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Engineering the oleaginous yeast Yarrowia lipolytica to produce the aroma compound β-ionone

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    • 화장품용 기능성소재
      1. 기능성
      2. 계면활성제⁄증점제
      3. 기타
    • 의료용 화학소재
      1. 건강보조식품
논문

Engineering the oleaginous yeast Yarrowia lipolytica to produce the aroma compound β-ionone

학술지

Microbial cell factories

저자명

Czajka, Jeffrey J.; Nathenson, Justin A.; Benites, Veronica T.; Baidoo, Edward E. K.; Cheng, Qianshun; Wang, Yechun; Tang, Yinjie J.

초록

<P><B>Background</B></P><P>&beta;-Ionone is a fragrant terpenoid that generates a pleasant floral scent and is used in diverse applications as a cosmetic and flavoring ingredient. A growing consumer desire for natural products has increased the market demand for natural &beta;-ionone. To date, chemical extraction from plants remains the main approach for commercial natural &beta;-ionone production. Unfortunately, changing climate and geopolitical issues can cause instability in the &beta;-ionone supply chain. Microbial fermentation using generally recognized as safe (GRAS) yeast offers an alternative method for producing natural &beta;-ionone. <I>Yarrowia lipolytica</I> is an attractive host due to its oleaginous nature, established genetic tools, and large intercellular pool size of acetyl-CoA (the terpenoid backbone precursor).</P><P><B>Results</B></P><P>A push&#x2013;pull strategy via genome engineering was applied to a <I>Y. lipolytica</I> PO1f derived strain. Heterologous and native genes in the mevalonate pathway were overexpressed to push production to the terpenoid backbone geranylgeranyl pyrophosphate, while the <I>carB</I> and biofunction <I>carRP</I> genes from <I>Mucor circinelloides</I> were introduced to pull flux towards &beta;-carotene (i.e., ionone precursor). Medium tests combined with machine learning based data analysis and <SUP>13</SUP>C metabolite labeling investigated influential nutrients for the &beta;-carotene strain that achieved > 2.5&nbsp;g/L &beta;-carotene in a rich medium. Further introduction of the carotenoid cleavage dioxygenase 1 (CCD1) from <I>Osmanthus fragrans</I> resulted in the &beta;-ionone production. Utilization of in situ dodecane trapping avoided ionone loss from vaporization (with recovery efficiencies of ~ 76%) during fermentation operations, which resulted in titers of 68&nbsp;mg/L &beta;-ionone in shaking flasks and 380&nbsp;mg/L in a 2&nbsp;L fermenter. Both &beta;-carotene medium tests and &beta;-ionone fermentation outcomes indicated the last enzymatic step CCD1 (rather than acetyl-CoA supply) as the key bottleneck.</P><P><B>Conclusions</B></P><P>We engineered a GRAS <I>Y. lipolytica</I> platform for sustainable and economical production of the natural aroma &beta;-ionone. Although &beta;-carotene could be produced at high titers by <I>Y. lipolytica</I>, the synthesis of &beta;-ionone was relatively poor, possibly due to low CCD1 activity and non-specific CCD1 cleavage of &beta;-carotene. In addition, both &beta;-carotene and &beta;-ionone strains showed decreased performances after successive sub-cultures. For industrial application, &beta;-ionone fermentation efforts should focus on both CCD enzyme engineering and strain stability improvement.</P><P><B>Electronic supplementary material</B></P><P>The online version of this article (10.1186/s12934-018-0984-x) contains supplementary material, which is available to authorized users.</P>

발행연도

2018

발행기관

BioMed Central

라이선스

cc-by

ISSN

1475-2859

17

페이지

pp.136

주제어

13C labeling; Terpenoid; Acetyl-CoA; β-carotene; Machine learning; Fed-batch fermentation; Strain stability

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1 2023-12-11
2 2023-12-11

논문; 2018-09-01

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