期刊
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
卷 67, 期 4, 页码 1072-1080出版社
AMER CHEMICAL SOC
DOI: 10.1021/acs.jafc.8b05003
关键词
astaxanthin; beta-carotene hydroxylase; directed coevolution; temperature-responsive regulation; high-density fermentation
资金
- Zhejiang Provincial Natural Science Foundation of China [LY18B060001]
- Natural Science Foundation of China [21576234, 21776244]
Because it is an outstanding antioxidant with wide applications, biotechnological production of astaxanthin has attracted increasing research interest. However, the astaxanthin titer achieved to date is still rather low, attributed to the poor efficiency of beta-carotene ketolation and hydroxylation, as well as the adverse effect of astaxanthin accumulation on cell growth. To address these problems, we constructed an efficient astaxanthin-producing Saccharomyces cerevisiae strain by combining protein engineering and dynamic metabolic regulation. First, superior mutants of fi-carotene ketolase and beta-carotene hydroxylase were obtained by directed coevolution to accelerate the conversion of beta-carotene to astaxanthin. Subsequently, the Gal4M9-based temperature-responsive regulation system was introduced to separate astaxanthin production from cell growth. Finally, 235 mg/L of (3S,3'S)-astaxanthin was produced by two-stage, high-density fermentation. This study demonstrates the power of combining directed coevolution and temperature-responsive regulation in astaxanthin biosynthesis and may provide methodological reference for biotechnological production of other value-added chemicals.
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