Cloning and transcription analysis of the nitrate reductase gene from Haematococcus pluvialis

Biotechnol Lett. 2017 Apr;39(4):589-597. doi: 10.1007/s10529-016-2283-0. Epub 2017 Jan 4.

Abstract

Objectives: To optimize the cultivation media for the growth rate of Haematococcus pluvialis and to study the transcription regulation of the algal nitrate reductase (NR), a key enzyme for nitrogen metabolism.

Results: The NR gene from H. pluvialis hd7 consists of 5636 nucleotides, including 14 introns. The cDNA ORF is 2718 bp, encoding a 905 aa protein with three conserved domains. The NR amino acids of H. pluvialis hd7 are hydrophilic and have similarity of 72% compared to that of Dunaliella. NR transcription increased with an increase of nitrate concentration from 0.4 to 1 g/l. A deficiency of nitrogen increased NR transcription significantly. The transcription level of NR increased at phosphorus concentrations from 0.08 to 0.2 g/l, with a maximum at 0.08 g/l. The optimum parameters of medium component for transcription of NR and growth of H. pluvialis were 0.3 g NaNO3/l, 0.045 g KH2PO4/l and 1.08 g sodium acetate/l.

Conclusions: This study provides a better understanding of nitrate regulation in H. pluvialis.

Keywords: Astaxanthin; Haematococcus pluvialis; Nitrate reductase; Nitrogen metabolism.

MeSH terms

  • Acetic Acid / metabolism
  • Algal Proteins / genetics*
  • Amino Acid Sequence
  • Cell Culture Techniques
  • Chlorophyta / enzymology*
  • Chlorophyta / genetics
  • DNA, Algal / genetics
  • Gene Expression*
  • Nitrate Reductase / genetics*
  • Nitrates / metabolism*
  • Nitrogen / metabolism
  • Phosphorus / metabolism
  • Transcription, Genetic

Substances

  • Algal Proteins
  • DNA, Algal
  • Nitrates
  • Phosphorus
  • Nitrate Reductase
  • Nitrogen
  • Acetic Acid