Photoautotrophic production of macular pigment in a Chlamydomonas reinhardtii strain generated by using DNA-free CRISPR-Cas9 RNP-mediated mutagenesis

Kwangryul Baek, Jihyeon Yu, Jooyeon Jeong, Sang Jun Sim, Sangsu Bae, Eon Seon Jin

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Lutein and zeaxanthin are dietary carotenoids reported to be protective against age-related macular degeneration. Recently, the green alga Chlamydomonas reinhardtii has received attention as a photosynthetic cell factory, but the potential of this alga for carotenoid production has not yet been evaluated. In this study, we selected the C. reinhardtii CC-4349 strain as the best candidate among seven laboratory strains tested for carotenoid production. A knock-out mutant of the zeaxanthin epoxidase gene induced by preassembled DNA-free CRISPR-Cas9 ribonucleoproteins in the CC-4349 strain had a significantly higher zeaxanthin content (56-fold) and productivity (47-fold) than the wild type without the reduction in lutein level. Furthermore, we produced eggs fortified with lutein (2-fold) and zeaxanthin (2.2-fold) by feeding hens a diet containing the mutant. Our results clearly demonstrate the possibility of cost-effective commercial use of microalgal mutants induced by DNA-free CRISPR-Cas9 ribonucleoproteins in algal biotechnology for the production of high-value products.

Original languageEnglish
Pages (from-to)719-728
Number of pages10
JournalBiotechnology and Bioengineering
Volume115
Issue number3
DOIs
Publication statusPublished - 2018 Mar 1

Fingerprint

Clustered Regularly Interspaced Short Palindromic Repeats
Chlamydomonas reinhardtii
Lutein
Mutagenesis
Carotenoids
Pigments
DNA
Ribonucleoproteins
Algae
Chlorophyta
Macular Degeneration
Biotechnology
Nutrition
Eggs
Industrial plants
Genes
Productivity
Diet
Costs and Cost Analysis
Macular Pigment

Keywords

  • algal biotechnology
  • Chlamydomonas reinhardtii
  • CRISPR-Cas9
  • lutein
  • zeaxanthin

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

Cite this

Photoautotrophic production of macular pigment in a Chlamydomonas reinhardtii strain generated by using DNA-free CRISPR-Cas9 RNP-mediated mutagenesis. / Baek, Kwangryul; Yu, Jihyeon; Jeong, Jooyeon; Sim, Sang Jun; Bae, Sangsu; Jin, Eon Seon.

In: Biotechnology and Bioengineering, Vol. 115, No. 3, 01.03.2018, p. 719-728.

Research output: Contribution to journalArticle

@article{595e7900745b4f028cd43ef369425916,
title = "Photoautotrophic production of macular pigment in a Chlamydomonas reinhardtii strain generated by using DNA-free CRISPR-Cas9 RNP-mediated mutagenesis",
abstract = "Lutein and zeaxanthin are dietary carotenoids reported to be protective against age-related macular degeneration. Recently, the green alga Chlamydomonas reinhardtii has received attention as a photosynthetic cell factory, but the potential of this alga for carotenoid production has not yet been evaluated. In this study, we selected the C. reinhardtii CC-4349 strain as the best candidate among seven laboratory strains tested for carotenoid production. A knock-out mutant of the zeaxanthin epoxidase gene induced by preassembled DNA-free CRISPR-Cas9 ribonucleoproteins in the CC-4349 strain had a significantly higher zeaxanthin content (56-fold) and productivity (47-fold) than the wild type without the reduction in lutein level. Furthermore, we produced eggs fortified with lutein (2-fold) and zeaxanthin (2.2-fold) by feeding hens a diet containing the mutant. Our results clearly demonstrate the possibility of cost-effective commercial use of microalgal mutants induced by DNA-free CRISPR-Cas9 ribonucleoproteins in algal biotechnology for the production of high-value products.",
keywords = "algal biotechnology, Chlamydomonas reinhardtii, CRISPR-Cas9, lutein, zeaxanthin",
author = "Kwangryul Baek and Jihyeon Yu and Jooyeon Jeong and Sim, {Sang Jun} and Sangsu Bae and Jin, {Eon Seon}",
year = "2018",
month = "3",
day = "1",
doi = "10.1002/bit.26499",
language = "English",
volume = "115",
pages = "719--728",
journal = "Biotechnology and Bioengineering",
issn = "0006-3592",
publisher = "Wiley-VCH Verlag",
number = "3",

}

TY - JOUR

T1 - Photoautotrophic production of macular pigment in a Chlamydomonas reinhardtii strain generated by using DNA-free CRISPR-Cas9 RNP-mediated mutagenesis

AU - Baek, Kwangryul

AU - Yu, Jihyeon

AU - Jeong, Jooyeon

AU - Sim, Sang Jun

AU - Bae, Sangsu

AU - Jin, Eon Seon

PY - 2018/3/1

Y1 - 2018/3/1

N2 - Lutein and zeaxanthin are dietary carotenoids reported to be protective against age-related macular degeneration. Recently, the green alga Chlamydomonas reinhardtii has received attention as a photosynthetic cell factory, but the potential of this alga for carotenoid production has not yet been evaluated. In this study, we selected the C. reinhardtii CC-4349 strain as the best candidate among seven laboratory strains tested for carotenoid production. A knock-out mutant of the zeaxanthin epoxidase gene induced by preassembled DNA-free CRISPR-Cas9 ribonucleoproteins in the CC-4349 strain had a significantly higher zeaxanthin content (56-fold) and productivity (47-fold) than the wild type without the reduction in lutein level. Furthermore, we produced eggs fortified with lutein (2-fold) and zeaxanthin (2.2-fold) by feeding hens a diet containing the mutant. Our results clearly demonstrate the possibility of cost-effective commercial use of microalgal mutants induced by DNA-free CRISPR-Cas9 ribonucleoproteins in algal biotechnology for the production of high-value products.

AB - Lutein and zeaxanthin are dietary carotenoids reported to be protective against age-related macular degeneration. Recently, the green alga Chlamydomonas reinhardtii has received attention as a photosynthetic cell factory, but the potential of this alga for carotenoid production has not yet been evaluated. In this study, we selected the C. reinhardtii CC-4349 strain as the best candidate among seven laboratory strains tested for carotenoid production. A knock-out mutant of the zeaxanthin epoxidase gene induced by preassembled DNA-free CRISPR-Cas9 ribonucleoproteins in the CC-4349 strain had a significantly higher zeaxanthin content (56-fold) and productivity (47-fold) than the wild type without the reduction in lutein level. Furthermore, we produced eggs fortified with lutein (2-fold) and zeaxanthin (2.2-fold) by feeding hens a diet containing the mutant. Our results clearly demonstrate the possibility of cost-effective commercial use of microalgal mutants induced by DNA-free CRISPR-Cas9 ribonucleoproteins in algal biotechnology for the production of high-value products.

KW - algal biotechnology

KW - Chlamydomonas reinhardtii

KW - CRISPR-Cas9

KW - lutein

KW - zeaxanthin

UR - http://www.scopus.com/inward/record.url?scp=85041078024&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85041078024&partnerID=8YFLogxK

U2 - 10.1002/bit.26499

DO - 10.1002/bit.26499

M3 - Article

C2 - 29150930

AN - SCOPUS:85041078024

VL - 115

SP - 719

EP - 728

JO - Biotechnology and Bioengineering

JF - Biotechnology and Bioengineering

SN - 0006-3592

IS - 3

ER -