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Main Authors: Guo, Shuilian, Liao, Shangfeng, Chen, Haixin, Xue, Chuang, Zhang, Huiming, Yue, Zhen, Zhang, Xiuqing, Fang, Xiaodong
Format: Artículo científico
Language:en
Published: ACS synthetic biology 2025
Subjects:
Online Access:https://pubmed.ncbi.nlm.nih.gov/40875908/
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author Guo, Shuilian
Liao, Shangfeng
Chen, Haixin
Xue, Chuang
Zhang, Huiming
Yue, Zhen
Zhang, Xiuqing
Fang, Xiaodong
author_facet Guo, Shuilian
Liao, Shangfeng
Chen, Haixin
Xue, Chuang
Zhang, Huiming
Yue, Zhen
Zhang, Xiuqing
Fang, Xiaodong
Guo, Shuilian
Liao, Shangfeng
Chen, Haixin
Xue, Chuang
Zhang, Huiming
Yue, Zhen
Zhang, Xiuqing
Fang, Xiaodong
collection PubMed - marine biology
contents Enhancing Astaxanthin Biosynthesis in Synthetic Yeast Through Combinatorial Metabolic Engineering and Genome-Scale Evolution. Guo, Shuilian Liao, Shangfeng Chen, Haixin Xue, Chuang Zhang, Huiming Yue, Zhen Zhang, Xiuqing Fang, Xiaodong Metabolic Engineering Xanthophylls Saccharomyces cerevisiae Mixed Function Oxygenases Oxygenases Directed Molecular Evolution Synthetic Biology Astaxanthin, a high-value keto-carotenoid with exceptional antioxidant capacity, has significant commercial potential for industrial applications. Microbial biosynthesis via engineered synthetic yeast presents an environmentally sustainable production platform. In this study, we developed a multistrategy optimization framework to enhance astaxanthin biosynthesis in synthetic yeast. Our systematic approach initiated with the construction of a de novo astaxanthin pathway in synthetic yeast strain 2369R, achieving a baseline production of 0.11 mg/L. Through rigorous screening of heterologous enzymes, we identified optimal variants of β-carotene hydroxylase () and ketolase () that increased the titer to 0.65 mg/L. Subsequently, the combined enhancement of MVA pathway flux (via overexpression) and lipid metabolism regulation (through overexpression) synergistically boosted astaxanthin production to 2.59 mg/L. Through combinatorial implementation of genome-scale diversification using the Synthetic Chromosome Rearrangement and Modification by LoxP-mediated Evolution (SCRaMbLE) system coupled with an absorption-based semi-high-throughput screening platform (/), we successfully isolated an elite mutant strain, YgM97, that achieved 6.85 mg/L astaxanthin production in shake-flask culture. This represents a remarkable 61.27-fold enhancement compared with the parental strain. Transcriptomic and genomic analyses subsequently revealed the potential molecular mechanisms underlying this significant yield improvement. Collectively, this study demonstrates the powerful synergy between rational metabolic engineering and randomized genome evolution, providing a novel paradigm for high-value compound biosynthesis in a microbial chassis.
format Artículo científico
id pubmed_40875908
institution PubMed
language en
publishDate 2025
publisher ACS synthetic biology
record_format pubmed
spellingShingle Enhancing Astaxanthin Biosynthesis in Synthetic Yeast Through Combinatorial Metabolic Engineering and Genome-Scale Evolution.
Guo, Shuilian
Liao, Shangfeng
Chen, Haixin
Xue, Chuang
Zhang, Huiming
Yue, Zhen
Zhang, Xiuqing
Fang, Xiaodong
Metabolic Engineering
Xanthophylls
Saccharomyces cerevisiae
Mixed Function Oxygenases
Oxygenases
Directed Molecular Evolution
Synthetic Biology
Enhancing Astaxanthin Biosynthesis in Synthetic Yeast Through Combinatorial Metabolic Engineering and Genome-Scale Evolution. Guo, Shuilian Liao, Shangfeng Chen, Haixin Xue, Chuang Zhang, Huiming Yue, Zhen Zhang, Xiuqing Fang, Xiaodong Metabolic Engineering Xanthophylls Saccharomyces cerevisiae Mixed Function Oxygenases Oxygenases Directed Molecular Evolution Synthetic Biology Astaxanthin, a high-value keto-carotenoid with exceptional antioxidant capacity, has significant commercial potential for industrial applications. Microbial biosynthesis via engineered synthetic yeast presents an environmentally sustainable production platform. In this study, we developed a multistrategy optimization framework to enhance astaxanthin biosynthesis in synthetic yeast. Our systematic approach initiated with the construction of a de novo astaxanthin pathway in synthetic yeast strain 2369R, achieving a baseline production of 0.11 mg/L. Through rigorous screening of heterologous enzymes, we identified optimal variants of β-carotene hydroxylase () and ketolase () that increased the titer to 0.65 mg/L. Subsequently, the combined enhancement of MVA pathway flux (via overexpression) and lipid metabolism regulation (through overexpression) synergistically boosted astaxanthin production to 2.59 mg/L. Through combinatorial implementation of genome-scale diversification using the Synthetic Chromosome Rearrangement and Modification by LoxP-mediated Evolution (SCRaMbLE) system coupled with an absorption-based semi-high-throughput screening platform (/), we successfully isolated an elite mutant strain, YgM97, that achieved 6.85 mg/L astaxanthin production in shake-flask culture. This represents a remarkable 61.27-fold enhancement compared with the parental strain. Transcriptomic and genomic analyses subsequently revealed the potential molecular mechanisms underlying this significant yield improvement. Collectively, this study demonstrates the powerful synergy between rational metabolic engineering and randomized genome evolution, providing a novel paradigm for high-value compound biosynthesis in a microbial chassis.
title Enhancing Astaxanthin Biosynthesis in Synthetic Yeast Through Combinatorial Metabolic Engineering and Genome-Scale Evolution.
topic Metabolic Engineering
Xanthophylls
Saccharomyces cerevisiae
Mixed Function Oxygenases
Oxygenases
Directed Molecular Evolution
Synthetic Biology
url https://pubmed.ncbi.nlm.nih.gov/40875908/