Favored isolation and rapid identification of the astaxanthin-producing yeastXanthophyllomyces dendrorhous(Phaffia rhodozyma) from environmental samples (original) (raw)
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Journal of Industrial Microbiology & Biotechnology, 2007
A novel population of the biotechnologically important yeast Xanthophyllomyces dendrorhous, the sexual stage of PhaYa rhodozyma, has been recently isolated for the Wrst time in the southern Hemisphere (Patagonia, Argentina). The aim of the present work was to phenotypically and genotypically characterize two representative strains of this new population, and assess such strains as a potential biotechnological source of astaxanthin, fatty acids and extracellular enzymes. Minor variations were found in physiological tests. PCR Wngerprinting studies (MSP-PCR) showed the main diVerences between X. dendrorhous Patagonian and Type strains. Patagonian strains accumulated a xanthophyll-like pigment, which was identiWed as astaxanthin. These strains showed low fatty acids content (mainly polyunsaturated fatty acids) and, of a total of six extracellular enzymes tested, only produced amylase. Genetic diVerences between Patagonian and collection X. dendrorhous strains could be explained by geographic isolation and habitat speciWcity.
2004
Astaxanthin (3,3 ABSTRACT ʹ-dihydroxy-β,β-carotene-4,4ʹ-dione)-accumulating yeasts are of great biotechnological interest. Random amplifi ed polymorphic DNA (RAPD) analysis involving 5 primers and 13 astaxanthin-producing yeast strains was performed. Cluster analysis based on RAPD markers differentiated isolates of Xanthopyllomyces dendrorhous (self-sporulating) and Phaffi a rhodozyma (asexual) at an intraspecifi c level. Strains considered to be derived from the same isolate, but which had had different strain histories, revealed signifi cant differences in their RAPD patterns. The applicability of RAPD analysis for the species-level differentiation of these yeasts is discussed.
Acta scientiarum polonorum. Technologia alimentaria
Astaxanthin is the most important and expensive carotenoid pigment used in aquaculture. Its commercial attractiveness is also related with its antioxidant potential. Xanthophyllomyces dendrorhous yeast is considered to be promising for commercial production of astaxanthin. The aim of this study was to investigate the possibility of the growth and astaxanthin production by X. dendrorhous strains 011 media containing xylose under different illumination. A', dendrorhous DSM 5626 and its mutants: 10BE and 26UV were used in this study. The cultures were carried out 011 hydrolysed rye stillage (HS) and YM medium with xylose (YM-K). Cell concentration, total carotenoid and astaxanthin yields were assessed in 5-day cultures. The effect of illumination in the range of 0-5.000 lx 011 growth and on astaxanthin production of yeasts in cultures run 011 YM-K medium was also examined. For the tested yeast strains better growth parameters and astaxanthin yields were obtained on the YM-K medium....
Xanthophyllomyces dendrorhous for the industrial production of astaxanthin
Applied Microbiology and Biotechnology, 2010
Astaxanthin is a red xanthophyll (oxygenated carotenoid) with large importance in the aquaculture, pharmaceutical, and food industries. The green alga Haematococcus pluvialis and the heterobasidiomycetous yeast Xanthophyllomyces dendrorhous are currently known as the main microorganisms useful for astaxanthin production at the industrial scale. The improvement of astaxanthin titer by microbial fermentation is a requirement to be competitive with the synthetic manufacture by chemical procedures, which at present is the major source in the market. In this review, we show how the isolation of new strains of X. dendrorhous from the environment, the selection of mutants by the classical methods of random mutation and screening, and the rational metabolic engineering, have provided improved strains with higher astaxanthin productivity. To reduce production costs and enhance competitiveness from an industrial point of view, low-cost raw materials from industrial and agricultural origin have been adopted to get the maximal astaxanthin productivity. Finally, fermentation parameters have been studied in depth, both at flask and fermenter scales, to get maximal astaxanthin titers of 4.7 mg/g dry cell matter (420 mg/l) when X. dendrorhous was fermented under continuous white light. The industrial scale-up of this biotechnological process will provide a cost-effective method, alternative to synthetic astaxanthin, for the commercial exploitation of the expensive astaxanthin (about $2,500 per kilogram of pure astaxanthin).
Polish journal of microbiology / Polskie Towarzystwo Mikrobiologów = The Polish Society of Microbiologists, 2013
Astaxanthin is a xanthophill pigment with commercial application in the aquaculture, pharmaceutical, food and cosmetic industries. The red yeast Xanthophyllomyces dendrorhous is one of the most promising microorganisms for its industrial production. However, astaxanthin content in wild yeast strains is low. Pigment production by X. dendrorhous can be improved by mutagenesis. The aim of the study was to assess the efficiency of four mutagens: UV radiation, benomyl, ethyl methanesulfonate and ethidium bromide in generating asthaxanthin-hyperproducing strains of the yeast Xanthophyllomyces dendrorhous DSM 5626. Mutations with benomyl, ethidium bromide and UV radiation generated a group of hyperpigmented mutants exhibiting increases up to 100% in astaxanthin content. Ethyl methanesulfonate turned out to be useless in this respect.
World Journal of Microbiology and Biotechnology, 2015
Antarctic microorganisms have developed different strategies to live in their environments, including modifications to their membrane components to regulate fluidity and the production of photoprotective metabolites such as carotenoids. Three yeast colonies (ANCH01, ANCH06 and ANCH08) were isolated from soil samples collected at King George Island, which according to their rDNA sequence analyses, were determined to be Xanthophyllomyces dendrorhous. This yeast is of biotechnological interest, because it can synthesize astaxanthin as its main carotenoid, which is a powerful antioxidant pigment used in aquaculture. Then, the aim of this work was to characterize the ANCH isolates at their molecular and phenotypic level. The isolates did not display any differences in their rDNA and COX1 gene nucleotide sequences. However, ANCH01 produces approximately sixfold more astaxanthin than other wild type strains. Moreover, even though ANCH06 and ANCH08 produce astaxanthin, their main carotenoid was b-carotene. In contrast to other X. dendrorhous strains, the ANCH isolates did not produce mycosporines. Finally, the ANCH isolates had a higher proportion of polyunsaturated fatty acids than other wild type strains. In conclusion, the reported X. dendrorhous isolates are phenotypically different from other wild type strains, including characteristics that could make them more resistant and better able to inhabit their original habitat, which may also have biotechnological potential.
Journal of Basic Microbiology, 2009
The yeast Xanthophyllomyces dendrorhous is biotechnologically important due to its ability to produce the pigment astaxanthin, but is poorly understood at the genetic level. This is mainly because its preservation is difficult and many of the mutants obtained are unstable. The objectives of the present work were (i) the mutagenesis X. dendrorhous and, (ii) isolation of mutants with auxotrophic markers suitable for genetic studies of the carotenogenesis pathway and sexual cycle. Additionally, two kinds of preservation methods at the laboratory level were tested for the storage of strains. A collection of X. dendrorhous mutants affected in the production of carotenoid pigments or development of sexual structures and auxotrophic requirements were isolated by treatment with N-methyl-N′-nitro-N-nitrosoguanidine and the antibiotic nystatin. From a detailed analysis about the requirements of auxotrophic mutants the ARG7, ARG3 and PRO3 loci can be defined in this yeast. Among the methods assayed for the long-term preservation of X. dendrorhous strains, the dehydrated gelatin drop method showed the highest recovery of viable yeast after storage for 65 months. No changes in auxotrophic properties and in macro or micro morphology were observed after applying the latter method. (© 2009 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim)
Simple method for the isolation of astaxanthin from the basidiomycetous yeast Phaffia rhodozyma
Applied and environmental microbiology, 1978
A method is described for the quantitative and, possibly, large-scale extraction of astaxanthin from the yeast Phaffia rhodozyma. The method utilizes extracellular enzymes produced by the bacterium Bacillus circulans WL-12, which partially digests the yeast cell wall and renders the carotenoid pigments extractable by acetone or ethanol. Complete recovery of astaxanthin from heat-killed P. rhodozyma cells was obtained after growing B. circulans WL-12 on these yeast cells for 26 h and then extracting the yeast-bacterium mixture with acetone. A bacteria-free lytic system, which gave quantitative extraction of astaxanthin from P. rhodozyma, was obtained by concentrating the culture broth from the growth of B. circulans WL-12 on P. rhodozyma cells. Hydrolytic enzyme activities detected in this concentrate included beta-(1 leads to 3)-glucanase, beta-(1 leads to 6)-glucanase, alpha-(1 leads to 3)-glucanase, xylanase, and chitinase. The lytic system was found to work most efficiently at pH...
Stimulation of astaxanthin formation in the yeast by the fungus
FEMS Yeast Research, 2004
A fungal contaminant on an agar plate containing colonies of Xanthophyllomyces dendrorhous markedly increased carotenoid production by yeast colonies near to the fungal growth. Spent-culture filtrate from growth of the fungus in yeast^malt medium also stimulated carotenoid production by X. dendrorhous. Four X. dendrorhous strains including the wild-type UCD 67-385 (ATCC 24230), AF-1 (albino mutant, ATCC 96816), Yan-1 (L-carotene mutant, ATCC 96815) and CAX (astaxanthin overproducer mutant) exposed to fungal concentrate extract enhanced astaxanthin up to approximately 40% per unit dry cell weight in the wild-type strain and in CAX. Interestingly, the fungal extract restored astaxanthin biosynthesis in non-astaxanthin-producing mutants previously isolated in our laboratory, including the albino and the L-carotene mutant. The fungus was identified as Epicoccum nigrum by morphology of sporulating cultures, and the identity confirmed by genetic characterization including rDNA sequencing analysis of the large-subunit (LSU), the internal transcribed spacer, and the D1/D2 region of the LSU. These E. nigrum rDNA sequences were deposited in GenBank under accesssion numbers AF338443, AY093413 and AY093414. Systematic rDNA homology alignments were performed to identify fungi related to E. nigrum. Stimulation of carotenogenesis by E. nigrum and potentially other fungi could provide a novel method to enhance astaxanthin formation in industrial fermentations of X. dendrorhous and Phaffia rhodozyma.