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Induced high-yield production of zeaxanthin, lutein, and #-carotene by a mutant of Chlorella zofingiensis Weiping Huang, Yan Lin, Mingxia He, Yuhao Gong, and Junchao Huang J. Agric. Food Chem., Just Accepted Manuscript • DOI: 10.1021/acs.jafc.7b05400 • Publication Date (Web): 10 Jan 2018 Downloaded from http://pubs.acs.org on January 10, 2018
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Journal of Agricultural and Food Chemistry
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Induced high-yield production of zeaxanthin, lutein, and β-carotene by a
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mutant of Chlorella zofingiensis
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Weiping Huang, Yan Lin, Mingxia He, Yuhao Gong, Junchao Huang*
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Department of Economic Plants and Biotechnology, Yunnan Key Laboratory for Wild Plant
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Resources, Kunming Institute of Botany, Chinese Academy of Sciences, Kunming 650201
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*corresponding author, E-mail address:
[email protected] 11
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Journal of Agricultural and Food Chemistry
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ABSTRACT
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Natural resources of zeaxanthin are extremely limited. A Chlorella zofingiensis mutant (CZ-
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bkt1), which could accumulate high amounts of zeaxanthin, was generated and characterized.
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CZ-bkt1 was achieved by treating the algal cells with a chemical mutagen followed by a
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colour-based colony screening approach. CZ-bkt1 was found to consist of a dysfunctional
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carotenoid ketolase, leading to the accumulation of zeaxanthin rather than its downstream
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ketocarotenoid astaxanthin. Light irradiation, glucose, NaCl, and nitrogen deficiency all
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induced CZ-bkt1 to accumulate zeaxanthin. CZ-bkt1 accumulated zeaxanthin up to
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7.00±0.82 mg/g when induced by high light irradiation and nitrogen deficiency, and
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36.79±2.23 mg/L by additional feeding with glucose. Furthermore, in addition to zeaxanthin,
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CZ-bkt1 also accumulated high amounts of β-carotene (7.18±0.72 mg/g or 34.64±1.39 mg/L)
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and lutein (13.81±1.23 mg/g or 33.97±2.61 mg/L). CZ-bkt1 is the sole species up to date with
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the ability to accumulate high amounts of the three carotenoids that are essential for human
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health.
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KEYWORDS
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Carotenoid; Chlorella zofingiensis; Zeaxanthin; Lutein; Microalgae
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Journal of Agricultural and Food Chemistry
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1. INTRODUCTION
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Carotenoids are yellow to red pigments synthesized by all photosynthetic organisms. Lutein,
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β-carotene, and zeaxanthin are three typical carotenoids with important biological functions.
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They are essential nutrients for human health in that β-carotene serves as provitamin A, while
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lutein and zeaxanthin are important pigments in the retina 1, 2. Vitamin A deficiency is a
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public health problem in developing countries, which has prompted great efforts toward the
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biofortification of plant-derived foods with provitamin A carotenoids 3, 4. High dietary intake
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of zeaxanthin and lutein proved to decrease the risk of age-related macular degeneration, the
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main cause of blindness of elder people 5. Zeaxanthin and lutein have been used as feed
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additives and colorants in the food industry for animals 6, 7. Studies have also found that
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zeaxanthin could reduce the risks of cataract, cancer and low-density lipoprotein oxidation 8, 9.
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At least 2 mg/day of zeaxanthin was recommended by European Commission for adult health
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10, 11
.
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Microalgae have been exploited for producing high-value carotenoids, e.g. astaxanthin
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from Haematococcus pluvialis 12, β-carotene from Dunaliella bardawill 13 and lutein from
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Chlorella species 14. Compared with other carotenoids, zeaxanthin levels are extremely low,
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e.g.,