Purification of damson plum polyphenol oxidase by affinity chromatography and investigation of metal effects on enzyme activity

2022 ◽  
pp. 1-16
Author(s):  
Selinnur Yıldız ◽  
Çiğdem Bilen ◽  
Emine Karakuş
Keyword(s):  
Enzyme Activity ◽  
Affinity Chromatography ◽  
Polyphenol Oxidase

scholarly journals Enzyme activity in assessing the effects of fumigation with soil smoke

2021 ◽  
Vol 4 (46) ◽  
pp. 26-26
Author(s):  
Alexander Saakian ◽  
◽  
Keyword(s):  
Thermal Decomposition ◽  
Enzyme Activity ◽  
Polyphenol Oxidase ◽  
Soil Enzymes ◽  
Combustion Products ◽  
Wood Chips ◽  
Ordinary Chernozem ◽  
Plant Origin ◽  
Plant Materials ◽  
Catalase Peroxidase

Abstract As a result of fires, in addition to the fire itself and high temperatures, smoke from combustion products has a significant effect on the biota. The aim of the work was to assess the effect of fumigation with combustion products of plant origin on the biology activity of ordinary chernozem. In a series of model experiments, the reaction of soil enzymes (catalase, peroxidase, polyphenol oxidase, invertase, urease, phosphatase) to the smoke of the studied soil with products of thermal decomposition of plant materials (foliage, needles, straw, wood chips) is shown. A significant decrease in the enzyme activity of the studied enzymes was revealed in the range from 7% to 33%, depending on the time spent under the smoke of chernozem (15–120 minutes). The highest sensitivity to fumigation was noted for enzymes of the class of oxidoreductases: catalase, polyphenoloxidase, and peroxidase. Thus, a significant sensitivity and information content of the indicators of the enzyme activity of soils on the effect of smoke has been established, which can be used in monitoring the consequences of fires. Keywords: PYROGENIC EFFECTS, WILDFIRE, COMBUSTION PRODUCTS, CHERNOZEM, BIOLOGY ACTIVITY

Single-step purification of urease by affinity chromatography

1974 ◽  
Vol 20 (4) ◽  
pp. 623-630 ◽  
Author(s):  
Bassanio L. Wong ◽  
Charles R. Shobe
Keyword(s):  
Enzyme Activity ◽  
Affinity Chromatography ◽  
Single Step ◽  
Jack Bean Urease ◽  
Jack Bean ◽  
Specific Activities ◽  
Single Step Purification

Single-step purification of urease (urea aminohydrolase; EC. 3.5.1.5) from cell-free extracts of Proteus morganii and from partially purified preparations of jack bean urease were achieved in less than 90 min by affinity chromatography on hydroxyurea-substituted beaded agarose columns. The specific activities of the purified enzymes were as high as, or higher than, those reported by other authors for urease preparations obtained by conventional techniques. In addition, yields of enzyme activity were routinely 6 to 100 times greater than recoveries previously reported for this enzyme.

scholarly journals Identification of rat liver phosphatidylinositol synthase as a 21 kDa protein

1994 ◽  
Vol 304 (1) ◽  
pp. 301-305 ◽  
Author(s):  
M E Monaco ◽  
M Feldman ◽  
D L Kleinberg
Keyword(s):  
Heat Treatment ◽  
Enzyme Activity ◽  
Affinity Chromatography ◽  
Molecular Mass ◽  
Rat Liver ◽  
Protein Band ◽  
Absolute Requirement ◽  
Sds Page ◽  
Sepharose 4B ◽  
Post Mitochondrial Supernatant

Substantial purification of rat liver phosphatidylinositol (PtdIns) synthase has been achieved by a combination of Hecameg extraction, heat treatment, affinity chromatography and chromatography on PBE-94. The activity chromatographs as a single peak which has an apparent molecular mass between 150 and 200 kDa on Sepharose 4B. When analysed by SDS/PAGE, two major bands are seen. The enzyme activity is correlated with a protein band of 21 kDa. A second band, at 51 kDa, is eluted from a PBE-94 column slightly ahead of the activity. Manganese is an absolute requirement for stabilization of activity in the presence of detergent. The effect of manganese is optimal at 0.5 mM; magnesium at a concentration of 10 mM is only minimally effective. Substrate Kms are 1.3 mM and 9.5 microM for inositol and CDP-diacylglycerol respectively. The activity eluting from the PBE-94 column is purified 5000-fold over the post-mitochondrial supernatant.

scholarly journals Enzyme activity in wheat breeding lines derived from matings of low polyphenol oxidase parents

Euphytica ◽  
2012 ◽  
Vol 190 (1) ◽  
pp. 65-73 ◽  
Author(s):  
Somrudee Nilthong ◽  
R. A. Graybosch ◽  
P. S. Baenziger
Keyword(s):  
Enzyme Activity ◽  
Polyphenol Oxidase ◽  
Wheat Breeding ◽  
Breeding Lines

scholarly journals Characterization of glutamine synthetase from the ammonium-excreting strain HM053 of Azospirillum brasilense

2022 ◽  
Vol 82 ◽  
Author(s):  
Fernanda Ghenov ◽  
Edileusa Cristina Marques Gerhardt ◽  
Luciano Fernandes Huergo ◽  
Fabio Oliveira Pedrosa ◽  
Roseli Wassem ◽  
...  
Keyword(s):  
Enzyme Activity ◽  
Glutamine Synthetase ◽  
Affinity Chromatography ◽  
Azospirillum Brasilense ◽  
Nitrogen Assimilation ◽  
Atp Hydrolysis ◽  
Nitrogen Fixing ◽  
Wild Type ◽  
E Coli

Abstract Glutamine synthetase (GS), encoded by glnA, catalyzes the conversion of L-glutamate and ammonium to L-glutamine. This ATP hydrolysis driven process is the main nitrogen assimilation pathway in the nitrogen-fixing bacterium Azospirillum brasilense. The A. brasilense strain HM053 has poor GS activity and leaks ammonium into the medium under nitrogen fixing conditions. In this work, the glnA genes of the wild type and HM053 strains were cloned into pET28a, sequenced and overexpressed in E. coli. The GS enzyme was purified by affinity chromatography and characterized. The GS of HM053 strain carries a P347L substitution, which results in low enzyme activity and rendered the enzyme insensitive to adenylylation by the adenilyltransferase GlnE.

scholarly journals Modification of Cu2+ in polyphenol oxidase extract from purple eggplant for phenol degradation in coal wastewater treatment

2021 ◽  
Vol 882 (1) ◽  
pp. 012071
Author(s):  
A Murniati ◽  
B Buchari ◽  
S Gandasasmita ◽  
Z Nurachman ◽  
VA Kusumaningtiyas ◽  
...  
Keyword(s):  
Enzyme Activity ◽  
Polyphenol Oxidase ◽  
Coal Gasification ◽  
Initial Rate ◽  
Phenol Degradation ◽  
Solanum Melongena ◽  
Liquid Waste ◽  
Phenol Biodegradation ◽  
Gasification Process ◽  
Phenol Determination

Abstract Cracking coal-forming organic compounds during the gasification process produces liquid waste containing phenolic compounds that require special handling based on their toxicity. As one of the components, there is liquid waste resulting from the coal gasification process. The purpose of this research was to study the activity of polyphenol oxidase (PPO) on phenol after the addition of Cu2+ to purple eggplant (Solanum melongena L.) extract and its potential to work more effectively in phenol biodegradation for coal wastewater containing phenol. Enzyme activity and phenol determination were carried out spectrophotometrically. The results showed PPO activity of 25.90-38.10 U/mL; 4.0 mM phenol and the activity of PPO-Cu2+ was 21.58-46.32 U/mL; 2-10 mM CuSO4; 2.0-4.0 mM phenol. Based on Michaelis Menten’s graph, the initial rate of PPO-Cu2+ was 0.015 mM/min and the initial rate of PPO was 0.15 mM/min using 2 mM phenol as a substrate. Lineweaver-Burk’s graph shows the KM of PPO-Cu2+ = 6.92 mM, which is lower than KM of PPO = 13.05 mM. Its means that the phenol response has a higher affinity for PPO-Cu2+ than PPO. The application of PPO-Cu2+ in purple eggplant extract works effectively as much as 46.7% for artificial coal liquid waste containing phenol.

scholarly journals Role of Polyphenol Oxidase, Peroxidase, Phenylalanine Ammonia Lyase, Chlorogenic Acid, and Total Soluble Phenols in Resistance of Potatoes to Soft Rot

Plant Disease ◽  
2012 ◽  
Vol 96 (2) ◽  
pp. 186-192 ◽  
Author(s):  
Elizabeth Ngadze ◽  
David Icishahayo ◽  
Teresa A. Coutinho ◽  
Jacquie E. van der Waals
Keyword(s):  
Enzyme Activity ◽  
Chlorogenic Acid ◽  
Polyphenol Oxidase ◽  
Phenylalanine Ammonia Lyase ◽  
Soft Rot ◽  
Defense Responses ◽  
Bacterial Invasion ◽  
Pal Activity ◽  
Soluble Phenols

Pectobacterium atrosepticum, P. carotovorum subsp. brasiliensis, and Dickeya spp. cause soft rot of potato (Solanum tuberosum) worldwide. Plants respond to bacterial invasion by activating defense responses associated with accumulation of several enzymes and inhibitors, which prevent pathogen infection. This study focused on the role of polyphenol oxidase (PPO), peroxidase (POD), phenylalanine ammonia lyase (PAL), chlorogenic acid, and total soluble phenols in imparting resistance to soft rot pathogens. Seven and 11 varieties grown by farmers in South Africa and Zimbabwe, respectively, were used in the study. The results showed significantly higher (P < 0.001) enzyme activity of PPO and PAL as well as higher concentrations of chlorogenic acid and total soluble phenols in Vanderplank, Pentland Dell, M69/11, Romano, M59/20, and MondialZw. PAL activity increased significantly with time in all varieties, and the highest activity was recorded 8 h after cutting. The resistance of the varieties was correlated with high PPO and PAL enzyme activity as well as increased concentrations of chlorogenic acid and total soluble phenols. PPO, POD, and PAL activities increased significantly in wounded and inoculated tubers. These findings show that PAL, PPO, POD, chlorogenic acid, and total soluble phenols play a role in imparting resistance to potato soft rot infection.

Centrifugal analyzer used for enzyme immunoassay of progesterone and choriomammotropin.

1983 ◽  
Vol 29 (2) ◽  
pp. 302-304 ◽  
Author(s):  
B Terouanne ◽  
J Marchand ◽  
C Calzolari ◽  
J Monnier ◽  
J C Nicolas ◽  
...  
Keyword(s):  
Enzyme Activity ◽  
Affinity Chromatography ◽  
Enzyme Immunoassay ◽  
Gel Filtration ◽  
Placental Lactogen ◽  
Human Placental Lactogen ◽  
Rapid Separation ◽  
Enzyme Conjugate ◽  
Enzyme Label ◽  
Free Antigen

Abstract Recently we developed an enzyme immunoassay involving the use of steroid delta-isomerase (EC 5.3.3.1) as enzyme label and exclusion-affinity chromatography for rapid separation of free antigen-enzyme conjugate that bound to antibodies (J. Immunol. Methods 35: 267-284, 1980). Here we describe an automated version of this procedure, for immunoassay of progesterone and choriomammotropin (human placental lactogen) in serum with the use of a centrifugal analyzer. After incubation, suitable dilutions of sera or extract plus antiserum, conjugate, and double antibody were filtered on an estradiol affinity gel-filtration column; the enzyme activity of the filtrates was determined with the centrifugal analyzer. Results correlate well with those obtained by radioimmunoassay: r (progesterone) = 0.980, r (choriomammotropin) = 0.940. The within-run and between-run precision, specificity, sensitivity, accuracy, and speed of this system make it a useful tool for immunoassay.

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