Post by skyship on Dec 3, 2009 12:02:05 GMT -5
Here we have the catalyst enzyme from C. perfringens
that changes human blood types A, B, and AB to O UNIVERSAL
blood.
A novel E. coli expression system could provide sufficient quantities of a Clostridium perfringens enzyme capable of transforming type A[subscript]2 blood cells to type O.
In a study conducted by scientists in the United States, C. perfringens alpha-N-acetylgalactosaminidase (alphaNAG) "hydrolyzed the terminal N-acetyl-alpha-D-galactosamine from the blood type A[subscript]2 antigen producing H antigen, blood type O.
www.encyclopedia.com/doc/1G1-115763472.html
====================
Purification and characterization of a recombinant alpha-N-acetylgalactosaminidase from Clostridium perfringens.
Hsieh HY, Calcutt MJ, Chapman LF, Mitra M, Smith DS.
Department of Pathology and Anatomical Sciences, University of Missouri-Columbia, Missouri, USA.
Clostridium perfringens alpha-N-acetylgalactosaminidase (alphaNAG) hydrolyzed the terminal N-acetyl-alpha-d-galactosamine from the blood type A(2) antigen producing H antigen, blood type O. Blood type O is universally compatible in the ABO system. Purification of the native enzyme is difficult with very low yields. To obtain the enzyme in satisfactory yield, the gene encoding the clostridial enzyme was cloned in an Escherichia coli T7 expression system. A highly purified preparation of recombinant alphaNAG was obtained from cell lysates by ion-exchange chromatography and high-pressure liquid chromatography. The final preparation was homogeneous by SDS-PAGE with a molecular mass of 71.96kDa and the native molecular weight of 72.42kDa. The enzyme was highly selective for terminal N-acetylgalactosamine residues. No other significant exoglycosidase activities, particularly neuraminidase, were detected. The pH optimum of the enzyme was between 6.5 and 7.0 and activity was relatively unaffected by ionic strength. ELISA experiments demonstrated activity against blood type A(2) epitope. These characteristics were similar to those of native alphaNAG from C. perfringens. With adequate expression in E. coli, sufficient recombinant alphaNAG enzyme mass can be obtained for potential use in enzymatic conversion of human blood type A(2) red blood cells to universally transfusable type O red blood cells.
www.ncbi.nlm.nih.gov/pubmed/14965778
===================
hot dang!
===================
Enzymes That Destroy Blood Group Specificity
V. THE OLIGOSACCHARASE OF CLOSTRIDIUM PERFRINGENS
Abstract
Evidence is presented for the presence of an enzyme, α-N-acetylgalactosaminyl oligosaccharase in filtrates of Clostridium perfringens. Action of this enzyme on hog submaxillary glycoprotein results in the release of a disaccharide, characterized as 3-O-β-d-galactopyranosyl-2-acetamido-2-deoxy-d-galactose. The possibility that this oligosaccharide has resulted from transgalactosylation has been eliminated. Enzymatic treatment of several glycoproteins and blood group substances from miscellaneous sources also resulted in the production of this oligosaccharide. Since in the blood group substances N-acetylgalactosamine at the reducing end of the oligosaccharide chain is α-linked to the hydroxy amino acids of the protein core, we have here a very useful α-N-acetyl-d-galactosaminyl oligosaccharase for structural studies of the carbohydrate-protein linkage in the blood group substances and in glycoproteins.
www.jbc.org/content/247/21/6737.abstract
recombinant alphaNAG enzyme mass
So, these are expressed through the recombinant
e-coli by way of an enzyme.
Now we are getting closer to the erythrocyte
form itself.
C. Perfringen Clostridium perfringen? IS TOXIC and causes
colstridium!
=========================
BBB - Clostridium perfringens
Bad Bug Book:
Foodborne Pathogenic Microorganisms and Natural Toxins Handbook
Clostridium perfringens
......"Perfringens food poisoning is the term used to describe the common foodborne illness caused by C. perfringens. A more serious but rare illness is also caused by ingesting food contaminated with Type C strains. The latter illness is known as enteritis necroticans or pig-bel disease."......
tinyurl.com/y9bjrfa
www.fda.gov/Food/FoodSafety/Foodborneillness/
FoodborneillnessFoodbornePathogensNaturalToxins/
BadBugBook/ucm070483.htm
skyship
that changes human blood types A, B, and AB to O UNIVERSAL
blood.
A novel E. coli expression system could provide sufficient quantities of a Clostridium perfringens enzyme capable of transforming type A[subscript]2 blood cells to type O.
In a study conducted by scientists in the United States, C. perfringens alpha-N-acetylgalactosaminidase (alphaNAG) "hydrolyzed the terminal N-acetyl-alpha-D-galactosamine from the blood type A[subscript]2 antigen producing H antigen, blood type O.
www.encyclopedia.com/doc/1G1-115763472.html
====================
Purification and characterization of a recombinant alpha-N-acetylgalactosaminidase from Clostridium perfringens.
Hsieh HY, Calcutt MJ, Chapman LF, Mitra M, Smith DS.
Department of Pathology and Anatomical Sciences, University of Missouri-Columbia, Missouri, USA.
Clostridium perfringens alpha-N-acetylgalactosaminidase (alphaNAG) hydrolyzed the terminal N-acetyl-alpha-d-galactosamine from the blood type A(2) antigen producing H antigen, blood type O. Blood type O is universally compatible in the ABO system. Purification of the native enzyme is difficult with very low yields. To obtain the enzyme in satisfactory yield, the gene encoding the clostridial enzyme was cloned in an Escherichia coli T7 expression system. A highly purified preparation of recombinant alphaNAG was obtained from cell lysates by ion-exchange chromatography and high-pressure liquid chromatography. The final preparation was homogeneous by SDS-PAGE with a molecular mass of 71.96kDa and the native molecular weight of 72.42kDa. The enzyme was highly selective for terminal N-acetylgalactosamine residues. No other significant exoglycosidase activities, particularly neuraminidase, were detected. The pH optimum of the enzyme was between 6.5 and 7.0 and activity was relatively unaffected by ionic strength. ELISA experiments demonstrated activity against blood type A(2) epitope. These characteristics were similar to those of native alphaNAG from C. perfringens. With adequate expression in E. coli, sufficient recombinant alphaNAG enzyme mass can be obtained for potential use in enzymatic conversion of human blood type A(2) red blood cells to universally transfusable type O red blood cells.
www.ncbi.nlm.nih.gov/pubmed/14965778
===================
hot dang!
===================
Enzymes That Destroy Blood Group Specificity
V. THE OLIGOSACCHARASE OF CLOSTRIDIUM PERFRINGENS
Abstract
Evidence is presented for the presence of an enzyme, α-N-acetylgalactosaminyl oligosaccharase in filtrates of Clostridium perfringens. Action of this enzyme on hog submaxillary glycoprotein results in the release of a disaccharide, characterized as 3-O-β-d-galactopyranosyl-2-acetamido-2-deoxy-d-galactose. The possibility that this oligosaccharide has resulted from transgalactosylation has been eliminated. Enzymatic treatment of several glycoproteins and blood group substances from miscellaneous sources also resulted in the production of this oligosaccharide. Since in the blood group substances N-acetylgalactosamine at the reducing end of the oligosaccharide chain is α-linked to the hydroxy amino acids of the protein core, we have here a very useful α-N-acetyl-d-galactosaminyl oligosaccharase for structural studies of the carbohydrate-protein linkage in the blood group substances and in glycoproteins.
www.jbc.org/content/247/21/6737.abstract
recombinant alphaNAG enzyme mass
So, these are expressed through the recombinant
e-coli by way of an enzyme.
Now we are getting closer to the erythrocyte
form itself.
C. Perfringen Clostridium perfringen? IS TOXIC and causes
colstridium!
=========================
BBB - Clostridium perfringens
Bad Bug Book:
Foodborne Pathogenic Microorganisms and Natural Toxins Handbook
Clostridium perfringens
......"Perfringens food poisoning is the term used to describe the common foodborne illness caused by C. perfringens. A more serious but rare illness is also caused by ingesting food contaminated with Type C strains. The latter illness is known as enteritis necroticans or pig-bel disease."......
tinyurl.com/y9bjrfa
www.fda.gov/Food/FoodSafety/Foodborneillness/
FoodborneillnessFoodbornePathogensNaturalToxins/
BadBugBook/ucm070483.htm
skyship