Molecular Biology and Genetics |
|
Microbial
Biotechnology |
| Electronic Journal of Biotechnology ISSN: 0717-3458 |
Vol.
12 No. 3, Issue of July 15, 2009 |
| © 2009 by Pontificia Universidad Católica
de Valparaíso -- Chile |
Received September 2, 2008
/ Accepted April 29, 2009 |
| DOI: 10.2225/vol12-issue3-fulltext-14 |
|
Gene expression and characterization of 2-keto-3-deoxy-gluconate kinase,
a key enzyme in the modified Entner-Doudoroff pathway of Serratia marcescens KCTC 2172
Yong-Seok Lee
Department
of Biotechnology
Faculty
of Natural Resources and Life Science
Dong-A
University
Busan,
604-714, Korea
In-Hye
Park
Department
of Biotechnology
Faculty
of Natural Resources and Life Science
Dong-A
University
Busan,
604-714, Korea
Ju-Soon Yoo
Department
of Food Science and Nutrition
Dong-Ju
College
Busan,
Korea
Hae-Sun Kim
Department
of Biotechnology
Dong-A
University
Busan,
604-714, Korea
Soo-Yeol Chung
Department
of Food Science
Dong-Ju
College
Busan,
604-715, Korea
Muni Ramanna GariSubhosh Chandra
Department
of Biotechnology
Dong-A
University
Busan,
604-714, Korea
Yong-Lark Choi*
Department of Biotechnology
Faculty of Natural Resources and
Life Science
Dong-A University 840, Hadan-dong
Saha-gu, Busan, 604-714, Korea
Tel: 82 51 200 6536
Fax: 82 51 200 6536
E-mail: ylchoi@dau.ac.kr
*Corresponding author
Financial support: Research fund of Dong-A University.
Keywords: 2-keto-3-deoxygluconate kinase, carbohydrate kinase, purification, Serratia marcescens KCTC 2172.
Abbreviations: |
GST: Glutatione S-transferase
KDG:
2-keto-3-deoxygluconate
KDGK:
2-keto-3-deoxy-gluconate kinase
KDPG:
2-keto-3-deoxy-6-phophogluconate
KG: 2-ketogluconate
mms: maltose metabolism stimulation
ORF:
open reading frames
SDS-PAGE: sodium dodecyl sulphate-spolyacrylamide gel electrophoresis
|
We cloned
2-keto-3-deoxy-gluconate kinase (KDGK), which catalyzes the phosphorylation of
2-keto-3-deoxygluconate (KDG) to 2-keto-3-deoxy-6-phophogluconate (KDPG) from Serratia
marcescens KCTC 2172. The nucleotide sequence revealed a single open
reading frame containing 1,208 bp and encoding for 309 amino acids, with a
molecular weight of 33,993 Da. The enzyme was purified via GST affinity
chromatography. The putative KdgT binding site was detected upstream of the initial
codon. The KDG kinase utilized 2-ketogluconate (KG) and KDG as substrates. The
optimal temperature and pH for KDGK activity were 50ºC and 8.0, respectively.
|
