Lipolytic Activity by Oral Pleuropneumonia-Like (Mycoplasma) Organisms

Cole, Barry C.; Pease, Phyllis

doi:10.1099/00221287-47-2-171

Cited by 8 publications

(3 citation statements)

References 12 publications

(9 reference statements)

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“…Although the mycoplasma membrane is rich in essential enzymes, lipase activity has been detected only in the soluble fraction of disrupted cells from Mycoplasma gallisepticum, Acholeplasma laidlawii (37), and mycoplasmas isolated from human saliva (6). Esterase activity has also been detected by the histochemical staining of crude lysates of 22 Mycoplasma and Acholeplasma species (30).…”

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confidence: 99%

Mycoplasma hyopneumoniae p65 Surface Lipoprotein Is a Lipolytic Enzyme with a Preference for Shorter-Chain Fatty Acids

2004

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Mycoplasma hyopneumoniae is the most significant bacterial pathogen of the respiratory tract of swine. p65 is an immunodominant surface lipoprotein of M. hyopneumoniae that is specifically recognized during disease. Analysis of the translated amino acid sequence of the gene encoding p65 revealed similarity to the GDSL family of lipolytic enzymes. To examine the lipolytic activity of p65, the gene was cloned and expressed in Escherichia coli after truncation of the prokaryotic lipoprotein signal sequence and mutagenesis of the mycoplasma TGA tryptophan codons. After treatment with thrombin, the recombinant glutathione S-transferase (GST)-p65 protein yielded a 66-kDa fusion protein cleavage product corresponding in size to the mature p65 protein. The esterase activity of recombinant GST-p65 was indicated by the formation of a cleared zone on tributyrin agar plates and the hydrolysis of p-nitrophenyl esters of caproate (pNPC) and p-nitrophenyl esters of palmitate (pNPP). Lipase activity was indicated by the hydrolysis of the artificial triglyceride 1,2-O-dilauryl-rac-glycero-3-glutaric acid resorufin ester. Using pNPC and pNPP as substrates, recombinant GST-p65 had optimal activity between pHs 9.2 and 10.2 and at a temperature higher than 39°C. Calcium ions did not increase the activity of recombinant GST-p65. Rabbit anti-p65 antibodies inhibited the activity of recombinant GST-p65 and also inhibited the growth of M. hyopneumoniae in vitro. Examination of the kinetic parameters of recombinant GST-p65 for the hydrolysis of pNPC and pNPP indicated a preference for the shorter fatty acid chain of pNPC. The physiological and/or pathogenic role of mycoplasma lipolytic enzymes has not been determined, but they are likely to play an important role in mycoplasmas' nutritional requirements for long-chain fatty acids and may reduce the function of lung surfactants in mycoplasma-induced respiratory diseases. This is the first report of the lipolytic activity of a lipid-modified surface immunogen of a mycoplasma.Mycoplasma hyopneumoniae is the causative agent of porcine enzootic pneumonia (25). The high prevalence of this disease incurs substantial economic losses in the pig industry throughout the world. In the absence of a cell wall, proteins at the cell surface are essential for the survival and colonization of mycoplasmas within the host. A comparatively large proportion of the mycoplasma genome encodes proteins that are putatively modified by the addition of a lipid moiety, to permit attachment to the cell membrane and exposure to the extracellular milieu (5,11,17,40). These surface-exposed lipoproteins are among the primary targets of the host humoral immune response. Despite the pathogenic significance of M. hyopneumoniae, there is limited understanding of the major immunogens or their functions. Further understanding of the functional role of these proteins is likely to be essential to improving the control of mycoplasmoses.Previous work by Wise and Kim (47) identified three lipoproteins in M. hyopneumoniae strain J. T...

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Mycoplasma hyopneumoniae p65 Surface Lipoprotein Is a Lipolytic Enzyme with a Preference for Shorter-Chain Fatty Acids

2004

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“…pneumoniae, and T-mycoplasma could be demonstrated by overlaying agar cultures with a saturated ammonium sulfate solution. This is the first demonstration to substantiate the proteolytic activities by human fermentans are already known [1,2]. Repeated attempts failed to detect these activities with M. orate 1.…”

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confidence: 68%

“…BLM supplemented with 0.4% skimmed milk was used in 5 ml quantities in screw-capped test tubes of 6 ml capacity, and SMC and SMT supplemented with 1, 2, 3, and 4% skimmed milk, which were plated as mentioned above, to observe the decomposition of skimmed milk. Cole's medium [1] supplemented with egg yolk was plated as above, to observe egg yolk reaction. Three milliliters of BLM containing 20% gelatin was placed in cotton-plugged test tubes (1 x 10 cm), to observe liquefaction of gelatin.…”

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Proteolytic Activities of Human Mycoplasmas

Watanabe

Mishima

Horikawa

1973

Japanese Journal of Microbiology

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Chemical analyses, local Shwartzman reactivity, and body weight-decreasing activity of aqueous-phenol extracts of Mycoplasma salivarium cells

Totsuka

Shibata

Watanabe

1990

Antonie van Leeuwenhoek

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Aqueous-phenol extracts of Mycoplasma salivarium ATCC 23064 cells (APM) showed demonstrable differences from lipopolysaccharides (LPSs) of Veillonella rodentium ATCC 17743. These were as follows: smaller amounts of amino sugars and an absence of 2-keto-3-deoxyoctonate; local Shwartzman reactivity and body weight-decreasing activity, even though the activities were rather weak compared with those of LPSs. Therefore, phenol-water extractive components of Mycoplasma salivarium might be of pathogenic importance in mediating damaging effects on the periodontium.

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Lipolytic Activity by Oral Pleuropneumonia-Like (Mycoplasma) Organisms

Cited by 8 publications

References 12 publications

Mycoplasma hyopneumoniae p65 Surface Lipoprotein Is a Lipolytic Enzyme with a Preference for Shorter-Chain Fatty Acids

Mycoplasma hyopneumoniae p65 Surface Lipoprotein Is a Lipolytic Enzyme with a Preference for Shorter-Chain Fatty Acids

Proteolytic Activities of Human Mycoplasmas

Chemical analyses, local Shwartzman reactivity, and body weight-decreasing activity of aqueous-phenol extracts of Mycoplasma salivarium cells

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