Purification and stability during storage of phoshoenolpyruvate carboxylase from leaves of Amaranthus hypochondriacus, a NAD-ME type C4 plant

Title:

Purification and stability during storage of phoshoenolpyruvate carboxylase from leaves of Amaranthus hypochondriacus, a NAD-ME type C4 plant

Creator:

Gayathri, J., Parvathi, K., and Raghavendra, A. S.

Identifier:

https://cdk.lib.cas.cz/client/handle/uuid:a9d816b9-39a1-49d4-96e6-c57f1adfa4bc
uuid:a9d816b9-39a1-49d4-96e6-c57f1adfa4bc
issn:0300-3604
doi:10.1023/A:1026739806712

Subject:

DEAE-Sepharose, glucose-6-phosphate, glycerol, hydroxylapaatite, L-malate, phosphate, phosphoenolpyravate, and Seralose-6B

Type:

model:article and TEXT

Format:

bez média and svazek

Description:

A traditional method is reported for purification of phosphoenolpyruvate carboxylase (PEPC; EC 4.1.1.31) from leaves of Amaranthus hypochondriacus L. with a high yield of 50 %, 135-fold purification, and specific activity of 900 mmol kg-1(protein) s-1. PEPC was purified from light-adapted leaves of A. hypochondriacus, involving 40-60 % ammonium sulphate fractionation, followed by chromatography on columns of DEAE-Sepharose, hydroxylapatite (HAP), and Seralose 6-B. The enzyme appeared as a single band on 10 % SDS-PAGE, with a molecular mass of about 100 kDa. Kinetic studies with purified enzyme confirmed the PEPC to be the light-form of the enzyme. Glycerol generally increased the stability of PEPC. The stability and storage of the purified enzyme was studied at temperatures of 4 °C, -20 °C, and liquid nitrogen. PEPC maintained its activity for up to 3 months upon storage with 50 % (v/v) glycerol in liquid nitrogen. and J. Gayathri, K. Parvathi, A. S. Raghavendra.

Language:

Multiple languages

Rights:

http://creativecommons.org/licenses/by-nc-sa/4.0/
policy:public

Coverage:

45-52

Source:

Photosynthetica | 2000 Volume:38 | Number:1

Harvested from:

CDK

Metadata only:

false