Purification and Characterization of a β-Glucosidase Specific for 2,4-Dihydroxy- 7-methoxy-1,4-benzoxazin-3-one (DIMBOA) Glucoside in Maize

1999 ◽  
Vol 54 (3-4) ◽  
pp. 181-185 ◽  
Author(s):  
Akira Oikawa ◽  
Kenkichi Ebisui ◽  
Masayuki Sue ◽  
Atsushi Ishihara ◽  
Hajime Iwamura
Keyword(s):  
Zea Mays ◽  
Ion Exchange ◽  
Cation Exchange ◽  
Hydroxamic Acid ◽  
Zea Mays L ◽  
Exchange Chromatography ◽  
Cation Exchange Chromatography ◽  
Purification And Characterization ◽  
Apparent Homogeneity

Occurrence and properties of hydroxamic acid glucoside glucosidase were investigated in 10-day-old, autotrophic maize (Zea mays L.) in which 2,4-dihydroxy-7-methoxy-1,4- benzoxazin-3-one glucoside (DIMBOA-G) is a major benzoxazinone component. Crude extracts of both leaves and roots showed glucosidase activity for both DIMBOA-G and 2,4- dihydroxy-1,4-benzoxazin-3-one glucoside (DIBOA-G). A cation-exchange chromatography after cryoprecipitation of the extract from leaves gave a peak with both activities, and further purification by ion-exchange and hydroxyapatite chromatography gave a fraction with an apparent homogeneity, the purification being 560 fold. The Km values (mᴍ) of the purified glucosidase were 0.16 for DIMBOA-G, 0.68 for DIBOA-G and 2.96 for p-nitrophenyl-β-ᴅ-glucopyranoside. The activity on salicin and esculin was too low to be detected. The data indicate that a glucosidase specific for DIMBOA-G comes into contact with constitutive benzoxazinone glucosides producing defensive aglycone when plants are damaged by microbial or insect attacks.

Occurrence of 2,4-Dihydroxy-7-methoxy-1,4-benzoxazin-3-one (DIMBOA) and a β-Glucosidase Specific for Its Glucoside in Maize Seedlings

1998 ◽  
Vol 53 (9-10) ◽  
pp. 793-798 ◽  
Author(s):  
Kenkichi Ebisui ◽  
Atsushi Ishihara ◽  
Nobuhiro Hirai ◽  
Hajime Iwamura
Keyword(s):  
Zea Mays ◽  
Zea Mays L ◽  
Gel Filtration ◽  
Exchange Chromatography ◽  
Autotrophic Growth ◽  
Cation Exchange Chromatography ◽  
Maize Seedlings ◽  
Fresh Weight ◽  
Apparent Homogeneity ◽  
Defense Compound

2,4-Dihydroxy-7-methoxy-1,4-benzoxazin-3-one (DIMBOA) and its glucoside, DIMBOA - G, appeared concurrently with germination in maize (Zea mays L.), and their concentrations per fresh weight reached a maximum 24-36 hr after germination. The aglycone then decreased to disappear as the plants began autotrophic growth, and the glucoside to a concentration half that of the maximum. The level of DIMBOA-G was always higher than that of the aglycone. A β-glucosidase activity which hydrolyzes DIMBOA-G to DIMBOA and glucose was detected in crude enzyme extracts from the seedlings. Its activity per fresh weight varied concurrently with the occurrence of DIMBOA. The purification was performed to an apparent homogeneity by cryoprecipitation followed by cation exchange chromatography and gel filtration. The Km value for DIMBOA-G was 0.07 mᴍ, whereas that for DIBOA-G, that was substantially absent in the seedlings of the cultivar studied, was 0.52 mᴍ. The activity on salicin and esculin was too low to be detected. The set of results suggested that free DIMBOA occurs constitutively or in a programmed manner as defense compound at a seedling stage of growth, and a specific glucosidase is involved in this process.

Characterization of the Acidic Species of a Monoclonal Antibody Using Weak Cation Exchange Chromatography and LC-MS

2015 ◽  
Vol 87 (17) ◽  
pp. 9084-9092 ◽  
Author(s):  
Gomathinayagam Ponniah ◽  
Adriana Kita ◽  
Christine Nowak ◽  
Alyssa Neill ◽  
Yekaterina Kori ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Cation Exchange ◽  
Exchange Chromatography ◽  
Cation Exchange Chromatography ◽  
Weak Cation Exchange

scholarly journals Purification and characterization of the heat-stable serine proteinase from Thermomonospora fusca YX

1987 ◽  
Vol 246 (2) ◽  
pp. 511-517 ◽  
Author(s):  
T W Gusek ◽  
J E Kinsella
Keyword(s):  
Thermal Stability ◽  
Ionic Strength ◽  
Serine Proteinase ◽  
Maximum Activity ◽  
Exchange Chromatography ◽  
Cation Exchange Chromatography ◽  
Purification And Characterization ◽  
Thermomonospora Fusca ◽  
Heat Stable

The proteinase secreted from Thermomonospora fusca YX grown on cellulose was purified by (NH4)2SO4 fractionation and cation-exchange chromatography. The isolated proteinase readily hydrolysed several proteins and demonstrated activity towards casein from 35 to 95 degrees C (at pH 8.0) with maximum activity at 80 degrees C. It exhibited broad pH and ionic-strength optima centered at pH 9.0 and 0.2 M-NaCl respectively, and it retained high activity in the presence of 2% (w/v) SDS, 20 mM-dithiothreitol and 1.0 M-NaCl. The proteinase, which was fully inhibited by phenylmethanesulphonyl fluoride, had an Mr of 14,500 and an isoelectric point at 9.21. A measurement of proteinase thermal stability demonstrated a T50% (15 min) of 85 degrees C at pH 4.5.

scholarly journals Purification and characterization of an α2-macroglobulin-like proteinase inhibitor from plasma of the crayfish Pacifastacus leniusculus

1988 ◽  
Vol 255 (3) ◽  
pp. 801-806 ◽  
Author(s):  
H G Hergenhahn ◽  
M Hall ◽  
K Söderhäll
Keyword(s):  
Proteinase Inhibitor ◽  
Hydrophobic Interaction Chromatography ◽  
Acid Precipitation ◽  
Anion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Pacifastacus Leniusculus ◽  
Purification And Characterization ◽  
Apparent Homogeneity ◽  
Α2 Macroglobulin

An alpha 2-macroglobulin (alpha 2M)-like proteinase inhibitor from plasma of the crayfish Pacifastacus leniusculus was purified to apparent homogeneity by acid precipitation, hydrophobic interaction chromatography, affinity chromatography on concanavalin A-Sepharose and anion-exchange chromatography. The subunit Mr is about 190,000. Pore-size-limit electrophoresis proved the native protein to be a dimer. The purified protein resembled vertebrate alpha 2 Ms in that it protected trypsin from inhibition by soyabean trypsin inhibitor, and in its sensitivity to methylamine treatment. Methylamine also prevented the protein from being autolytically cleaved into Mr 60,000 and 140,000 fragments when subjected to heat treatment. The amino acid composition showed similarities with both human alpha 2 M and an alpha 2 M-like protein from the arthropod Limulus polyphemus. These data indicate that this Pacifastacus alpha 2M-like protein (P alpha 2M) may be a distantly related homologue of vertebrate alpha 2Ms.

scholarly journals Purification and Characterization of Porin from Corn (Zea mays L.) Mitochondria

1993 ◽  
Vol 102 (2) ◽  
pp. 615-621 ◽  
Author(s):  
J. A. Aljamal ◽  
G. Genchi ◽  
V. De Pinto ◽  
L. Stefanizzi ◽  
A. De Santis ◽  
...  
Keyword(s):  
Zea Mays ◽  
Zea Mays L ◽  
Purification And Characterization

scholarly journals Ion-Exclusion/Cation-Exchange Chromatography Using Dual-Ion-Exchange Groups for Simultaneous Determination of Inorganic Ionic Nutrients in Fertilizer Solution Samples for the Management of Hydroponic Culture

Agronomy ◽  
2021 ◽  
Vol 11 (9) ◽  
pp. 1847
Author(s):  
Daisuke Kozaki ◽  
Yuki Sago ◽  
Taku Fujiwara ◽  
Masanobu Mori ◽  
Chihiro Kubono ◽  
...  
Keyword(s):  
Ion Exchange ◽  
Cation Exchange ◽  
Simultaneous Determination ◽  
Matrix Effects ◽  
Exchange Chromatography ◽  
Hydroponic Culture ◽  
Cation Exchange Chromatography ◽  
Ion Exclusion ◽  
Fertilizer Solution

In this study, ion-exclusion/cation-exchange chromatography (IEC/CEC) using dual-ion-exchange groups (carboxy and sulfo groups) for the simultaneous determination of anions (SO42−, Cl−, NO3−, and HPO42−) and cations (Na+, NH4+, K+, Mg2+, and Ca2+) was developed. By using the combination of dual-ion-exchange groups, simultaneous separation of inorganic ions with HPO42− was achieved that was impossible by the conventional IEC/CEC based on the single-ion-exchange group (carboxy group). This method was applied to the monitoring of inorganic ionic nutrients in fertilizer solution samples in hydroponic culture. As a result, a higher peak resolution of inorganic anions and cations with phosphate ion using IEC/CEC with dual-ion-exchange groups was achieved in the absence of matrix effects. In addition, the developed method helps to understand the behavior of ionic nutrients in fertilizer solution during hydroponic cultivation and is potentially useful for the individual fertilization of ionic nutrients.

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