Partial Characterization of a Lipase from Gypsy Moth (<i>Lymantria dispar</i> L.) Larval Midgut

Mrdaković, Marija; Lazarević, Jelica; Perić-Mataruga, Vesna; Ilijin, Larisa; Vlahović, Milena

doi:10.3409/fb56_1-2.103-110

Cited by 20 publications

(15 citation statements)

References 62 publications

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“…High temperature may interfere with hydrogen bond of enzyme causing denaturation of this enzyme [32]. Similar results have been shown in other insect lipases, as in gypsy moth Lymantria dispar [40], Rhynchophorus palmarum [41] and N . aenescens [16].…”

Section: Discussionsupporting

confidence: 61%

Purification and characterization of fat body lipase from the Greater Wax MothGalleria mellonella(Lepidoptera: Pyralidae)

Mahdy

Momen

El-Bar

et al. 2019

Preprint

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43Lipid mobilization and transport in insects is under investigation, especially 44 lipases and lipophorin because of their roles in energy production and transport of 45 lipids at flying activity. The present study has been conducted to purify 46 intracellular fat body lipase for the first time, from last larval instar of Galleria 47 mellonella. Purification methods by combination of ammonium sulfate 48 precipitation and gel filtration using Sephadex G-100 demonstrated that the 49 amount of protein and the specific activity of fat body lipase were 50 0.008633±0.000551 mg/ml and 1.5754±0.1042 μmol/min/mg protein, respectively, 51 with a 98.9 fold purity and recovery of 50.81%. Hence, the sephadex G-100 step 52 was more effective in purification process. SDS-PAGE and zymogram revealed 53 that fat body lipase showed two monomers with molecular weights of 178.8 and 54 62.6 kDa. Furthermore biochemical characterization of fat body lipase was carried 55 out through testing its activities against several factors such as; different 56 temperatures, pH ranges, metal ions and inhibitors ending by determination of their 57 kinetic parameters with the use of p-Nitrophenyl butyrate (PNPB) as a substrate. 58The highest activities of enzyme were determined at the temperature ranges of 35-59 37°C and 37-40°C and pH ranges of 7-9 and 7-10. The partially purified enzyme 60 showed significant stimulation by Ca 2+ , K + and Na + metal ions indicating that fat 61 body lipase is metalloproteinase. Additionally, lipase activity was strongly 4 62 inhibited by some inhibitors; phenylmethylsulfony fluoride (PMSF), ethylene-63 diaminetetractic acid (EDTA) and ethylene glycoltetraacetic acid (EGTA) 64 providing an evidence of presence of serine residue and activation of enzymes by 65 metal ions. Kinetic parameters were 301.95mM K m and 0.316 Umg -1 V max . By 66 considering the purification of fat body lipase from larvae and using some 67 inhibitors especially ion chelating agents, it is suggested to develop this study by 68 using lipase inhibitors to reach a successful control of Galleria mellonella in the 69 near future. 70 Introduction 73 Lipids are utilized efficiently by insects as substrate for reproduction 74 embryogenesis, metamorphosis and flight. The importance of lipids to insects had 75 been recognized by several authors [1,2]. 76 Lipids are large and heterogeneous group of substances which are insoluble 77 in water but soluble in nonpolar solvents. Some lipids contain fatty acids (e.g., fats 78 and waxes) while others lack them (e.g., terpenes). Lipids containing fatty acids 79 involve several groups; sphinogolipids, glycerolipids, glycerophospholipids, sterol 80 lipids, saccahrolipids, prenol lipids and polykiteds [3]. Others that contain 5 81 metabolites play an important role in biological reactions [4]. Lipids containing 82 fatty acids present as storage lipids and membrane lipids. Glycolipids and 83 phospholipids are included in membrane lipids, while the storage lipids represented 84 by fats in adipose tissue of animals,...

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Section: Discussionsupporting

confidence: 61%

Purification and characterization of fat body lipase from the Greater Wax MothGalleria mellonella(Lepidoptera: Pyralidae)

Mahdy

Momen

El-Bar

et al. 2019

Preprint

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“…After 2 h of incubation, 60% of activity of PVL was lost at pH 10. Lipase of scorpion S. maurus lost only 35% of its activity after 4 h incubation at pH 11 and pH under 5 (Zouari et al 2005), as well as lipase from the Asian gypsy moth Lymantria dispar (Mrdakovic et al 2008) and squid O. bartramii (Sukarno et al 1996). Lipase activity decreased in Rodnius prolixus and Cephaloleia presignis at pH>8.0 (Arreguín-Espinosa et al 2000; Grillo et al 2007); these evidences showed that the pH stability of lipases differs between species.…”

Section: Discussionmentioning

confidence: 99%

Purification and Biochemical Characterization of Digestive Lipase in Whiteleg Shrimp

2010

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“…Enzyme activities were measured in triplicate and controls without enzyme or without substrate were included. Lipase activity was determined using p-nitrophenyl caprylate as the substrate, by continuous monitoring of the release of p-nitrophenol at 410 nm (Mrdaković et al, 2008). Specific enzyme activities were expressed as U/mg of midgut protein, and one unit of enzyme activity is defined as the amount of enzyme that liberates 1 µmol of reaction product per minute, under the given assay conditions.…”

Section: Enzyme Assaysmentioning

confidence: 99%

Response of Lymantria dispar (Lepidoptera: Lymantriidae) larvae from differently adapted populations to allelochemical stress: Effects of tannic acid

Mrdaković¹,

Perić-Mataruga²,

Ilijin³

et al. 2013

Eur. J. Entomol.

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Abstract. The effects of tannic acid on mean values and genetic variation in fitness-related traits (mass, relative growth rate) and specific activities of digestive enzymes (total proteases, -glucosidase and lipase), and genetic variation in their plasticity, were investigated in fifth instar larvae of Lymantria dispar L. (Lepidoptera: Lymantriidae) originating from two populations with different host use histories (oak and locust-tree). The two populations did not differentiate with respect to fitness-related traits, i.e. adverse effects of tannic acid were similar in both populations. However, Robinia larvae, which originated from the locust-tree forest, were characterized by higher total protease and lipase activity and lower -glucosidase activity than Quercus larvae, which originated from the oak forest. Higher plasticity of lipase and lower plasticity of -glucosidase in response to tannic acid were also recorded. Quantitative genetic analysis revealed mostly significant expression of genetic variation in the examined traits and trait plasticity, suggesting the potential for evolution of adaptive plastic responses to new environmental conditions and presence of a stressor. The genetic correlations observed between the environments significantly differed from "one", which indicates there are no constraints on the evolution of trait plasticity.

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Partial Characterization of a Lipase from Gypsy Moth (Lymantria dispar L.) Larval Midgut

Cited by 20 publications

References 62 publications

Purification and characterization of fat body lipase from the Greater Wax MothGalleria mellonella(Lepidoptera: Pyralidae)

Purification and characterization of fat body lipase from the Greater Wax MothGalleria mellonella(Lepidoptera: Pyralidae)

Purification and Biochemical Characterization of Digestive Lipase in Whiteleg Shrimp

Response of Lymantria dispar (Lepidoptera: Lymantriidae) larvae from differently adapted populations to allelochemical stress: Effects of tannic acid

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