Identification, characterization and quantitative analysis of NAD-malate dehydrogenase from the marine rhodophyte Pyropia haitanensis

Bao Yu Zhang; Zhao Jun Hou; Guang Ce Wang; Guang Peng

doi:10.1515/bot-2014-0052

Article

Identification, characterization and quantitative analysis of NAD-malate dehydrogenase from the marine rhodophyte Pyropia haitanensis

Bao Yu Zhang
Bao Yu Zhang is an Associate Professor at the Institute of Oceanology, Chinese Academy of Sciences. Her major research interest is the molecular biology of marine algae, focusing especially on inorganic carbon fixation and lipid synthesis pathways in algae.
, Zhao Jun Hou
Zhao Jun Hou is an instructor in the College of Computer Science and Information Engineering, Tianjin University of Science and Technology. He obtained his Master’s degree in the Tianjin University of Science and Technology.
, Guang Ce Wang
Guang Ce Wang is a Professor at the Institute of Oceanology, Chinese Academy of Sciences. His research interest is marine algal physiology and developmental regulation.
and Guang Peng
Guang Peng is a technician in the Institute of Oceanology, Chinese Academy of Sciences. Mr. Peng has many years of experience in marine algal cultivation and has established an efficient indoor cultivation system.

Published/Copyright: June 22, 2015

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From the journal Botanica Marina Volume 58 Issue 4

Abstract

Malate dehydrogenase (MDH) is an enzyme that catalyzes the interconversion of malate and oxaloacetate substrates and is widely distributed from prokaryotes to eukaryotes. It plays crucial roles in many important metabolic pathways and includes different isoforms based on coenzyme specificity and cellular localization. To study MDH in rhodophytes, we obtained a full-length cDNA clone (here designated PhMDH) encoding an NAD-malate dehydrogenase in the marine red alga Pyropia haitanensis. The nucleotide sequence of PhMDH was 1521 bp, including an open reading frame (ORF) of 984 bp. The amino acid sequence showed 73% identity with other MDHs in proteobacteria. Two MDH-like domains were detected in the 5–145 and 156–320 regions. Real-time fluorescent quantitative PCR (qPCR) was used to examine mRNA expression levels during the gametophyte and sporophyte phases. The transcription of PhMDH in the gametophyte was barely detectable, whereas PhMDH in the sporophyte showed a much higher expression level. The activity of PhMDH in the filamentous sporophyte was approximately double that of the leafy gametophyte. Considering these results, we suggest that PhMDH may be localized in the cytosol and play a role in carbon fixation in the sporophyte stage.

Keywords: activity; NAD-malate dehydrogenase; qPCR

Corresponding author: Guang Ce Wang, Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, China, e-mail: gcwang@qdio.ac.cn

aThese authors contributed equally to this work.

About the authors

Bao Yu Zhang

Bao Yu Zhang is an Associate Professor at the Institute of Oceanology, Chinese Academy of Sciences. Her major research interest is the molecular biology of marine algae, focusing especially on inorganic carbon fixation and lipid synthesis pathways in algae.

Zhao Jun Hou

Zhao Jun Hou is an instructor in the College of Computer Science and Information Engineering, Tianjin University of Science and Technology. He obtained his Master’s degree in the Tianjin University of Science and Technology.

Guang Ce Wang

Guang Ce Wang is a Professor at the Institute of Oceanology, Chinese Academy of Sciences. His research interest is marine algal physiology and developmental regulation.

Guang Peng

Guang Peng is a technician in the Institute of Oceanology, Chinese Academy of Sciences. Mr. Peng has many years of experience in marine algal cultivation and has established an efficient indoor cultivation system.

Acknowledgments

This work was supported by the National High Technology Research and Development Program of China (863 Program) (No. 2012AA100806), National Natural Science Foundation of China (41176134, 41306151), Marine Public Project (201105023-7) and 973 Project (2011CB411908).

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Received: 2014-8-27

Accepted: 2015-5-20

Published Online: 2015-6-22

Published in Print: 2015-8-1

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Keywords for this article

activity; NAD-malate dehydrogenase; qPCR