Purification and characterization of an intracellular lipase from pleopods of whiteleg shrimp (Litopenaeus vannamei)

Rivera-Pérez, Crisalejandra; Toro, María de los Angeles Navarrete del; García‐Carreño, Fernando

doi:10.1016/j.cbpb.2010.10.004

Cited by 35 publications

(16 citation statements)

References 57 publications

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“…BPL was found to be stable at 100°C after 5-min of incubation, maintains about 60% of its activity. Proteins were precipitated by acetone (v/2v) and visualized by SDS-PAGE 15% (Figure 2b), Which is a special case in the world, until now no lipase supports a temperature of 70°C and 100°C [20,21,22].…”

Section: Methodsmentioning

confidence: 99%

Biochemical characterization of a new thermostable lipase from Bacillus pumilus strain

Faouzi

Bergadi

Sayari³

et al. 2015

Turk J Bioch

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Objective: Research and characterization of new thermostable lipases from bacterial strains isolated from tannery waters in the old medina of Fez. Methods: Gene which encodes the 16S ribosomal RNA for a bacterial species was amplified via PCR and sequenced (Bacillus pumilus HF544325). The extracellular lipase from B. pumilus is purified by gel filtration (Sephacryl S-200) and cation exchange chromatography (Mono S sepharose cation exchanger). The N-terminal sequences of purified Bacillus pumilus lipase were determined by automated Edman's degradation, using an Applied Biosystems 470 A protein sequencer equipped with PTH 120A analyser. The activity of lipase was examined within the pH range of 6.0-10.0 and the effect of pH on lipase stability was determined by incubating the lipase fraction in various buffer solutions ranging from 3.0 to 10.0 for 24 h at room temperature. Results:The results showed that Bacillus pumilus is a strain that produce non-inducible lipase. This enzyme has a molecular weight of 27 kDa and presents a maximal activity at pH 8 and 45°C. The 18 N-terminal amino acid residues showed a high degree of homology with other Bacillus lipase sequences. After treatment in 100°C for 5 min, the thermostable enzyme maintains 60% of its activity, which is greater than that those founded in previous works. The enzyme retained 100% of its activity after 30 min incubation at 70°C. Conclusion: This newly isolated lipase is thermostable and it has a significant difference which was observed when the biochemical properties of the Bacillus pumilus lipase were compared to others microbial lipases. The Bacillus pumilus lipase can be considered as a good candidature for industrial and biotechnological applications. Key Words: Bacillus pumilus, lipase, purification, thermostable Conflict of Interest: The authors have no conflict of interest. ÖZETAmaç: Çalışmanın amacı Fez şehrinde bulunan tabakhane sularından izole edilen bakteri suşla-rından, sıcaklığa dayanıklı yeni bir lipaz enzimini saflaştırarak karakterize etmektir. Metod: Bakteri türleri için 16S ribosomal RNA'yı kodlayan gen PCR ile çoğaltılarak sekanslanmıştır (Bacillus pumilus HF544325). Ekstraselüler lipaz jel filtrasyonu (Sefakril S-200) ve katyon değiştirici kromatografi kullanılarak (Mono S Sefaroz katyon değiştirici) B. pumilus'den saflaştırılmıştır. Saflaştırılan Bacillus pumilus lipazının N-terminal dizisi Edman degredasyon yöntemi (Applied Biosystems 470 A protein sekanslama cihazı ile PTH120A analizör) ile saptanmıştır. Lipaz aktivitesi 6.0-10.0 pH aralığında incelenmiş, pH'nın lipaz enziminin dayanık-lılığına olan etkisi lipaz fraksiyonunu pH'sı 3.0 ile 10.0 arasında değişen çeşitli tampon çözel-tilerinde 24 saat oda sıcaklığında inkübe ederek araştırılmıştır. Bulgular: Sonuçlar Bacillus pumilus suşunun indüklenmeyen lipaz enzimini ürettiğini gös-termektedir. Lipaz enzimi 27 kDa molekül ağırlığına sahip olup, en yüksek aktiviteyi pH 8 ve 45°C'de göstermektedir. Amino ucundaki 18 amino asit diğer Bacillus lipaz dizileri ile yüksek oranda ...

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Section: Methodsmentioning

confidence: 99%

Biochemical characterization of a new thermostable lipase from Bacillus pumilus strain

Faouzi

Bergadi

Sayari³

et al. 2015

Turk J Bioch

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“…Lysosomal lipases are responsible of TAGs hydrolysis stored as lipid droplet, the major endogenous source of energy, while digestive lipases hydrolyze TAGs in food. The first digestive lipase in crustaceans was identified in the crab Carcinus mediterraneus: the enzyme neither requires colipase nor bile salts to display activity in the crab digestive gland (Cherif et al, 2007), as has been confirmed in the digestive lipase of shrimp Penaeus vannamei (Rivera-Pérez et al, 2011b), which also has an intracellular lipase. This enzyme was found in muscle, pleopods, uropods and in the digestive gland; in contrast to the digestive lipase, the intracellular lipase is a non glycosylated protein with a molecular mass of 196 kDa (Rivera-Pérez et al, 2011a).…”

Section: Introductionmentioning

confidence: 96%

“…It is a transcript of 1186 bp coding for 362 amino acid residues and a 44.8 kDa protein. It contains a residue of serine in the active site, which is contained in the consensus sequence GXSXG; the catalytic triad is His, Asp and Ser and it belongs to the clan α/β-hydrolase (CL0028) (Rivera-Pérez et al, 2011b). Although insect lipases have been extensively studied (Shukle et al, 2009;Horne et al, 2009;Yi-Peng et al, 2008), the lipid catabolism, and the regulation of the expression of the transcripts of the crustacean lipases are still unknown, making them interesting molecules to study the transcript expression to understand their role in the digestion and energy reserves mobilization.…”

Section: Introductionmentioning

confidence: 99%

Effect of fasting on digestive gland lipase transcripts expression in Penaeus vannamei

Rivera-Pérez¹,

García‐Carreño²

2011

Marine Genomics

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“…Besides, health status and lipid catabolism are two kindly significant indicators to assess the response of organisms to the change in dietary lipid under low‐salinity stress. As the key enzymes of hydrolysis of triglycerides and mitochondrial β‐oxidation of long‐chain fatty acids, triacylglycerol lipase (TGL) and carnitine palmitoyl transferase‐1 (CPT‐1) have been researched in several reports of crustacean (Chen, Li, Li et al., ; Rivera‐Pérez & García‐Carreño, ). Tumour necrosis factor‐α (TNF‐α) can regulate many cellular processes including immune function, inflammation, apoptosis, cell differentiation, proliferation and the activation of various components of the immune system (Goetz, Planas, & MacKenzie, ; Sun et al., ; Xiao et al., ).…”

Section: Introductionmentioning

confidence: 99%

Effect of dietary lipid level on growth, lipid metabolism and health status of the Pacific white shrimpLitopenaeus vannameiat two salinities

Liu

et al. 2017

Aquacult Nutr

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Three isonitrogenous diets containing 60 g kg -1, 90 g kg -1 or 120 g kg -1 lipid were formulated and fed to the Litopenaeus vannamei (2.00 ± 0.08 g) under two salinities (25 or 3 psu) in triplicate for 8 weeks. Shrimp fed 90 g kg -1 lipid had higher weight gain and specific growth rate than shrimp fed the other two diets regardless of salinity, and the hepatosomatic index increased with increasing dietary lipid at both salinities. The shrimp at 3 psu had significantly lower survival and ash content, higher condition factor, weight gain and specific growth rate than the shrimp at 25 psu. Increasing dietary lipid level induced the accumulation of serum MDA regardless of salinity, and at 3 psu, it reduced the serum GOT and GPT activities and the mRNA expression of TNF-α in intestine and gill of L. vannamei. The hepatopancreatic triacylglycerol lipase (TGL) and CPT-1 mRNA expression showed the highest value in shrimp fed 90 g kg -1 lipid diet at 3 psu. This study indicates that 120 g kg -1 dietary lipid may negatively affect the growth and induce oxidative damage in shrimp, but can improve immune defence at low salinity; 60 g kg -1 dietary lipid cannot afford the growth and either has no positive impact on the immunology for L. vannamei at 3 psu. K E Y W O R D Sgrowth, health status, histology, lipid, Litopenaeus vannamei, salinity | INTRODUCTIONThe Pacific white shrimp Litopenaeus vannamei is a popular species for commercial aquaculture production in the world because of its high tolerance of salinity from 1 to 50 practical salinity unit (psu) (Cheng, Hu, Liu, Zheng, & Qi, 2006;Pante, 1990;Saoud, Davis, & Rouse, 2003).However, poor survival and growth performance (Diaz, Farfan, Sierra, & Re, 2001;Li et al., 2007), low stress tolerance and disease resistance (Li et al., 2007Lin & Chen, 2001, 2003 Hwang, 2008). Therefore, the provision of sufficient nutrients and energy through dietary manipulation is effective to enhance the ability for L. vannamei to adapt to low salinity (Li et al., 2011Schwartz, Austen, & Wasserman, 1979;Tseng & Hwang, 2008;Welcomme & Devos, 1991).Lipids have the highest energy density among the three major nutri- In this study, two levels of salinity were set at 3 psu and 25 psu.The former is the salinity usually used for L. vannamei culture in inland saline water, and the latter is the salinity close to the isotonic point | MATERIALS AND METHODS | Experimental dietsThree isonitrogenous practical diets (380 g kg -1 crude protein) were formulated with three levels of crude lipid (60, 90 or 120 g kg).Before adding oil (fish oil: soybean oil = 1:1), all dry ingredients were finely ground and mixed thoroughly. The mixture was dissolved by adding water (250 ml/kg) and then extruded into 2-mm-diameter pellets using a meat grinder. The scattered pellets were air-dried at room temperature to <100g dry matter/kg moisture. Pellets were sieved to various sizes and stored at −20°C until use. Ingredient and proximate composition of the experimental diets are presented in Table 1. adding seawater a...

show abstract

Purification and characterization of an intracellular lipase from pleopods of whiteleg shrimp (Litopenaeus vannamei)

Cited by 35 publications

References 57 publications

Biochemical characterization of a new thermostable lipase from Bacillus pumilus strain

Biochemical characterization of a new thermostable lipase from Bacillus pumilus strain

Effect of fasting on digestive gland lipase transcripts expression in Penaeus vannamei

Effect of dietary lipid level on growth, lipid metabolism and health status of the Pacific white shrimpLitopenaeus vannameiat two salinities

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