Directed evolution of a beta-galactosidase from Pyrococcus woesei resulting in increased thermostable beta-glucuronidase activity

2007 ◽  
Vol 77 (3) ◽  
pp. 569-578 ◽  
Author(s):  
Ai-Sheng Xiong ◽  
Ri-He Peng ◽  
Jing Zhuang ◽  
Xian Li ◽  
Yong Xue ◽  
...  
Keyword(s):  
Directed Evolution ◽  
Pyrococcus Woesei ◽  
Glucuronidase Activity ◽  
Beta Galactosidase

scholarly journals Effect of potassium deficiency on mouse kidney lysosomal enzymes

1978 ◽  
Vol 170 (2) ◽  
pp. 249-256 ◽  
Author(s):  
C E Cleveland ◽  
R T Swank
Keyword(s):  
Lysosomal Enzymes ◽  
Fold Increase ◽  
Inbred Strains ◽  
Deficient Diet ◽  
Proximal Tubule Cells ◽  
Glucuronidase Activity ◽  
Infiltrating Cells ◽  
Enzyme Molecules ◽  
Beta Galactosidase ◽  
Deficient Mice

Mice of inbred strains A/J, C57BL/6J and C57BL/6J beige were kept on a K+-deficient diet for up to 40 days to determine the magnitude and mechanism of changes in tissue lysosomal enzymes. From days 10 to 40 glucuronidase activity increased 3-fold in kidney of K+-deficient mice, but there was little effect on beta-galactosidase or acid phosphatase activity. Similar increases in kidney glucuronidase activity occurred in inbred strains known to have genetically altered control of the synthesis (A/J) and secretion (C57BL/6J beige) of glucuronidase in kidney proximal-tubule cells. Deprivation of K+ did not affect glucuronidase activity in liver, spleen, lung and brain, but there was a 2-3-FOld increase in glucuronidase activity in heart in the C57BL/6J and C57BL/6J beige strains. As shown by specific antibody titration, increased glucuronidase activity in kidney of K+-deficient mice was accompanied by accumulation of enzyme molecules. Likewise in kidney of deficient mice there was an increased rate of synthesis of glucuronidase as measured by incorporation of labelled leucine into immunoprecipitable glucuronidase. In kidney of K+-deficient mice the elevated glucuronidase activity was found in both collecting-tubule and interstitial cells of the medulla. It is probable therefore that a significant fraction of the increased kidney lysosomal synthesis and enzyme activity is due to infiltrating cells.

scholarly journals Directed Evolution of Beta-galactosidase from Escherichia coli into Beta-glucuronidase

BMB Reports ◽  
2007 ◽  
Vol 40 (3) ◽  
pp. 419-425 ◽  
Author(s):  
Ai-Sheng Xiong ◽  
Ri-He Peng ◽  
Jing Zhuang ◽  
Jin-Ge Liu ◽  
Fang Xu ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Directed Evolution ◽  
Beta Galactosidase

scholarly journals Binding of human liver hydrolases by immobilized lectins

1979 ◽  
Vol 177 (1) ◽  
pp. 175-180 ◽  
Author(s):  
M B Fiddler ◽  
Y Ben-Yoseph ◽  
H L Nadler
Keyword(s):  
Human Liver ◽  
Similar Data ◽  
Galactosidase Activity ◽  
Purification And Characterization ◽  
Ulex Europaeus ◽  
Glucuronidase Activity ◽  
Beta Galactosidase ◽  
Carbohydrate Specificities ◽  
Ulex Europaeus Lectin I

The binding of 22 human liver hydrolase activities by immobilized lectins of six different carbohydrate specificities, namely alpha-D-mannose (glucose), D-N-acetylglucosamine, D-N-acetylgalactosamine, L-fucose, alpha-D-galactose and beta-D-galactose, were examined. Differences in binding among these enzymes and within specific enzymes were observed. For example, the neutral forms of alpha-mannosidase and beta-xylosidase were bound by the Ulex europaeus lectin I (specific for L-fucose), whereas the acidic forms were not. Bandierea simplicifolia lectin (specific for alpha-galactose) bound 65% of beta-glucuronidase activity; recycling experiments demonstrated complete binding of the enzyme that had been eluted with the competitor D-galactose and no binding of the fraction that was not initially bound. These results suggested the presence of two forms of this enzyme. Similar data were obtained for acidic beta-galactosidase activity. These experiments may provide the basis for the expanded use of immobilized lectins for purification and characterization of hydrolases and other glycoproteins.

Visualization of von Willebrand Factor Multimers by Enzyme-Conjugated Secondary Antibodies

1986 ◽  
Vol 55 (02) ◽  
pp. 276-278 ◽  
Author(s):  
F Brosstad ◽  
Inge Kjønniksen ◽  
B Rønning ◽  
H Stormorken
Keyword(s):  
Von Willebrand Factor ◽  
Chromogenic Substrate ◽  
Agarose Electrophoresis ◽  
Secondary Antibodies ◽  
Von Willebrand ◽  
Willebrand Factor ◽  
Beta Galactosidase

SummaryA method for visualization of the multimeric forms of von Willebrand Factor (vWF) in plasma and platelets is described. The method is based upon: 1) Separation of the vWF multimers by SDS-agarose electrophoresis, 2) Subsequent blotting of the vWF multimers onto nitrocellulose, 3) Immunolocalization and visualization of the vWF pattern by the sequential incubation of the blot with a) primary vWF antiserum, b) peroxidase- or beta-galactosidase-conjugated secondary antibodies and a relevant chromogenic substrate.

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