scholarly journals Enzymic synthesis of sym-homospermidine in Lathyrus sativus (grass pea) seedlings

1980 ◽  
Vol 190 (2) ◽  
pp. 461-464 ◽  
Author(s):  
K S Srivenugopal ◽  
P R Adiga
Keyword(s):  
Enzyme Activity ◽  
Affinity Chromatography ◽  
Specific Activity ◽  
Lathyrus Sativus ◽  
Santalum Album ◽  
Grass Pea ◽  
Enzymic Synthesis ◽  
Crude Extracts ◽  
Pea Seedlings ◽  
Blue Sepharose

An enzyme catalysing the synthesis of sym-homospermidine from putrescine and NAD+ with concomitant liberation of NH3 was purified 100-fold from Lathyrus sativus (grass pea) seedlings by affinity chromatography on Blue Sepharose. This thiol enzyme had an apparent mol.wt. of 75000 and exhibited Michelis-Menten kinetics with Km 3.0mM for putrescine. The same enzyme activity could also be demonstrated in the crude extracts of sandal (Santalum album) leaves, but with a specific activity 15-fold greater than that in L. sativus seedlings.

Abscisic acid promotes accumulation of toxin ODAP in relation to free spermine level in grass pea seedlings (Lathyrus sativus L.)

2006 ◽  
Vol 44 (2-3) ◽  
pp. 161-169 ◽  
Author(s):  
You-Cai Xiong ◽  
Geng-Mei Xing ◽  
Feng-Min Li ◽  
Shao-Ming Wang ◽  
Xian-Wei Fan ◽  
...  
Keyword(s):  
Abscisic Acid ◽  
Lathyrus Sativus ◽  
Grass Pea ◽  
Pea Seedlings ◽  
Lathyrus Sativus L

Correlation of drought resistance in grass pea (Lathyrus sativus) with reactive oxygen species scavenging and osmotic adjustment

Biologia ◽  
2013 ◽  
Vol 68 (2) ◽  
Author(s):  
Jinglong Jiang ◽  
Miao Su ◽  
Yueru Chen ◽  
Nan Gao ◽  
Chengjin Jiao ◽  
...  
Keyword(s):  
Soluble Sugars ◽  
Lathyrus Sativus ◽  
Transcriptional Level ◽  
Grass Pea ◽  
Stress Injury ◽  
Garden Pea ◽  
Legume Crop ◽  
Drought Tolerant ◽  
Pea Seedlings ◽  
Lathyrus Sativus L

AbstractGrass pea (Lathyrus sativus L.), a legume crop in arid and semi-arid areas, is widely acknowledged as highly drought tolerant. We report here an analysis of grass pea and garden pea seedlings stressed with 20% polyethylene glycol 6000 (PEG) for five days. While leaf margins of grass pea curled inward after PEG stress, leaves of pea failed to display this trait. PEG inhibited the growth of grass pea less than that of pea. Hydrogen peroxide (H2O2) and malondialdehyde (MDA) accumulation increased in pea more than in grass pea. Greater accumulation of proline and soluble sugars alleviated osmotic stress injury to grass pea compared with pea. Moreover, PEG caused a significantly greater increase of superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX) and glutathione peroxidase (GPX) activities in grass pea compared to pea. These enzymes showed obvious up-regulation at the transcriptional level in grass pea leaves. Together, these data suggest that the accumulation of osmoprotectants and the improvement of oxidation resistance resulted in the higher drought tolerance of grass pea compared to pea.

scholarly journals Characterization of angiotensin-converting enzyme in canine testis

Reproduction ◽  
2001 ◽  
pp. 139-146 ◽  
Author(s):  
K Sabeur ◽  
AT Vo ◽  
BA Ball
Keyword(s):  
Enzyme Activity ◽  
Angiotensin Converting Enzyme ◽  
Affinity Chromatography ◽  
Ace Inhibitor ◽  
Specific Activity ◽  
Normal Activity ◽  
Seminiferous Tubules ◽  
Affinity Column ◽  
Converting Enzyme ◽  
Ace Activity

The aim of this study was to characterize angiotensin-converting enzyme (ACE) in canine testis. Detergent-extracted canine testes were sonicated in the presence of protease inhibitors and purified on an affinity column with the ACE inhibitor, lisinopril, as an affinity ligand for ACE. The fractions recovered were assessed for ACE enzyme activity via an enzyme kinetic microplate assay (at 330 nm) based on the hydrolysis of Fa-Phe-Gly-Gly (FAPGG) at pH 7.5 during an 8 min incubation. The specific activity of ACE in the starting testicular extracts was 3.53 +/- 0.99 mU mg(-1) protein with a 1588 times enrichment in ACE activity after lisinopril affinity chromatography (4239 +/- 2600 mU mg(-1) protein). The recovery efficiency of ACE after lisinopril affinity chromatography was 71.2%. The ACE activity in the detergent extracts and the purified fractions was inhibited significantly by 10 micromol captopril l(-1), a specific ACE inhibitor, and was restored to 88% of normal activity by the addition of the thiol-alkylating agent N-ethylmaleimide (0.5 mmol l(-1)) in the detergent extracts and the purified fractions incubated with captopril. The treatment of testicular extracts with 10 mmol EDTA l(-1) reduced the ACE activity significantly (5.40 +/- 1.26 versus 0.58 +/- 0.23 mU mg(-1)). The ACE activity was restored fully in the presence of zinc (5.28 +/- 0.70 mU mg(-1)). The anti-ACE antibody (raised against a 70 kDa protein from the periacrosomal plasma membrane of equine spermatozoa) recognized a 65-70 kDa protein in the detergent-extracted testes as well as in the affinity-purified fractions. This antibody also recognized a protein of similar molecular mass in ejaculated spermatozoa. ACE was localized in the periacrosomal area of the ejaculated spermatozoa and in spermatids in the seminiferous tubules. The results of this study demonstrate that ACE is present in canine testis and retains its enzyme activity after purification with lisinopril affinity chromatography. Activity of canine ACE is inhibited by captopril and EDTA and is restored in the presence of N-ethylmaleimide and zinc.

Accumulation pattern of toxin β-ODAP during lifespan and effect of nutrient elements on β-ODAP content in Lathyrus sativus seedlings

2006 ◽  
Vol 144 (4) ◽  
pp. 369-375 ◽  
Author(s):  
CHENG-JIN JIAO ◽  
QUAN-LE XU ◽  
CHONG-YING WANG ◽  
FENG-MIN LI ◽  
ZHI-XIAO LI ◽  
...  
Keyword(s):  
High Performance ◽  
Field Conditions ◽  
Lathyrus Sativus ◽  
Nutrient Elements ◽  
Grass Pea ◽  
Nitrogen And Phosphorus ◽  
Accumulation Pattern ◽  
Nitrogenous Compounds ◽  
Pea Seedlings ◽  
Young Leaves

In order to assess the influence of nutrient elements on the accumulation of β-N-oxalyl-L-α, β-diaminopropionic acid (β-ODAP; the probable cause of lathyrism) in Lathyrus sativus L. (grass pea), it was first examined under field conditions during the lifespan of a grass pea plant using high-performance liquid chromatography (HPLC). β-ODAP mainly accumulated in young seedlings, in developing and mature seeds and in young leaves, especially in young seedlings. In contrast, all mature leaves, roots, and stems showed a low level of β-ODAP. The β-ODAP accumulation pattern in seedlings grown in a nutrient-deficient solution was highest (3·57 mg/g) in shoots at 7 d growing in the nitrogen-deficient solution and higher compared to the control (2·31 mg/g) in zinc-, calcium-, phosphorus- and molybdenum-deficient shoots (P<0·05). The contents of β-ODAP in seedlings growing in other element-deficient solutions were similar to controls. When the content of β-ODAP in grass pea seedlings fertilized with different forms of organic nitrogen was assayed the results indicated that amino acids such as glutamine and serine, as well as nucleotide nitrogen, all significantly enhanced the accumulation of β-ODAP in young seedlings relative to controls (P<0·05). Taken together, these data suggest that β-ODAP accumulation in grass pea might be related to the level of total free nitrogenous compounds and that nitrogen and phosphate may be the crucial nutrient factors influencing β-ODAP content under field conditions. Thus, the application of appropriate nitrogen and phosphorus fertilizers to the soil could decrease the content of β-ODAP in the seeds and leaves of grass pea.

Separation of Pinusbanksiana Lamb. glyceraldehyde 3-phosphate (NAD) dehydrogenase from phenolic-based pigments with affinity chromatography

1982 ◽  
Vol 12 (4) ◽  
pp. 1035-1038
Author(s):  
Bruce E. Haissig
Keyword(s):  
Enzyme Activity ◽  
Affinity Chromatography ◽  
Electrophoretic Mobility ◽  
Spectrophotometric Assay ◽  
Disc Electrophoresis ◽  
Enzyme Complex ◽  
Crude Extracts ◽  
Ungerminated Seed ◽  
Catalytically Active ◽  
The Individual

Glyceraldehyde 3-phosphate (NAD) dehydrogenase was extracted from Pinusbanksiana Lamb, seed and from seedlings up to 12 days old. Enzyme activity in crude extracts of seedlings increased markedly from days 2–8 and then decreased slightly, as indicated by spectrophotometric assay. In contrast, staining of acrylamide gels after disc electrophoresis indicated that catalytically active isozymes were not present in crude extracts at and after day 6. Phenolics greatly increased in extracts from days 0–10 and polymerization of these phenolics apparently lead to binding of the resulting phenolic-based pigment to enzyme. The pigment–enzyme complex demonstrated greatly enhanced electrophoretic movement, in comparison with the individual isozymes, which caused frontal migration of the collective isozymes. Affinity chromatography restored electrophoretic mobility of isozymes equal to that obtained with enzyme from ungerminated seed. Enzyme was inseparable from pigment by several other methods.

Mixed Mode Chromatography: A Novel Way Toward New Selectivity

2018 ◽  
Vol 20 (1) ◽  
pp. 14-21 ◽  
Author(s):  
Xavier Santarelli ◽  
Charlotte Cabanne
Keyword(s):  
Salt Tolerance ◽  
Affinity Chromatography ◽  
Mixed Mode ◽  
Ionic Groups ◽  
Crude Extracts ◽  
The Way

Mixed mode chromatography offers a diversity of ligands, each providing a new selectivity. This allows the design of novel purification processes with reduced column steps. Structure of ligands is based on both hydrophobic and ionic groups. Thanks to its salt tolerance, crude extracts or post-IEX samples can be loaded directly without conditioning. The selectivity could be enhanced by modulating elution parameters or by using additives. More importantly, mixed mode chromatography could be as effective as affinity chromatography for mAb purification processes. Mixed mode chromatography opens the way to short and economical processes.

scholarly journals Study of Extraction and Enzymatic Properties of Cell-Envelope Proteinases from a Novel Wild Lactobacillus plantarum LP69

Catalysts ◽  
2018 ◽  
Vol 8 (8) ◽  
pp. 325 ◽  
Author(s):  
He Chen ◽  
Jie Huang ◽  
Binyun Cao ◽  
Li Chen ◽  
Na Song ◽  
...  
Keyword(s):  
Enzyme Activity ◽  
Lactobacillus Plantarum ◽  
Industrial Development ◽  
Extraction Efficiency ◽  
Specific Activity ◽  
Cell Envelope ◽  
Serine Proteinase ◽  
Kinetic Studies ◽  
Predicted Values ◽  
Hydrolyze Whey Protein

Lactobacilli cell-envelope proteinases (CEPs) have been widely used in the development of new streams of blockbuster nutraceuticals because of numerous biopharmaceutical potentials; thus, the development of viable methods for CEP extraction and the improvement of extraction efficiency will promote their full-scale application. In this study, CEP from a novel wild Lactobacillus plantarum LP69 was released from cells by incubating in calcium-free buffer. The extraction conditions of CEP were optimized by response surface methodology with the enzyme activity and specific activity as the detective marker. The optimal extraction conditions were: time of 80 min, temperature of 39 °C and buffer pH of 6.5. Under these conditions, enzyme activity and specific activity were (23.94 ± 0.86) U/mL and (1.37 ± 0.03) U/mg, respectively, which were well matched with the predicted values (22.12 U/mL and 1.36 U/mg). Optimal activity of the crude CEP occurred at pH 8.0 and 40 °C. It is a metallopeptidase, activated by Ca2+, inhibited by Zn2+ and ethylene-diamine-tetra-acetic acid, and a serine proteinase which is inhibited by phenylmethylsulfonyl fluoride. Kinetic studies showed that CEP from LP69 could hydrolyze whey protein, lactoglobulin and casein. Our study improves the extraction efficiency of CEPs from LP69, providing the reference for their industrial development.

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