  
pp. 7313-7329
Author(s):  
Vera D. Jäger ◽  
Robin Lamm ◽  
Kira Küsters ◽  
Gizem Ölçücü ◽  
Marco Oldiges ◽  
...  
Keyword(s):  
Inclusion Bodies ◽  
Active Inclusion Bodies ◽  
Catalytically Active

TOUGHMET 2

Alloy Digest ◽  
2013 ◽  
Vol 62 (5) ◽  
Keyword(s):  
Heat Treatment ◽  
Thermal Stability ◽  
Fracture Toughness ◽  
Wear Resistance ◽  
High Performance ◽  
Mechanical Performance ◽  
High Thermal Stability ◽  
Lead Free ◽  
Plain Bearing ◽  
Ni Alloy

Abstract ToughMet 2 is a high performance, wrought, heat treatable, lead-free strip Cu-Ni alloy that imparts superior mechanical performance and high thermal stability to plain bearing applications. Parts are easily formed and they can be machined either before or after heat treatment. ToughMet alloys are a line of spinodal hardened Cu-Ni anti-galling alloys for bearings capable of performing with a variety of shafting materials and lubricants. The alloys combine a high lubricity with wear resistance in these severe loading conditions. This datasheet provides information on composition, physical properties, hardness, elasticity, and tensile properties as well as fracture toughness and fatigue. It also includes information on corrosion resistance as well as forming and machining. Filing Code: Cu-724. Producer or source: Materion Brush Performance Alloys. Originally published September 2004, revised May 2013.

scholarly journals Tailoring the properties of (catalytically)-active inclusion bodies

2019 ◽  
Vol 18 (1) ◽  
Author(s):  
V. D. Jäger ◽  
R. Kloss ◽  
A. Grünberger ◽  
S. Seide ◽  
D. Hahn ◽  
...  
Keyword(s):  
Inclusion Bodies ◽  
Active Inclusion Bodies ◽  
Catalytically Active
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