Bacterial Lipase Activity Research Articles

High bacterial colonization and lipase activity in microcomedones.

BackgroundAlthough acne vulgaris has a multifactorial aetiology, comedogenesis and bacteria colonization of the pilosebaceous unit are known to play a major role in the onset of inflammatory acne lesions. However, many aspects remain poorly understood such as where and when is the early stage of the Propionibacterium acnes colonization in follicular unit? Our research aimed at providing a precise analysis of microcomedone's structure to better understand the interplay between Propionibacterium acnes and follicular units, and therefore, the role of its interplay in the formation of acne lesions.MethodsMicrocomedones were sampled using cyanoacrylate skin surface stripping (CSSS). Their morphology was investigated with multiphoton imaging and their ultrastructure with scanning electron microscopy (SEM) and transmission electron microscopy (TEM). Bacterial lipase activity in the microcomedones was quantified using a dedicated enzymatic test as well as a Fourier Transform Infra‐Red (FTIR) analysis. The porphyrin produced by bacteria was analysed with HPTLC and fluorescence spectroscopy.ResultsThe imaging analysis showed that microcomedones' structure resembles a pouch, whose interior is mostly composed of lipids with clusters of bacteria and whose outer shell is made up of corneocyte layers. The extensive bacteria colonization is clearly visible using TEM. Even after sampling, clear lipase activity was still seen in the microcomedone. A high correlation, r = .85, was observed between porphyrin content measured with HPTLC and with fluorescence spectroscopy. These observations show that microcomedones, which are generally barely visible clinically, already contain a bacterial colonization.

Shiunko and Chuoko, topical Kampo medicines, inhibit the expression of gehA encoding the extracellular lipase in Cutibacterium acnes.

Antimicrobial agents have been used for eradication of Cutibacterium (formerly Propionibacterium) acnes that is an exacerbation factor of the skin disease acne vulgaris. However, the use of antibiotics is associated with an increased risk of promoting the emergence of resistant bacteria and leading to skin dysbiosis. Traditional Japanese Kampo medicines, such as Keigairengyoto, are used to treat acne. However, there is incomplete understanding regarding their functional mechanism in treatment of acne. In this study, we examined the antimicrobial and anti-lipase activity of the Kampo medicines used empirically for acne treatment. Three oral medicines, Keigairengyoto, Seijoboufuto and Jumihaidokuto, were found to inhibit the growth of C.acnes and decrease the lipase activity. Especially, Keigairengyoto caused remarkable decrease of bacterial lipase activity. Furthermore, topical medicines such as Shiunko and Chuoko significantly decreased the lipase activity in a dose-dependent manner, without inhibiting C.acnes growth. The topical medicines were found to inhibit the expression of gehA, which codes for extracellular lipase. Our results indicate that Shiunko and Chuoko have potential as effective acne therapeutic agents, especially because they do not promote the emergence of antimicrobial-resistant bacteria and skin dysbiosis.

Bacterial lipolysis of immune-activating ligands promotes evasion of innate defenses

Commensal and pathogenic bacteria hydrolyze host lipid substrates with secreted lipases and phospholipases for nutrient acquisition, colonization, and infection. Bacterial lipase activity on mammalian lipids and phospholipids can promote release of free fatty acids from lipid stores, detoxify antimicrobial lipids, and facilitate membrane dissolution. The gram-positive bacterium Staphylococcus aureus secretes at least two lipases, Sal1 and glycerol ester hydrolase (Geh), with specificities for short- and long-chain fatty acids, respectively, each with roles in the hydrolysis of environmental lipids. In a recent study from our group, we made the unexpected observation that Geh released by S. aureus inhibits activation of innate immune cells. Herein, we investigated the possibility that S. aureus lipases interface with the host immune system to blunt innate immune recognition of the microbe. We found that the Geh lipase, but not other S. aureus lipases, prevents activation of innate cells in culture. Mutation of geh leads to enhancement of proinflammatory cytokine production during infection, increased innate immune activity, and improved clearance of the bacterium in infected tissue. These in vitro and in vivo effects on innate immunity were not due to direct functions of the lipase on mammalian cells, but rather a result of inactivation of S. aureus lipoproteins, a major pathogen-associated molecular pattern (PAMP) of extracellular gram-positive bacteria, via ester hydrolysis. Altogether, these studies provide insight into an adaptive trait that masks microbial recognition by innate immune cells through targeted inactivation of a broadly conserved PAMP.

Enhancement of lipase activity for the oily wastewater treatment by an electrostimulation process

The efficacy of electrostimulation on bacterial lipase activity and COD removal was studied using a laboratory bioelectrochemical system.

Extracellular enzymes of cold-adapted bacteria from Arctic sea ice, Canada Basin

A total of 338 aerobic heterotrophic bacterial strains were isolated from Arctic sea ice, Canada Basin (77°30′N–80°12′N). The capability of the isolates to produce protease, lipase, amylase, chitinase, β-galactosidase, cellulase and/or agarase was investigated. Isolates that were able to degrade tributyrin, skim milk, starch, lactose and chitin accounted for 71.6, 65.7, 38.5, 31.6 and 16.9% of sea ice strains, respectively. Lipase producers and/or protease producers were phylogenetically widespread among the isolated strains. Starch and/or lactose hydrolytic strains were mainly distributed among Colwellia, Marinomonas, Pseudoalteromonas, Pseudomonas and Shewanella isolates. Pseudoalteromonas tetraodonis, Pseudoalteromonas elyakovii, Bacillus firmus and Janibacter melonis isolates all have the ability to degrade chitin. Only some strains belonging to Pseudoalteromonas genus scored positive for agarase (6) and cellulose (9). The temperature dependences for lipase activities were determined for five psychrophilic and six psychrotolerant bacteria. At low temperatures, the psychrophilic bacterial lipase activity was not significantly higher than psychrotolerant bacterial lipase, though all lipases showed remarkably high activity with 10–36% residual activity at 0°C.

Lipid biomarkers and bacterial lipase activities as indicators of organic matter and bacterial dynamics in contrasted regimes at the DYFAMED site, NW Mediterranean

This study investigated the relationships between dissolved organic matter (DOM) composition and bacterial dynamics on short time scale during spring mesotrophic (March 2003) and summer oligotrophic (June 2003) regimes, in a 0–500 m depth water column with almost no advection, at the DYFAMED site, NW Mediterranean. DOM was characterized by analyzing dissolved organic carbon (DOC), colored dissolved organic matter (CDOM) and lipid class biotracers. Bacterial dynamic was assessed through the measurement of in situ bacterial lipase activity, abundance, production and bacterial community structure. We made the assumption that by coupling the ambient concentration of hydrolysable acyl-lipids with the measurement of their in situ bacterial hydrolysis rates (i.e. the free fatty acids release rate) would provide new insights about bacterial response to change in environmental conditions. The seasonal transition from spring to summer was accompanied by a significant accumulation of excess DOC (+5 μM) (ANOVA, p <0.05, n =8) in the upper layer (0–50 m). In this layer, the free fatty acids release rate to the bacterial carbon demand (BCD) ratio increased from 0.6±0.3 in March to 1.3±1.0 in June (ANOVA, p <0.05, n =8) showing that more uncoupling between the hydrolysis of the acyl-lipids and the BCD occurred during the evolution of the season, and that free fatty acids contributed to the excess DOC. The increase of lipolysis index and CDOM absorbance (from 0.24±0.17 to 0.39±0.13 and from 0.076±0.039 to 0.144±0.068; ANOVA, p <0.05, n= 8, respectively), and the higher contribution of triglycerides, wax esters and phospholipids (from <5% to 12–31%) to the lipid pool reflected the change in the DOM quality. In addition to a strong increase of bacterial lipase activity per cell (51.4±29.4–418.3±290.6 Ag C cell −1 h −1 ), a significant percentage of ribotypes (39%) was different between spring and summer in the deep chlorophyll maximum (DCM) layer in particular, suggesting a shift in the bacterial community structure due to the different trophic conditions. At both seasons, in the chlorophyll layers, diel variations of DOM and bacterial parameters reflected a better bioavailability and/or DOM utilization by bacteria at night (the ratio of free fatty acids release rate to bacterial carbon demand decreased), most likely related to the zooplankton trophic behaviour. In mesotrophic conditions, such day/night pattern was driving changes in the bacterial community structure. In more oligotrophic period, diel variations in bacterial community structure were depth dependent in relation to the strong summer stratification.

083 Effect of nadifloxacin on poly-morphonuclear leukocyte functions and bacterial lipase activity

083 Effect of nadifloxacin on poly-morphonuclear leukocyte functions and bacterial lipase activity

Survival of Lipase during Manufacture of Nonfat Dry Milk

Extracellular lipases from several strains of Pseudomonas fluorescens were active against milk fat. The activity of P. fluorescens strain B52 lipase in reconstituted NDM using β-naphthyl caprylate as substrate was 70% of that found with skim milk; however, hydration of the NDM at 4°C for 12h resulted in 85% recovery of activity. As little as .0025% (vol/vol) of P. fluorescens spent media was detected in reconstituted skim milk powder using β-naphthyl caprylate. The process of freeze-drying or spray-drying without heating had little effect on either native or P. fluorescens B52 lipase. The bacterial lipase activity was reduced by 12 to 21% and 55 to 59% in NDM treated with low heat (72°C/16s) and high heat (110°C/2min), respectively, while the native lipases were completely inhibited by these treatments. Both native and bacterial enzymes were stable when stored in NDM for extended periods at 20°C. In a limited study of commercial NDM samples, microbial lipase was not detected, suggesting that the quality of the raw milk was sufficiently high to restrict the presence of heat-stable lipolytic enzymes.

Lipolytic Activity of Human Cutaneous Bacteria

SUMMARY: Forty-two strains of Corynebacterium acnes were compared with 20 strains of staphylococci and 10 of lipophilic diphtheroids for their lipolytic action on 14 substrates and for Tween 80 hydrolysis. All strains were isolated from normal human skin. One lipophilic diphtheroid and 33 C. acnes strains hydrolysed tributyrin. Forty-nine bacillary strains hydrolysed Tween 80. None of the other substrates was degraded. C. xerosis atcc 373 and Propioni-bacterium acnes atcc 11828 acted on tributyrin. P. acnes also hydrolysed Tween 80. The staphylococci did not act on Tween 80 but attacked tributyrin, triolein, trilinolenin, trilinolein and oil emulsions. Some cocci acted on tricaprylin, tricaprin and trilaurin; no lipolysis by this group was observed of trimyristin, tripalmitin or tristearin. Pancreatic lipase, tested by the penicylinder diffusion method for comparison, was active on the same substrates, as were the cocci. The addition of glucose, lactate and urea to media did not inhibit lipolysis by the cocci. Sodium oleate inhibited pancreatic lipase action on triolein and cottonseed oil but not on the hydrolysis of tributyrin. Coccal lipase action was not inhibited by oleate. All 74 test strains were uniformly inhibited by neomycin 5–10 μg./ml. Gram-positive cocci rather than C. acnes appear primarily responsible for bacterial lipase activity on human skin.