Purification and characterization of highly active and stable polyphosphatase fromSaccharomyces cerevisiae cell envelope

Yeast ◽  
1993 ◽  
Vol 9 (2) ◽  
pp. 127-139 ◽  
Author(s):  
Nadezhda A. Andreeva ◽  
Lev A. Okorokov
Keyword(s):  
Cell Envelope ◽  
Purification And Characterization ◽  
Highly Active

scholarly journals Purification and Characterization of the Bacterial UDP-GlcNAc:Undecaprenyl-Phosphate GlcNAc-1-Phosphate Transferase WecA

2008 ◽  
Vol 190 (21) ◽  
pp. 7141-7146 ◽  
Author(s):  
Bayan Al-Dabbagh ◽  
Dominique Mengin-Lecreulx ◽  
Ahmed Bouhss
Keyword(s):  
Cell Envelope ◽  
Functional Characterization ◽  
Integral Membrane Protein ◽  
Minimal Length ◽  
Purification And Characterization ◽  
Effects Of Ph ◽  
Undecaprenyl Phosphate ◽  
First Time ◽  
Lipid Substrate

ABSTRACT To date, the structural and functional characterization of proteins belonging to the polyprenyl-phosphate N-acetylhexosamine-1-phosphate transferase superfamily has been relentlessly held back by problems encountered with their overexpression and purification. In the present work and for the first time, the integral membrane protein WecA that catalyzes the transfer of the GlcNAc-1-phosphate moiety from UDP-GlcNAc onto the carrier lipid undecaprenyl phosphate, yielding undecaprenyl-pyrophosphoryl-GlcNAc, the lipid intermediate involved in the synthesis of various bacterial cell envelope components, was overproduced and purified to near homogeneity in milligram quantities. An enzymatic assay was developed, and the kinetic parameters of WecA as well as the effects of pH, salts, cations, detergents, and temperature on the enzyme activity were determined. A minimal length of 35 carbons was required for the lipid substrate, and tunicamycin was shown to inhibit the enzyme at submicromolar concentrations.

Isolation, purification, and characterization of highly active protease from Bacillus subtilis-150

2010 ◽  
Vol 46 (5) ◽  
pp. 833-834
Author(s):  
K. T. Normurodova ◽  
A. A. Makhsumkhanov ◽  
B. Kh. Alimova ◽  
O. M. Pulatova ◽  
N. I. Bozorov
Keyword(s):  
Bacillus Subtilis ◽  
Purification And Characterization ◽  
Highly Active ◽  
Active Protease

Purification and Characterization of an Alkaline Protease from Bacillus alcalophilus

2012 ◽  
Vol 192 ◽  
pp. 285-288 ◽  
Author(s):  
Shan Shan Liu ◽  
Li Li Wang ◽  
Lin Yuan
Keyword(s):  
Alkaline Protease ◽  
Exchange Chromatography ◽  
Cation Exchange Chromatography ◽  
Ammonium Sulfate Precipitation ◽  
Purification And Characterization ◽  
Highly Active ◽  
Wide Range ◽  
Commercial Applications ◽  
Ph Range

The purification and characterization of an alkaline protease produced by Bacillus alcalophilus were investigated. The enzyme was purified in two steps: concentrating the crude enzyme using ammonium sulfate precipitation, followed by cation-exchange chromatography. The purified protease had a molecular mass of approximately 28 kDa, was highly active over an alkaline pH range of 10.0 to 11.0, and remained stable over a pH range of 7.0 to 12.0. The optimum temperature for the enzyme activity was found to be 40~60°C, while the thermotolerance of the enzyme was poor. Therefore, these characteristics of the protease indicate its potential for a wide range of commercial applications.

scholarly journals Heterologous Expression, Purification, and Characterization of a Highly Active Xylose Reductase from Neurospora crassa

2005 ◽  
Vol 71 (3) ◽  
pp. 1642-1647 ◽  
Author(s):  
Ryan Woodyer ◽  
Michael Simurdiak ◽  
Wilfred A. van der Donk ◽  
Huimin Zhao
Keyword(s):  
Neurospora Crassa ◽  
Xylose Reductase ◽  
Whole Genome Sequence ◽  
High Yield ◽  
In Vitro Production ◽  
Purification And Characterization ◽  
Highly Active ◽  
Vitro Production

ABSTRACT A xylose reductase (XR) gene was identified from the Neurospora crassa whole-genome sequence, expressed heterologously in Escherichia coli, and purified as a His6-tagged fusion in high yield. This enzyme is one of the most active XRs thus far characterized and may be used for the in vitro production of xylitol.

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