EJPAU 2008. Sempruch C. , Ciepiela A. , Sprawka I. , Chrzanowski G. PURIFICATION AND SOME PHYSICOCHEMICAL PROPERTIES OF NITRATE REDUCTASE ISOLATED FROM WINTER TRITICALE SEEDLINGS

Electronic Journal of Polish Agricultural Universities (EJPAU) founded by all Polish Agriculture Universities presents original papers and review articles relevant to all aspects of agricultural sciences. It is target for persons working both in science and industry,regulatory agencies or teaching in agricultural sector. Covered by IFIS Publishing (Food Science and Technology Abstracts), ELSEVIER Science - Food Science and Technology Program, CAS USA (Chemical Abstracts), CABI Publishing UK and ALPSP (Association of Learned and Professional Society Publisher - full membership). Presented in the Master List of Thomson ISI.

2008
Volume 11
Issue 1/volume11

Topic:

Biology

ELECTRONIC
JOURNAL OF
POLISH
AGRICULTURAL
UNIVERSITIES

Sempruch C. , Ciepiela A. , Sprawka I. , Chrzanowski G. 2008. PURIFICATION AND SOME PHYSICOCHEMICAL PROPERTIES OF NITRATE REDUCTASE ISOLATED FROM WINTER TRITICALE SEEDLINGS, EJPAU 11(1/volume11), #14.
Available Online: http://www.ejpau.media.pl/volume11/issue1/volume11/abs-14.html

PURIFICATION AND SOME PHYSICOCHEMICAL PROPERTIES OF NITRATE REDUCTASE ISOLATED FROM WINTER TRITICALE SEEDLINGS

Cezary Sempruch, Antoni P. Ciepiela, Iwona Sprawka, Grzegorz Chrzanowski
Department of Biochemistry and Molecular Biology, University of Podlasie, Siedlce, Poland

ABSTRACT

Nitrate reductase (NR, EC 1.6.6.1-3) is a key enzyme of assimilative reduction of nitrate in plant tissues. NRs present in higher plants are differed in structure and physicochemical properties. In literature there are information concerning properties of the enzymes isolated from tissues such cereals as wheat, maize, rice and barley, but not triticale. Because of that, the aim of the research was to purify nitrate reductase isolated from the leaves of seven-day-old seedlings of winter triticale cultivar Moreno and to analyze its physicochemical properties.

The research results showed that NR obtained from the winter triticale seedlings is a homogeneous protein fraction of the molecular weight of approximately 190000 Da. The absorbency of the purified enzyme reached its maximum values at 244, 463 and 491 nm. NR was specific in relation to NADH₂ as well as to NADPH₂ although it showed lower specific activity in the presence of the latter coenzyme. NR isolated from the seeds of the winter triticale also showed other activities such as MVH (methylviologen) – NR, FMNH₂ – NR, NADH-ferricyanide reductase and NADH-cyt c reductase, which is the evidence of the enzyme partial ability to catalyze reactions other than the reduction of nitrate to nitrite in the presence of NADH₂ or NADPH₂ as the electron donors.

Key words: nitrate reductase, winter triticale, assimilative reduction of nitrate.

Cezary Sempruch
Department of Biochemistry and Molecular Biology, University of Podlasie, Siedlce, Poland
B. Prusa 12, 08-110 Siedlce, Poland
email: cezar@ap.siedlce.pl

Antoni P. Ciepiela
Department of Biochemistry and Molecular Biology, University of Podlasie, Siedlce, Poland
B. Prusa 12, 08-110 Siedlce, Poland

Iwona Sprawka
Department of Biochemistry and Molecular Biology, University of Podlasie, Siedlce, Poland
B. Prusa 12, 08-110 Siedlce, Poland

Grzegorz Chrzanowski
Department of Biochemistry and Molecular Biology, University of Podlasie, Siedlce, Poland
B. Prusa 12, 08-110 Siedlce, Poland
email: grzegorzc@ap.siedlce.pl

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