Elsevier

Process Biochemistry

Volume 63, December 2017, Pages 16-25
Process Biochemistry

Astaxanthin from Jerusalem artichoke: Production by fed-batch fermentation using Phaffia rhodozyma and application in cosmetics

https://doi.org/10.1016/j.procbio.2017.08.013Get rights and content

Highlights

  • Jerusalem artichoke is suitable for astaxanthin production by P. rhodozyma Y119.

  • Substrate feedback feeding is most effective for carotenoid production.

  • Astaxanthin essence was produced from extracted astaxanthin.

  • The essence is stable at 4 °C with a 70-week half-life.

Abstract

Jerusalem artichoke extract or powder was used for astaxanthin production using Phaffia rhodozyma without acidic or enzymatic inulin hydrolysis. The culture medium containing Jerusalem artichoke as carbon source was optimized, and feeding strategies, including constant, exponential, pH-stat, and substrate feedback fed-batch fermentations, were also compared for enhancing the cell biomass and astaxanthin synthesis by P. rhodozyma. Substrate-feedback fed-batch fermentation resulted in the highest dry cell weight of 83.60 g/L, with a carotenoid concentration and yield of 982.50 mg/L and 13.30 mg/g, respectively, under optimized medium components using Jerusalem artichoke extract as carbon source in a 3-L stirred-tank bioreactor. Moreover, 482.50 mg/L of carotenoids and 253.10 mg/L of astaxanthin were obtained by continuous feeding of Jerusalem artichoke powder, which was used as carbon source. Astaxanthin essence with high DPPH-scavenging activity was obtained from the extracted astaxanthin, and the DPPH free radical scavenging rate of 40 ppm astaxanthin essence reached 76.29%. When stored at 4 °C, astaxanthin essence showed the highest stability, with a minimum k value of 0.0099 week−1 and maximum half-life (t1/2) value of 70 weeks.

Introduction

Astaxanthin (3,3′-dihydroxy-β,β’-carotene-4,4′-dione) has strong antioxidant, anti-aging, anti-inflammatory, sun proofing, and immune system-boosting functions, suggesting its great commercial potential in pharmaceutical, food, and feed industries [1], [2]. In recent years, astaxanthin from the red yeast Phaffia rhodozyma or the microalga Haematococcus pluvialis has been widely investigated as a potential natural pigment source [3]. Despite lower astaxanthin production from yeasts than algae, the former has the advantages of higher growth rate, easier cultivation conditions, and shorter production time at industrial scale [4]. However, commercial production of astaxanthin from P. rhodozyma is hampered by relatively low productivity and high costs due to strain instability and the tendency of mutants to produce less astaxanthin [5]. Therefore, it is crucial to improve astaxanthin productivity and decrease the production costs.

In recent years, a variety of agro-industrial products or by-products have been explored as substrates for microbial astaxanthin production, e.g., enzymatic eucalyptus wood hydrolysate [6], mustard waste precipitated hydrolysate [7], corn starch hydrolysate [8], yucca date juice [9], [10], sugarcane juice [11], and mussel processing wastewater [12]. However, to the best of our knowledge, Jerusalem artichokes have not yet been investigated for the cultivation of P. rhodozyma despite their high content of nutritive compounds that can be used as human and animal food. As an inexpensive alternative feedstock, the tubers of Jerusalem artichoke are rich in inulin, which can even reach to more than 50% in the dried tubers [13], [14]. As a reserve carbohydrate, inulin is a linear polymer of D-fructose containing a terminal D-glucose, which can be hydrolyzed to fructose, sucrose, glucose and polymers of fructo-oligosaccharides by inulinase. In addition, unlike grain crops, Jerusalem artichoke can grow well in barren land, with the advantages of drought and saline tolerance; moreover, it does not compete with grain crops for arable land [15]. Despite its various advantages, the use of Jerusalem artichoke in the production of high value-added products has been hindered by the high degree of polymerization. Currently, after pretreatment by acid or inulinase hydrolysis, Jerusalem artichoke can be used as a substrate to produce ethanol, single-cell oil, and single-cell protein [13], [16], [17], [18]. However, there are no reports on whether astaxanthin can be produced from Jerusalem artichoke and whether Jerusalem artichoke can be fermented by P. rhodozyma without acid or enzymatic hydrolysis.

In addition, a desirable medium and a suitable feeding strategy are also essential for massive accumulation of astaxanthin. Response surface methodology in combination with central composite design is widely used for medium optimization [19]. Optimum low-cost substrates for astaxanthin biosynthesis could be selected and designed by central composite design and optimized by response surface methodology. Furthermore, fed-batch fermentations are commonly used because of the substantial effect of varying substrate concentrations on cell biomass and metabolite production in many fermentations [20]. The feeding amount and feeding time also play a very important role in maximizing the production of desired products [21]. Despite extensive studies concerning the optimization of substrate feeding for fed-batch fermentation process, to the best of our knowledge, there is no report available on the substrate feedback feeding strategy using Jerusalem artichoke as substrate for enhanced microbial astaxanthin production.

Thus far, astaxanthin has been mainly used as a feed additive in aquaculture for appropriate pigmentation, growth, and reproduction of trout [22]. However, more and more health-promoting properties of astaxanthin have been revealed. This opens up new possibilities for its potential application as a nutraceutical or as a constitutional ingredient in cosmetics [23], [24]. Despite reports about a variety of astaxanthin healthcare products, little information is available about the direct application of astaxanthin and its free radical-scavenging capacity in cosmetics.

In this study, Jerusalem artichoke was used for astaxanthin production by P. rhodozyma without acid or enzymatic hydrolysis. The culture medium with Jerusalem artichoke extract or powder as carbon source was optimized, and optimum feeding strategies for enhancing the cell biomass and astaxanthin synthesis by P. rhodozyma were also explored. Moreover, an astaxanthin essence with high DPPH-scavenging activity was obtained from the extracted astaxanthin.

Section snippets

Chemicals and reagents

Acetonitrile, methanol (MeOH), and acetone (all HPLC grade) were obtained from Merck Chemical Co. (Darmstadt, Germany). Astaxanthin standard was purchased from Sigma-Aldrich Chemical Co. (St. Louis, MO, USA). All other chemical reagents were of analytical grade and procured from standard companies.

Raw material and its pretreatment

The Jerusalem artichoke crude material was purchased from a local market. The commercially purchased fresh Jerusalem artichoke crude material contained about (w/w) 58.07 ± 0.69% inulin; 1.54 ± 0.03%

Optimization of medium with Jerusalem artichoke extract and powder as carbon sources

To determine the optimal concentration of Jerusalem artichoke extract for carotenoid accumulation and biomass, a series of experiments were performed in flasks. P. rhodozyma Y119 could directly metabolize inulin from Jerusalem artichoke for the production of carotenoids without acidic or enzymatic hydrolysis. The growth and carotenoid production in Jerusalem artichoke extract medium were comparable to those in the sucrose medium (data not shown). The optimal medium for carotenoid production

Conclusions

Jerusalem artichoke could be efficiently converted to astaxanthin by P. rhodozyma Y119 without acidic or enzymatic hydrolysis. Substrate feedback feeding strategy was the most effective process for biomass and carotenoid production, leading to a maximum carotenoid production of 982.50 mg/L and DCW of 83.60 g/L. Moreover, the extracted astaxanthin was developed into an essence with a high DPPH-scavenging activity. The present study indicates that Jerusalem artichoke is a promising raw material for

Acknowledgements

This research was supported by Guangzhou Science and Technology Program [grant No.2014Y2-00515].

References (52)

  • H. Sun et al.

    Enhancement of cell biomass and cell activity of astaxanthin-rich Haematococcus pluvialis

    Bioresour. Technol.

    (2015)
  • L.M. Monks et al.

    Use of chemical, enzymatic and ultrasound-assisted methods for cell disruption to obtain carotenoids

    Biocatalysis Agricultural Biotechnol.

    (2013)
  • N. Anarjan et al.

    Colloidal astaxanthin: Preparation, characterisation and bioavailability evaluation

    Food Chem.

    (2012)
  • Y. Sun et al.

    Stability of all-trans-beta-carotene under ultrasound treatment in a model system: effects of different factors, kinetics and newly formed compounds

    Ultrason. Sonochem.

    (2010)
  • T. Zhang et al.

    Bioethanol production from hydrolysates of inulin and the tuber meal of Jerusalem artichoke by Saccharomyces sp. W0

    Bioresour. Technol.

    (2010)
  • Y.S. Liu et al.

    Characterization of oxygen transfer conditions and their effects on Phaffia rhodozyma growth and carotenoid production in shake-flask cultures

    Biochem. Eng. J.

    (2006)
  • J. Takeuchi et al.

    Preparation of dried chips from Jerusalem artichoke (Helianthus tuberosus) tubers and analysis of their functional properties

    Food Chem.

    (2011)
  • A.J. Melendezmartinez et al.

    Relationship between the colour and the chemical structure of carotenoid pigments

    Food Chem.

    (2007)
  • F. Ahmed et al.

    Effect of drying storage temperature and air exposure on astaxanthin stability from Haematococcus pluvialis

    Food Res. Int.

    (2015)
  • D.K. Kim et al.

    Transcriptomic analysis of Haematococcus lacustris during astaxanthin accumulation under high irradiance and nutrient starvation

    Biotechnol. Bioproc. E

    (2011)
  • L.C. Mata-Gómez et al.

    Biotechnological production of carotenoids by yeasts: an overview

    Microb. Cell Fact.

    (2014)
  • I. Schmidt et al.

    Biotechnological production of astaxanthin with Phaffia rhodozyma/Xanthophyllomyces dendrorhous

    Appl. Microbiol. Biotechnol.

    (2011)
  • J. Montanti et al.

    Production of Astaxanthin from Cellulosic Biomass Sugars by Mutants of the Yeast Phaffia rhodozyma

    Appl. Biochem. Biotechnol.

    (2011)
  • J. Tinoi et al.

    Utilization of mustard waste isolates for improved production of astaxanthin by Xanthophyllomyces dendrorhous

    J. Ind. Microbiol. Biotechnol.

    (2006)
  • S. Siva Kesava et al.

    An industrial medium for improved production of carotenoids from a mutant strain of Phaffia rhodozyma

    Bioprocess Eng.

    (1998)
  • J. Ramírez1 et al.

    Astaxanthin production by Phaffia rhodozyma in a fedbatch culture using a low cost medium feeding

    e-Gnosis

    (2006)
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    Gui-Li Jiang and Ling-Yan Zhou contributed equally to this work and should be considered co-first authors.

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