Lipolysis of natural long chain and synthetic medium chain galactolipids by pancreatic lipase-related protein 2

Amara, Sawsan; Barouh, Nathalie; Lecomte, Jérôme; Lafont, Dominique; Robert, S.; Villeneuve, Pierre; De, Alain; Carrière, Frédéric

doi:10.1016/j.bbalip.2010.01.003

Cited by 43 publications

(37 citation statements)

References 35 publications

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“…The specific activity of F. solani cutinase on C8‐MGDG (984 ± 62 U/mg) represented 55% and 18% of rHPLRP2 (1786 ± 100 U/mg) and rGPLRP2 (5420 ± 85 U/mg) specific activities, respectively, on the same substrate. The microbial lipase with the highest galactolipase activity thus displayed a lower activity than the most active galactolipases identified so far 11. The galactolipase activities of cutinase and TLL were closer to those measured with HCEH and BCEH origins (Table 1).…”

Section: Resultssupporting

confidence: 60%

“…Dioctanoyl galactolipid substrates, C8‐MGDG (3‐ O ‐β‐ D ‐galactopyranosyl‐1,2‐di‐ O ‐octanoyl‐ sn ‐glycerol) and C8‐DGDG (3‐ O ‐(6‐ O ‐α‐ D ‐galactopyranosyl‐β‐ D ‐galactopyranosyl)‐1,2‐di‐ O ‐octanoyl‐ sn ‐glycerol), were synthesized as previously described 10, 11. The galactolipase activity was measured potentiometrically at 37°C and at a constant pH value of 8.0 by continuously measuring the release of free fatty acids from mechanically stirred dispersion of galactolipids, using 0.1 N NaOH as titrant and a pH‐stat apparatus (718 STAT Titrino, Metrohm).…”

Section: Methodsmentioning

confidence: 99%

“…The current biochemical knowledge on galactolipases remains however limited while lipases and phospholipases have been well characterized and are currently used in many biotechnological applications 5, 6. This is partly due to the fact that galactolipid substrates are not commercially available at large scale and low cost, and only a few laboratories have been involved in the development of specific assays for measuring galactolipase activities 7–12. Second, although the existence of galactolipid acyl hydrolases in the plant kingdom has been known for a long time, their levels are usually low and their purification from natural sources is hard to achieve.…”

Section: Introductionmentioning

confidence: 99%

“…In HPLRP2, there is a full‐length lid but its unusual conformation compared to that observed in HPL allows a larger access to the active site 23. Besides these structural features, kinetics of galactolipid hydrolysis were obtained thanks to the synthesis of medium chain MGDG and DGDG substrates and their use for setting‐up sensitive and continuous galactolipase assays 10–12.…”

Section: Introductionmentioning

confidence: 99%

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The galactolipase activity of some microbial lipases and pancreatic enzymes

Amara

Lafont

Parsiegla

et al. 2013

Euro J Lipid Sci & Tech

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Several well known microbial lipases were screened for their ability to hydrolyze synthetic medium chain monogalactosyldiacylglycerol (MGDG) and digalactosyldiacylglycerol (DGDG). Fusarium solani cutinase and Thermomyces lanuginosus lipase (TLL) were found to hydrolyze MGDG at high rates (984 AE 62 and 450 AE 41 U/mg, respectively). These activities remained however lower than those measured with pancreatic lipase-related protein 2 (PLRP2) on the same substrate. As previously observed with PLRP2, galactolipid-bile salt mixed micelles were found to be the best substrate form for microbial enzymes. The galactolipid to bile salt molar ratios for measuring maximum galactolipase activities were found to be similar to those previously established with PLRP2, suggesting that bile salts have mainly an effect on the substrate and not on the enzyme itself. The galactolipase activity of cutinase and TLL, as well as human and guinea pig PLRP2s were also measured using galactolipid monomolecular films. Enzymes having a lid (TLL and human PLRP2) were found to act at higher surface pressures than those with no lid (cutinase and guinea pig PLRP2). In silico docking of medium chain MGDG and DGDG in the active site of guinea pig PLRP2 and TLL reveals some structural analogies between these enzymes.Practical applications: Galactolipase activity already finds applications in baking process with the partial lipolysis of galactolipids present in wheat flour and the in situ production of emulsifier molecules. More generally, these enzymes could be used for the enzymatic modification of galactolipids, the main plant lipids. These lipids are considered to be the most abundant at the surface of the earth and thus constitute the largest resource of fatty acids. Fax: þ33 491 715 857Abbreviations: BCEH, bovine pancreatic carboxylester hydrolase; CEH, pancreatic carboxylester hydrolase; DGDG, digalactosyldiacylglycerol; GPLRP2, guinea pig pancreatic lipase-related protein 2; HCEH, human pancreatic carboxylester hydrolase; HPL, human pancreatic lipase; HPLRP2, human pancreatic lipase-related protein 2; MGDG, monogalactosyldiacylglycerol; NaTDC, sodium taurodeoxycholate; PLRP2, pancreatic lipase-related protein 2; TAG, triacylglycerol; TC4, tributyrin; TLL, Thermomyces lanuginosus lipase; YLLIP2, Yarrowia lipolytica lipase 442Eur.

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Section: Resultssupporting

confidence: 60%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

The galactolipase activity of some microbial lipases and pancreatic enzymes

Amara

Lafont

Parsiegla

et al. 2013

Euro J Lipid Sci & Tech

Self Cite

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“…The lipid extracts corresponding to about 25 nmol of total fatty acids were dissolved in 100 µl chloroform/methanol [2/1, (v/v)] containing 125 pmol of each internal standard. Internal standards were obtained from Avanti Polar Lipids Inc. for PG 18:0–18:0 or synthesized by D. Lafont (Amara et al ., ) for MGDG 18:0–18:0 and DGDG 16:0–16:0 or extracted from spinach thylakoid (Demé et al ., ) and hydrogenated as previously described for SQDG 16:0–18:0 (Buseman et al ., ). Lipids were then separated by HPLC and quantified by MS/MS.…”

Section: Methodsmentioning

confidence: 97%

Thermoacclimation and genome adaptation of the membrane lipidome in marine Synechococcus

Pittera

Jouhet

Breton

et al. 2017

Environmental Microbiology

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The marine cyanobacteria of the genus Synechococcus are important primary producers, displaying a wide latitudinal distribution that is underpinned by diversification into temperature ecotypes. The physiological basis underlying these ecotypes is poorly known. In many organisms, regulation of membrane fluidity is crucial for acclimating to variations in temperature. Here, we reveal the detailed composition of the membrane lipidome of the model strain Synechococcus sp. WH7803 and its response to temperature variation. Unlike freshwater strains, membranes are almost devoid of C18, mainly containing C14 and C16 chains with no more than two unsaturations. In response to cold, we observed a rarely observed process of acyl chain shortening that likely induces membrane thinning, along with specific desaturation activities. Both of these mechanisms likely regulate membrane fluidity, facilitating the maintenance of efficient photosynthetic activity. A comprehensive examination of 53 Synechococcus genomes revealed clade-specific gene sets regulating membrane lipids. In particular, the genes encoding desaturase enzymes, which is a key to the temperature stress response, appeared to be temperature ecotype-specific, with some of them originating from lateral transfers. Our study suggests that regulation of membrane fluidity has been among the important adaptation processes for the colonization of different thermal niches by marine Synechococcus.

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