Cloning and Expression of Phytoene Desaturase Gene from Undaria pinnatifida Suringar
Tengteng Guan, Tingting Zhang, Yi Ma, Yinjie Li, Guicai Du* and Ronggui Li** Author for corresponding; e-mail address: duguicai@qdu.edu.cn; lrg@qdu.edu.cn
Volume: Vol.47 No.6 (November 2020)
Research Article
DOI:
Received: 2 July 2019, Revised: -, Accepted: 18 June 2020, Published: -
Citation: Guan T., Zhang T., Ma Y., Li Y., Du G. and Li R., Cloning and Expression of Phytoene Desaturase Gene from Undaria pinnatifida Suringar, Chiang Mai Journal of Science, 2020; 47(6): 1118-1129.
Abstract
The phytoene desaturase is a key enzyme involved in the carotenoids biosynthesis pathway, which catalyzes the conversion of ζ-carotene from phytoene. Undaria pinnatifida Suringar is an edible economic brown algae that produces high concentrations of fucoxanthin. In this study, the PDS gene (UpPDS) was isolated and characterized from U. pinnatifida, and the full-length cDNA sequence was 1707 bp in length and encoded 568 amino acid residues. Furthermore, the putative protein sequence showed a high homology to the PDS from other known plants. Phylogenetic analysis showed that the UpPDS was more related to that of brown algae compared to other species. By heterologous complementation and high-performance liquid chromatography analysis, desaturation activity of recombinant UpPDS was confirmed by production of ζ-carotene in engineering E. coli co-transformed with crtE, crtB and UpPDS. The L9(3 4) orthogonal test was used to optimize ζ-carotene fermentation conditions of the engineering bacteria. The results showed that temperature had the most significant effect on ζ-carotene production, the maximal yield of ζ-carotene could reach 5.0mg/L under the optimal conditions.