NOPR Community:

Purification and characterization of structural and functional properties of two lectins from a marine sponge Spheciospongia vesparia

2013-12-01T00:00:00Z

Title: Purification and characterization of structural and functional properties of two lectins from a marine sponge Spheciospongia vesparia Authors: Fenton, Bertha; Espinosa, Roberto Arreguín; Contreras, Edgar Vázquez; Lozano, Barbarin Arreguín; Sánchez, Norma Sánchez; Hernández, Enrique García; Galindo, Edgar Zenteno Abstract: The purification, structural and functional characterization of two different lectins (named Svl-1 and Svl-2) has been reported from the marine sponge Spheciospongia vesparia. Purification procedure includes ammonium sulfate precipitation, combined with chromatography including Octyl-Sepharose-(NH₄)SO₄ hydrophobic column and DEAE-Toyopearl anion-exchange column using a high performance liquid chromatography. The similarities in function, specificity for saccharides, molecular weight, amino acid content and the N-terminal sequence of two lectins suggest that these proteins are isolectins. Amino acid composition and fluorescence analyses reveal that they contain an intrachain disulfide bridge, which might contribute to their high thermal stability. Furthermore, the purified lectins exhibit antibacterial activity against the gram-negative bacteria Pseudomonas aeruginosa and E. coli, indicating that they may be involved in a recognition strategy and may play a role in the defense response function of the sponge. This is the first report on the isolation of lectins from the S. vesparia. The purified lectins represent a potential possible candidate for future application in the recognition or treatment of cancer cells. Page(s): 562-569

Puriﬁcation and characterization of 6-phosphogluconate dehydrogenase (6-PGD) from grass carp (Ctenopharyngodon idella) hepatopancreas

2013-12-01T00:00:00Z

Title: Puriﬁcation and characterization of 6-phosphogluconate dehydrogenase (6-PGD) from grass carp (Ctenopharyngodon idella) hepatopancreas Authors: Sun, Lin-Dan; Luo, Zhi; Hu, Wei; Zhuo, Mei-Qin; Zheng, Jia-Lang; Chen, Qi-Liang; Liang, Xu-Fang; Xiong, Bang-Xi Abstract: 6-Phosphogluconate dehydrogenase (6-PGD, E.C.: 1.1.1.44) was purified and characterized from the hepatopancreas of grass carp (Ctenopharyngodon idella) for the first time. Grass carp represents the second largest aquaculture industry in the world after silver carp, constituting 14.7% of the world aquaculture production, with an average annual increase of 14% in China, mainly as a source of food. The purification procedure involved a single 2’, 5’-ADP-Sepharose 4B affinity chromatographic step by using different elution buffers. The enzyme was purified 309-fold with a specific activity of 5.259 U/mg protein and yield of 68%. The purity and subunit molecular weights of the 6-PGD were checked on SDS-PAGE and purified enzyme showed a single band on the gel. The subunit molecular mass was 57 kDa, with an optimum pH, temperature and ionic strength at 7.96, 50^oC and 100 mM Tris-HCl, respectively. The K_m values of 6-PGA and NADP⁺were 0.019 and 0.0052 mM, respectively, while V_m of 6-PGA and NADP⁺ was 0.69 U/ml. Dissociation constants (K_i) for 6-PGA and NADP⁺were 2.05 and 0.12 mM, respectively. NADPH inhibited the enzyme in a competitive manner and its K_i value was 0.032 mM. The Cu²⁺, Zn²⁺, Cd²⁺ and Al³⁺ showed inhibitory effects on the enzyme with IC₅₀ values of 0.293, 0.099, 0.045 and 1.526 mM, respectively. All tested metals inhibited the enzyme in a competitive manner, indicating that these metals might be toxic even at low concentrations for the 6-PGD. As the fish is one of valuable foodstuff of animal sources for human consumption, under certain environmental conditions, metal ions accumulated in fish up to a lethal concentration may be harmful for human health. Therefore, it is impending to reduce the concentration of metal ions in contaminated lakes and rivers for fishery and also for human health. Page(s): 554-561

Carotenoid genes transcriptional regulation for astaxanthin accumulation in fresh water unicellular alga Haematococcus pluvialis by gibberellin A3 (GA₃)

2013-12-01T00:00:00Z

Title: Carotenoid genes transcriptional regulation for astaxanthin accumulation in fresh water unicellular alga Haematococcus pluvialis by gibberellin A3 (GA₃) Authors: Gao, Zhengquan; Meng, Chunxiao; Gao, Hongzheng; Li, Yan; Zhang, Xiaowen; Xu, Dong; Zhou, Shitan; Liu, Banghui; Su, Yuanfeng; Ye, Naihao Abstract: The fresh water unicellular alga Haematococcus pluvialis is a promising natural source of astaxanthin. The present study investigated the transcriptional expression of carotenoid genes for astaxanthin accumulation in H. pluvialis using real-time fluorescence quantitative PCR (qRT-PCR). With treatments of 20 and 40 mg/L of gibberllin A₃ (GA₃), five genes ipi-1, ipi-2, psy, pds and bkt2 were up-regulated with different expression profiles. GA20 (20 mg/L of GA₃) treatment had a greater effect on transcriptional expression of bkt2 than on ipi-1 ipi-2, psy and pds (>4-fold up-regulation). However, GA40 (40 mg/L of GA₃) induced more transcriptional expression of ipi-2, psy and bkt2 than both ipi-1 and pds. The expression of lyc, crtR-B and crtO for astaxanthin biosynthesis was not affected by GA₃ in H. piuvialis.In the presence of GA₃, astaxanthin biosynthesis genes of ipi-1, pds and bkt2 were up-regulated at transcriptional level, psy at post-transcriptional level, whereas ipi-2 was up-regulated at both levels. The study could potentially lead to a scale application of exogenous GA₃ in astaxanthin production with H. pluvialis just like GAsperform in increasing crops production and it would provide new insight about the multifunctional roles of carotenogenesis in response to GA₃. Page(s): 548-553

Inhibitory effect of Piper betle Linn. leaf extract on protein glycation - Quantification and characterization of the antiglycation components

2013-12-01T00:00:00Z

Title: Inhibitory effect of Piper betle Linn. leaf extract on protein glycation - Quantification and characterization of the antiglycation components Authors: Bhattacherjee, Abhishek; Chakraborti, Abhay Sankar Abstract: Piper betle Linn. is a Pan-Asiatic plant having several beneficial properties. Protein glycation and advanced glycation end products (AGEs) formation are associated with different pathophysiological conditions, including diabetes mellitus. Our study aims to find the effect of methanolic extract of P. betle leaves on in vitro protein glycation in bovine serum albumin (BSA)-glucose model. The extract inhibits glucose-induced glycation, thiol group modification and carbonyl formation in BSA in dose-dependent manner. It inhibits different stages of protein glycation, as demonstrated by using glycation models: hemoglobin-d-gluconolactone (for early stage, Amadori product formation), BSA-methylglyoxal (for middle stage, formation of oxidative cleavage products) and BSA-glucose (for last stage, formation of AGEs) systems. Several phenolic compounds are isolated from the extract. Considering their relative amounts present in the extract, rutin appears to be the most active antiglycating agent. The extract of P. betle leaf may thus have beneficial effect in preventing protein glycation and associated complications in pathological conditions. Page(s): 529-536

NOPR Community:

Purification and characterization of structural and functional properties of two lectins from a marine sponge Spheciospongia vesparia

Puriﬁcation and characterization of 6-phosphogluconate dehydrogenase (6-PGD) from grass carp (Ctenopharyngodon idella) hepatopancreas

Carotenoid genes transcriptional regulation for astaxanthin accumulation in fresh water unicellular alga Haematococcus pluvialis by gibberellin A3 (GA3)

Inhibitory effect of Piper betle Linn. leaf extract on protein glycation - Quantification and characterization of the antiglycation components

Carotenoid genes transcriptional regulation for astaxanthin accumulation in fresh water unicellular alga Haematococcus pluvialis by gibberellin A3 (GA₃)