Record Details
A physiological approach to a rapid method for identifying C. botulinum
ScholarsArchive at Oregon State University
| Field | Value |
|---|---|
| Title | A physiological approach to a rapid method for identifying C. botulinum |
| Names |
Dyer, John Kaye
(creator) Anderson, Arthur W. (advisor) |
| Date Issued | 1967-08-09 (iso8601) |
| Note | Graduation date: 1968 |
| Abstract | Disc gel electrophoresis of the vegatative cell -free extracts of strains of Clostridium botulinum types A, B, C, E, and F and the related nontoxic group showed limited value as a means for identifying these closely related microorganisms, since separation, though not consistent in all cases, could only be based on the number of protein fractions in the gel, Enzyme staining of the protein -laden polyacrylamide gels of the strains showed single or multiple molecular forms for malic (NAD and NADP), isocitric (NAD), succinic (NAD), and lactic (NAD) dehydrogenases and alkaline phosphatase. Analyzing the enzyme patterns of the strains revealed that most of these enzyme systems are useful for distinguishing the types and the nontoxic strains. A method which allowed two samples to he run in the same polyacrylamide gel showed that the differences between the total 1;c--1\ protein patterns of two strains can be demonstrated clearly. A type of iron bound protein (ferredoxin) was isolated from C. botulinum using a modification of the method recommended by L. E. Mortenson for isolating ferredoxin from Clostridium pasteurianum. The protein exhibited maximum absorption in the ultraviolet region near 260 mμ. Portions of the isolated iron bound protein were separated by disc electrophoresis, and following specific iron bound protein staining, showed a positive reaction in the same position in the gel column as first demonstrated using cell -free extract. Evidence accumulated using cell -free extract of C. botulinum suggests that pyruvate is metabolized through a phosphoroclastic system as demonstrated in other clostridia. It is probable that the ferredoxin has the important role of electron mediator between pyruvic oxidase and hydrogenas a for hydrogen evolution and acetyl phosphate formation. A purposed system for the synthesis of aspartate and glutamate in C. botulinum incorporating the above enzymes including those of the phosphoroclastic system in a partial citric acid cycle and glyoxylate bypass was described. |
| Genre | Thesis/Dissertation |
| Topic | Electrophoresis |
| Identifier | http://hdl.handle.net/1957/46576 |
