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Effect of actinophage infection on glucose metabolism in Streptomyces griseus

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Title Effect of actinophage infection on glucose metabolism in Streptomyces griseus
Names Nishikawa, Sumie Suzuki (creator)
Pilcher, K. S. (advisor)
Date Issued 1968-04-19 (iso8601)
Note Graduation date: 1968
Abstract Glucose metabolism of phage-infected Streptomyces griseus
was examined by using the specific radioactivities of the amino
acids from the extracted protein of the host and phage as an index
of the catabolic pathways of glucose metabolism. Conventional
methods for the study of phage-infected bacteria did not suit the
host-actinophage system due to the filamentous nature of the host
and the poor adsorption properties of the phage. The present
method measured the specific activities of selected amino acids
arising from glucose-1-¹⁴C and compared them to those of the
amino acids synthesized from glucose-6-¹⁴C. A C6/C1 (ratio of
the specific activity of the amino acid arising from glucose-6-¹⁴C
to that arising from glucose-1-¹⁴C) specific activity ratio of 1.0
reflected the primary catabolism of glucose via the glycolytic
pathway. A considerably higher ratio was expected if the pentose
phosphate pathway constituted the major catabolic route since carbon
one of glucose is lost at an early stage in the latter pathway. The
specific activities were obtained on protein hydrolysates by automatic
chromatographic procedures using the Beckman/Spinco Model 120B
amino acid analyzer connected to a Packard Series 320E Flow Monitor.
The procedure was tested on two cultures, Saccharomyces
cerevisiae and Pseudomonas stutzeri whose widely divergent
catabolic pathways had been already established elsewhere. The
former dissimilates glucose primarily by glycolysis while the latter
exhibits strong Entner-Doudoroff and pentose phosphate pathway
activity. Using glutamic acid as a reference amino acid, a C6/C1
ratio of 1.13 and 16.3 were obtained for S. cerevisiae and P.
stutzeri, respectively. These results indicated that the ratios could
be used as a measure of the route of carbohydrate metabolism.
Prior to determining the specific activity ratio for uninfected
S. griseus, the route of glucose catabolism employed by this organism
was ascertained by high-resolution radiorespirometry and the results
indicated a predominant glycolytic activity. The pattern observed
was similar to that found nine years earlier. The predominating
glycolysis corresponded well to the C6/C1 ratio of 1.23
obtained for S. griseus using glutamic acid as the reference amino
acid. To compare the route involved in the glucose catabolism of
the S. griseus phage-infected system, the specific activity ratios
for the amino acids from purified actinophage protein were determined.
A complete amino acid composition (with the exception of
tryptophan) was determined for this actinophage; however, for
specific activity ratio determinations, only the acidic amino acids
were analyzed. Again using glutamic acid for reference a ratio of
1.74 for the phage was obtained. The higher ratios obtained for
phage protein as compared with host protein indicated an increased
loss of C-1 after phage infection and during biosynthesis. The increased
loss of C-1 in the phage infected system may have reflected
a change in the metabolic patterns of the host to meet the
demands for materials necessary for phage biosynthesis, primarily
pentose for viral DNA. The direction of the switch in glucose
dissimilation differs from that reported in the phage-infected
Escherichia coli system and may be attributed to differences in
the mechanisms by which the two systems meet the demand for
pentose synthesis.
Genre Thesis/Dissertation
Topic Bacteriophages
Identifier http://hdl.handle.net/1957/46427

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