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Adenosine triphosphate content of Mycobacterium leprae: effect of purification procedures
Abstract:
Various procedures to decontaminate and purify M leprae free of host tissue material resulted in total retention of their intracellular ATP and also infectiousness. The ATP content of one million M. leprae cells, isolated from either livers, spleens, or lymph nodes of infected armadillos, or a nude mouse foot pad or a human biopsy specimen, was in the range of 1.17 to 1.40 picograms. Suspensions could be decontaminated with 4% NaOH and all non-bacterial ATP could be eliminated by the combined action of trypsin, chymotrypsin, and collagenase initially, followed by Triton X-100 plus ATPase. These findings further assure that M. leprae are different from M. lepraemurium in that they can withstand even the severest purification procedures that are necessary in order for them to be used for sophisticated biochemical and metabolic studies.
Insights
Purification methods for Mycobacterium leprae (M. leprae) retain intracellular ATP and infectiousness. This confirms M. leprae
Area of Science:
- Microbiology
- Biochemistry
- Infectious Diseases
Background:
- Mycobacterium leprae (M. leprae) is the causative agent of leprosy.
- Isolating pure M. leprae for research is challenging due to host tissue contamination.
- Understanding M. leprae's biochemical properties requires pure, viable bacterial samples.
Purpose of the Study:
- To develop and validate decontamination and purification procedures for M. leprae.
- To assess the impact of purification on M. leprae's intracellular ATP content and infectiousness.
- To differentiate M. leprae from M. lepraemurium based on purification tolerance.
Main Methods:
- M. leprae were isolated from infected armadillo livers, spleens, lymph nodes, nude mouse foot pads, and human biopsy specimens.
- Purification involved decontamination with 4% NaOH.
- Non-bacterial ATP was eliminated using enzymatic digestion (trypsin, chymotrypsin, collagenase) followed by Triton X-100 and ATPase treatment.
Main Results:
- Purification procedures resulted in the complete retention of intracellular ATP in M. leprae.
- The ATP content of one million purified M. leprae cells ranged from 1.17 to 1.40 picograms.
- Purified M. leprae retained their infectiousness.
Conclusions:
- The developed purification methods effectively yield pure, viable M. leprae.
- M. leprae's ability to withstand harsh purification confirms its distinctness from M. lepraemurium.
- These findings support the use of purified M. leprae in advanced biochemical and metabolic studies.