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The effect of gamma irradiation on astaxanthin synthetase encoding gene in two mutant strains of Phaffia rhodozyma.

ABSTRACT: BACKGROUND AND OBJECTIVES:Astaxanthin, an orange-red carotenoid pigment, acts as a protective agent against oxidative damage to cells in vivo. The astaxanthin synthetase gene (crtS) size consists of 3995 bp. This gene has been suggested to catalyse ?-carotene to astaxanthin in Phaffia rhodozyma. The aim of this research was to find any possible changes in this gene in two mutant strains, Gam1 and Gam2 (with high astaxanthin pigment production), previously created by gamma irradiation. MATERIALS AND METHODS:The astaxanthin synthetase gene sequence of Phaffia rhodozyma in the NCBI Gene bank was used to design primer. In Gam1, this gene was amplified using primers Asta F1, Asta R2, Asta F3, Asta R4. In Gam2, primers asta F1, asta R4 were used to amplify the gene. The amplified fragments were 8 sequenced using primers Asta F1, Asta R1, Asta F2, Asta R2, Asta F3, Asta R3 and Asta F4, Asta R4. Astaxanthin synthetase gene from two mutant strains, Gam1 and Gam2 were amplified using PCR. The amplified products were sequenced and aligned using the ClustalW software. CONCLUSION:The comparison of this gene showed 98% and 99% similarities between the reference sequence and Gam1 and Gam2 mutant strains, respectively, whereas the comparison of this gene in Gam1 and Gam2 mutant strains showed 97% similarity. However, the deduced proteins showed 78% and 83% between the reference protein obtained from the wild type and Gam1 and Gam2, respectively. This similarity was 75% between the mutant strains.

SUBMITTER: Najafi N

PROVIDER: S-EPMC3895570 | biostudies-literature | 2013 Sep

REPOSITORIES: biostudies-literature

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The effect of gamma irradiation on astaxanthin synthetase encoding gene in two mutant strains of Phaffia rhodozyma.

Najafi Naeimeh N Hosseini Ramin R Ahmadi Ali-Reza AR

Iranian journal of microbiology 20130901 3

<h4>Background and objectives</h4>Astaxanthin, an orange-red carotenoid pigment, acts as a protective agent against oxidative damage to cells in vivo. The astaxanthin synthetase gene (crtS) size consists of 3995 bp. This gene has been suggested to catalyse β-carotene to astaxanthin in Phaffia rhodozyma. The aim of this research was to find any possible changes in this gene in two mutant strains, Gam1 and Gam2 (with high astaxanthin pigment production), previously created by gamma irradiation.<h4 ...[more]

PMID: 24475339

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Project description:Phaffia rhodozyma represents an excellent microbial resource for astaxanthin production. However, the yeast's low astaxanthin productivity poses challenges in scaling up industrial production. Although P. rhodozyma originates from plant material, and phytohormones have demonstrated their effectiveness in stimulating microbial production, there has been limited research on the effects and mechanisms of phytohormones on astaxanthin biosynthesis in P. rhodozyma. In this study, the addition of exogenous salicylic acid (SA) at a concentration as low as 0.5 mg/L significantly enhanced biomass, astaxanthin content, and yield by 20.8%, 95.8% and 135.3% in P. rhodozyma, respectively. Moreover, transcriptomic analysis showed that SA had discernible impact on the gene expression profile of P. rhodozyma cells. Differentially expressed genes (DEGs) in P. rhodozyma cells between the SA-treated and SA-free groups were identified. These genes played crucial roles in various aspects of astaxanthin and its competitive metabolites synthesis, material supply, biomolecule metabolite and transportation, anti-stress response, and global signal transductions. This study proposes a regulatory mechanism for astaxanthin synthesis induced by SA, encompassing the perception and transduction of SA signal, transcription factor-mediated gene expression regulation, and cellular stress responses to SA. Notably, the polyamine transporter gene (PT), identified as an upregulated DEG, was overexpressed in P. rhodozyma to obtain the transformant Prh-PT-006. The biomass, astaxanthin content and yield in this engineered strain could reach 6.6 g/L, 0.35 mg/g DCW and 2.3 mg/L, 24.5%, 143.1% and 199.0% higher than the wild strain at the SA-free condition, respectively. These findings provide valuable insights into potential targets for genetic engineering aimed at achieving high astaxanthin yields, and such advancements hold promise for expediting the industrialization of microbial astaxanthin production.

Project description:The present study aimed to investigate the effect of dietary astaxanthin (Ast) from Phaffia rhodozyma on growth performance, survival, carotenoid content, the activity of antioxidant and immune-related enzymes, intestinal microbiota comparison, and disease resistance against Vibrio parahaemolyticus in Penaeus monodon. Juveniles (average weight 3.15 ± 0.12 g) were fed with six experimental diets supplemented with 0 (Control), 20.5, 41, 61.5, 82, and 102.5 mg/kg of Ast (defined as diet A-D) in triplicate for 56 days. The results indicated that shrimp fed with Ast supplementation significantly (p < 0.05) improved growth performance compared with the control. Furthermore, significantly (p < 0.05) increased survival and decreased feed conversion ratio (FCR) demonstrated the beneficial effects of dietary Ast on enhancing nutrient utilization and ultimately improving the growth and survival of shrimp. Furthermore, shrimp fed with Ast including diet developed a deeper red color than the control, consistent with the significantly (p < 0.05) increased Ast deposition in the shrimp shell. Hemolymph-immunological parameters [aspartate aminotransferase (AST), alanine aminotransferase (ALT), and alkaline phosphatase (AKP)] and hepatopancreatic antioxidant status [total antioxidant capacity (T-AOC), malondialdehyde (MDA), catalase (CAT), and superoxide dismutase (SOD)] were significantly (p < 0.05) affected by dietary Ast supplementation. Dietary increasing Ast levels significantly (p < 0.05) increased shrimp resistance performance to V. parahaemolyticus according to the LT50 results in the current study, which may be caused by increased total carotenoid contents in shrimp tissues from all the Ast-supplemented treatments. Conversely, intestinal microbiota biodiversity and richness were not affected by dietary Ast. The best performances of growth, antioxidant status, immunological response, and carotenoid deposition were observed in diets E and F among all the Ast-supplemented treatments. Overall, all the data suggested that dietary P. rhodozyma Ast played a critical role in improving growth performance, achieving the desired coloration, increasing carotenoid content, and keeping better health status of shrimp. Based on these positive performances, P. rhodozyma Ast could gain the trust of the consumers as a natural source and provide a potential alternative for synthetic Ast using in the Penaeus monodon culture industry.

Dataset Information

The effect of gamma irradiation on astaxanthin synthetase encoding gene in two mutant strains of Phaffia rhodozyma.

Publications

The effect of gamma irradiation on astaxanthin synthetase encoding gene in two mutant strains of Phaffia rhodozyma.

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