Metabolic Engineering of Chlamydomonas reinhardtii for Enhanced β-Carotene and Lutein Production

2019 ◽  
Vol 190 (4) ◽  
pp. 1457-1469 ◽  
Author(s):  
Jayant Pralhad Rathod ◽  
Chaitali Vira ◽  
Arvind M. Lali ◽  
Gunjan Prakash
Keyword(s):  
Metabolic Engineering ◽  
Chlamydomonas Reinhardtii ◽  
Β Carotene ◽  
Lutein Production

Expression of β-carotene hydroxylase gene (crtR-B) from the green alga Haematococcus pluvialis in chloroplasts of Chlamydomonas reinhardtii

2007 ◽  
Vol 19 (4) ◽  
pp. 347-355 ◽  
Author(s):  
Cong-Ping Tan ◽  
Fang-Qing Zhao ◽  
Zhong-Liang Su ◽  
Cheng-Wei Liang ◽  
Song Qin
Keyword(s):  
Chlamydomonas Reinhardtii ◽  
Green Alga ◽  
Haematococcus Pluvialis ◽  
Hydroxylase Gene ◽  
Β Carotene

Metabolic engineering of β-carotene biosynthesis in Yarrowia lipolytica

2020 ◽  
Vol 42 (6) ◽  
pp. 945-956 ◽  
Author(s):  
Xin-Kai Zhang ◽  
Dan-Ni Wang ◽  
Jun Chen ◽  
Zhi-Jie Liu ◽  
Liu-Jing Wei ◽  
...  
Keyword(s):  
Metabolic Engineering ◽  
Yarrowia Lipolytica ◽  
Β Carotene

scholarly journals Metabolic Engineering of Saccharomyces cerevisiae for Astaxanthin Production and Oxidative Stress Tolerance

2009 ◽  
Vol 75 (22) ◽  
pp. 7205-7211 ◽  
Author(s):  
Ken Ukibe ◽  
Keisuke Hashida ◽  
Nobuyuki Yoshida ◽  
Hiroshi Takagi
Keyword(s):  
Oxidative Stress ◽  
Saccharomyces Cerevisiae ◽  
Cytochrome P450 ◽  
Metabolic Engineering ◽  
Stress Tolerance ◽  
Yeast Cells ◽  
Cytochrome P450 Enzyme ◽  
Oxidative Stress Tolerance ◽  
Astaxanthin Production ◽  
Β Carotene

ABSTRACT The red carotenoid astaxanthin possesses higher antioxidant activity than other carotenoids and has great commercial potential for use in the aquaculture, pharmaceutical, and food industries. In this study, we produced astaxanthin in the budding yeast Saccharomyces cerevisiae by introducing the genes involved in astaxanthin biosynthesis of carotenogenic microorganisms. In particular, expression of genes of the red yeast Xanthophyllomyces dendrorhous encoding phytoene desaturase (crtI product) and bifunctional phytoene synthase/lycopene cyclase (crtYB product) resulted in the accumulation of a small amount of β-carotene in S. cerevisiae. Overexpression of geranylgeranyl pyrophosphate (GGPP) synthase from S. cerevisiae (the BTS1 gene product) increased the intracellular β-carotene levels due to the accelerated conversion of farnesyl pyrophosphate to GGPP. Introduction of the X. dendrorhous crtS gene, encoding astaxanthin synthase, assumed to be the cytochrome P450 enzyme, did not lead to astaxanthin production. However, coexpression of CrtS with X. dendrorhous CrtR, a cytochrome P450 reductase, resulted in the accumulation of a small amount of astaxanthin. In addition, the β-carotene-producing yeast cells transformed by the bacterial genes crtW and crtZ, encoding β-carotene ketolase and hydroxylase, respectively, also accumulated astaxanthin and its intermediates, echinenone, canthaxanthin, and zeaxanthin. Interestingly, we found that these ketocarotenoids conferred oxidative stress tolerance on S. cerevisiae cells. This metabolic engineering has potential for overproduction of astaxanthin and breeding of novel oxidative stress-tolerant yeast strains.

scholarly journals A teaching protocol demonstrating the use of EasyClone and CRISPR/Cas9 for metabolic engineering of Saccharomyces cerevisiae and Yarrowia lipolytica

2019 ◽  
Author(s):  
N Milne ◽  
L R R Tramontin ◽  
I Borodina
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Metabolic Engineering ◽  
Yarrowia Lipolytica ◽  
Yeast Species ◽  
Production Capacity ◽  
Value Added ◽  
Phenotypic Screen ◽  
Rate Limiting ◽  
Β Carotene

ABSTRACT We present a teaching protocol suitable for demonstrating the use of EasyClone and CRISPR/Cas9 for metabolic engineering of industrially relevant yeasts Saccharomyces cerevisiae and Yarrowia lipolytica, using β-carotene production as a case study. The protocol details all steps required to generate DNA parts, transform and genotype yeast, and perform a phenotypic screen to determine β-carotene production. The protocol is intended to be used as an instruction manual for a two-week practical course aimed at MSc and PhD students. The protocol details all necessary steps for students to engineer yeast to produce β-carotene and serves as a practical introduction to the principles of metabolic engineering including the concepts of boosting native precursor supply and alleviating rate-limiting steps. It also highlights key differences in the metabolism and heterologous production capacity of two industrially relevant yeast species. The protocol is divided into daily experiments covering a two week period and provides detailed instructions for every step meaning this protocol can be used ‘as is’ for a teaching course or as a case study for how yeast can be engineered to produce value-added molecules.

scholarly journals Metabolic engineering of β-carotene in orange fruit increases itsin vivoantioxidant properties

2013 ◽  
Vol 12 (1) ◽  
pp. 17-27 ◽  
Author(s):  
Elsa Pons ◽  
Berta Alquézar ◽  
Ana Rodríguez ◽  
Patricia Martorell ◽  
Salvador Genovés ◽  
...  
Keyword(s):  
Metabolic Engineering ◽  
Orange Fruit ◽  
Β Carotene

Enhanced Lutein Production in Chlamydomonas reinhardtii by Overexpression of the Lycopene Epsilon Cyclase Gene

2021 ◽  
Author(s):  
Saki Tokunaga ◽  
Daichi Morimoto ◽  
Takahisa Koyama ◽  
Yuki Kubo ◽  
Mai Shiroi ◽  
...  
Keyword(s):  
Chlamydomonas Reinhardtii ◽  
Lutein Production
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