Ethanol as a carotenoid production stimulator in Dunaliella salina CCAP 19/18.

Dunaliella salina is a rich source of carotenoids. Carotenoid production is induced under specific conditions, i.e., high light intensity, high salt concentration, nutrient limitation, and suboptimal temperatures in this microalga. The control of environmental factors is vital for high productivity of carotenoids. In this paper, the effect of different ethanol concentrations in combination with nitrogen deficiency was investigated to induce carotenoid production in D. salina CCAP 19/18. Also, some biochemical and molecular parameters were investigated in response to ethanol in the cells. It was shown that ethanol at 0.5% concentration increased cell number but, at 5% concentration, reduced cell viability compared to the control. The highest carotenoid production was achieved at 3% ethanol concentration, which was 1.46 fold higher than the nitrogen deficiency condition. Investigation of the 3 carotenoid biosynthesis genes revealed that their expression levels increased at 3% ethanol concentration, and the phytoene synthase gene was the most upregulated one. Lipid peroxidation increased at both 3% and 5% ethanol concentrations. At 3% concentration, the activity of catalase and superoxide dismutase increased, but no significant changes were seen at 5% ethanol concentration. Peroxidase activity reduced at both 3% and 5% concentrations. Moreover, proline and reducing sugar content increased at 3% concentration while decreased at 5% ethanol concertation. The results showed that at 3% ethanol concentration, higher carotenoid productivity was associated with an increase in other intracellular responses (molecular and biochemical). Ethanol as a controllable element may be beneficial to increase carotenoid production even under inappropriate environmental conditions in D. salina.

Genetic and in silico analysis of plantaricin EFI locus in indigenous isolates of lactobacillus plantarum.

Genetic investigation and in silico analysis of plantaricin EFI (plnEFI) locus was performed in three indigenous isolates of Lactobacillus plantarum EL3, L28 and BL1. Amplification with plnEFI specific primers and production of ~ 10 KDa size protein suggested the existence of class II bacteriocins. The analysis demonstrated that the studied fragment included structural bacteriocin, immunity, partial transporter and potential regulatory encoding regions. Based on the results, there was one DNA polymorphic site in plnE as well as plnF of the studied sequences. One nucleotide substitution in plnE of BL1 isolate lead to replacement of Glycin with Valine. These two are of non-polar type which did not affect instability index of plnE protein. The only nucleotide variation in plnF of EL3 isolate did not change the amino acid sequence since the modified nucleotide constituted alternative codon of the original amino acid. The highest DNA polymorphism occurred in the region with immunity function which in BL1 resulted in the conversion of start codon to amino acid codon. In the partial transporter sequence, one variable nucleotide site caused amino acid replacement in all the isolates which elevated stability of N-terminal domain in the transporter protein compared to nominated reference isolate L. plantarum C11. The region with possible regulatory function was identical in all three isolates. © 2018 American Institute of Chemical Engineers Biotechnol Progress, 35: e2773, 2019.

Introduction of a novel 18S rDNA gene arrangement along with distinct ITS region in the saline water microalga Dunaliella.

Comparison of 18S rDNA gene sequences is a very promising method for identification and classification of living organisms. Molecular identification and discrimination of different Dunaliella species were carried out based on the size of 18S rDNA gene and, number and position of introns in the gene. Three types of 18S rDNA structure have already been reported: the gene with a size of ~1770 bp lacking any intron, with a size of ~2170 bp consisting one intron near 5' terminus, and with a size of ~2570 bp harbouring two introns near 5' and 3' termini. Hereby, we report a new 18S rDNA gene arrangement in terms of intron localization and nucleotide sequence in a Dunaliella isolated from Iranian salt lakes (ABRIINW-M1/2). PCR amplification with genus-specific primers resulted in production of a ~2170 bp DNA band, which is similar to that of D. salina 18S rDNA gene containing only one intron near 5' terminus. Whilst, sequence composition of the gene revealed the lack of any intron near 5' terminus in our isolate. Furthermore, another alteration was observed due to the presence of a 440 bp DNA fragment near 3' terminus. Accordingly, 18S rDNA gene of the isolate is clearly different from those of D. salina and any other Dunaliella species reported so far. Moreover, analysis of ITS region sequence showed the diversity of this region compared to the previously reported species. 18S rDNA and ITS sequences of our isolate were submitted with accesion numbers of EU678868 and EU927373 in NCBI database, respectively. The optimum growth rate of this isolate occured at the salinity level of 1 M NaCl. The maximum carotenoid content under stress condition of intense light (400 mumol photon m-2 s-1), high salinity (4 M NaCl) and deficiency of nitrate and phosphate nutritions reached to 240 ng/cell after 15 days.