Function of ORFC of the polyketide synthase gene cluster on fatty acid accumulation in Schizochytrium limacinum SR21

Shi, Yanyan; Chen, Zhe; Li, Yixin; Cao, Xingyu; Yang, Lijie; Xu, Yiyuan; Li, Zhipeng; He, Ning

doi:10.1186/s13068-021-02014-9

Cited by 16 publications

(22 citation statements)

References 60 publications

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“…Knockout of ER B significantly decreased the content of PUFAs in S. limacinum SR21. Knockout of ER C reduced the content of SFAs, while overexpression of ER C led to a decrease in PUFA content and an increase in SFA content in S. limacinum SR21 (Ling et al, 2020;Shi et al, 2021). In addition, several lines of evidence have demonstrated that the addition of triclosan can inhibit the expression of ER C and lead to an increase in PUFA production (Ling et al, 2020).…”

Section: Enoyl-acp Reductase Domainmentioning

confidence: 99%

“…The heterologous expression of PUFA synthase in E. coli showed that the mutation or deletion of DH FabA -1 or substitution of DH FabA -1 with DH FabA -2 led to the loss of PUFA production capacity, and the mutation or deletion of DH FabA -2 led to a small amount of DPA production and no DHA production ( Xie et al, 2020 ). The overexpression of the DH FabA -1 domain in S. limacinum SR21 increased the content of PUFAs and total lipids ( Shi et al, 2021 ). Disruption of the DH FabA -2 domain of Schizochytrium sp.…”

Section: Function Of Domains In Eukaryotic Pufa Synthasementioning

confidence: 99%

“…There have been some studies utilizing overexpression or inactivation of domains to study their possible function and their contribution to PUFA synthesis. For example, the addition and inactivation of ACP domains have been performed in E. coli expressing eukaryotic PUFA synthase ( Hayashi et al, 2016 ); DH PKS , DH FabA -1, ER C and KR domains have been overexpressed in their native hosts ( Liu et al, 2018 ; Shi et al, 2021 ), and DH FabA -2, CLF, AT and two ER domains have been knocked out ( Li et al, 2018 ; Ren et al, 2018 ; Ling et al, 2020 ). Therefore, the functions of the remaining undetermined domains could also be analyzed in vivo using both overexpression and knockout methods in the future.…”

Section: Challenges and Future Perspectivesmentioning

confidence: 99%

“…Until now, there has been no genetic transformation method that is universally applicable for PUFA-producing eukaryotes. In thraustochytrids, electroporation is the most common strategy for transformation ( Wang et al, 2019 ; Han et al, 2020 ; Han et al, 2021 ; Rau and Ertesvag, 2021 ; Shi et al, 2021 ), and particle bombardment ( Metz et al, 2009 ) and Agrobacterium -mediated transformation ( Huang et al, 2021 ) have also been reported. To make the transformation more efficient, it is important to optimize the operating conditions according to different strains; in addition, other methods, such as bacterial conjugation ( Sreenikethanam et al, 2022 ) and cell-penetrating peptide (CPP) transport ( Rau and Ertesvag, 2021 ), should also be considered.…”

Section: Challenges and Future Perspectivesmentioning

confidence: 99%

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Deciphering and engineering the polyunsaturated fatty acid synthase pathway from eukaryotic microorganisms

Guo

Dong

Wang

et al. 2022

Front. Bioeng. Biotechnol.

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Polyunsaturated fatty acids (PUFAs) are important nutrients that play important roles in human health. In eukaryotes, PUFAs can be de novo synthesized through two independent biosynthetic pathways: the desaturase/elongase pathway and the PUFA synthase pathway. Among them, PUFAs synthesized through the PUFA synthase pathway typically have few byproducts and require fewer reduction equivalents. In the past 2 decades, numerous studies have been carried out to identify, analyze and engineer PUFA synthases from eukaryotes. These studies showed both similarities and differences between the eukaryotic PUFA synthase pathways and those well studied in prokaryotes. For example, eukaryotic PUFA synthases contain the same domain types as those in prokaryotic PUFA synthases, but the number and arrangement of several domains are different; the basic functions of same-type domains are similar, but the properties and catalytic activities of these domains are somewhat different. To further utilize the PUFA synthase pathway in microbial cell factories and improve the productivity of PUFAs, many challenges still need to be addressed, such as incompletely elucidated PUFA synthesis mechanisms and the difficult genetic manipulation of eukaryotic hosts. In this review, we provide an updated introduction to the eukaryotic PUFA synthase pathway, summarize the functions of domains and propose the possible mechanisms of the PUFA synthesis process, and then provide future research directions to further elucidate and engineer the eukaryotic PUFA synthase pathway for the maximal benefits of humans.

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Section: Enoyl-acp Reductase Domainmentioning

confidence: 99%

Section: Function Of Domains In Eukaryotic Pufa Synthasementioning

confidence: 99%

Section: Challenges and Future Perspectivesmentioning

confidence: 99%

Section: Challenges and Future Perspectivesmentioning

confidence: 99%

See 2 more Smart Citations

Deciphering and engineering the polyunsaturated fatty acid synthase pathway from eukaryotic microorganisms

Guo

Dong

Wang

et al. 2022

Front. Bioeng. Biotechnol.

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“…22 μm syringe filter (Anpel, China) to remove impurities. The carotenoids were analyzed by HPLC (1200 series, Agilent Technologies) equipped with a YMC C30 column (250 mm × 4.6 mm i.d.…”

mentioning

confidence: 99%

Modulating DHA-Producing Schizochytrium sp. toward Astaxanthin Biosynthesis via a Seamless Genome Editing System

Liu

Lin

et al. 2022

ACS Synth. Biol.

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Schizochytrium sp. is commercially used for the production of docosahexaenoic acid (DHA). Some strains of Schizochytrium sp. are also known to produce low amounts of carotenoids, including astaxanthin and β-carotene. In order to enhance the production of astaxanthin in Schizochytrium sp., we established a seamless genome editing system with a dual selection marker for rapid screening of positive transformants. By using this system, we strengthened the endogenous mevalonate pathway, enhanced the supply of geranylgeranyl diphosphate and β-carotene, upregulated endogenous β-carotene hydroxylase, and introduced the algal astaxanthin pathway. The highest astaxanthin production in the engineered Schizochytrium sp. was achieved at 8.1 mg/L (307.1 μg/g dry cell weight) under shake-flask conditions, which was 2.6-fold higher than that in the start strain. Meanwhile, the percentage of DHA to total fatty acids was not obviously affected. We then eliminated the dual selection marker by using the Cre-loxP recombination system, and the engineered strain was ready for iterative editing. The developed system could be applied to seamlessly engineer DHA-producing Schizochytrium sp. toward astaxanthin and other value-added terpenoids, which broadens the application of this strain.

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Enhancing the accumulation of lipid and docosahexaenoic acid in Schizochytrium sp. by co‐overexpression of phosphopantetheinyl transferase and ω‐3 fatty acid desaturase

Li,

Zheng,

Yang

et al. 2023

Biotechnology Journal

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Docosahexaenoic acid (DHA) as one of ω‐3 polyunsaturated fatty acids (PUFAs), plays a key role in brain development, and is widely used in food additives and the pharmaceutical industry. Schizochytrium sp. is often considered as a satisfactory strain for DHA industrialization. The aim of this study was to assess the feasibility of phosphopantetheinyl transferase (PPTase) and ω‐3 fatty acid desaturase (FAD) for regulating DHA content in Schizochytrium sp. PPTase is essential to activate the polyketide‐like synthase (PKS) pathway, which can transfer apo‐acyl‐carrier protein (apo‐ACP) into holo‐ACP, and plays a key role in DHA synthesis. Moreover, DHA and docosapentaenoic acid (DPA) are synthesized by the PKS pathway simultaneously, so high DPA synthesis limits the increase of DHA content. In addition, the detailed mechanisms of PKS pathway have not been fully elucidated, so it is difficult to improve DHA content by modifying PKS. However, ω‐3 FAD can convert DPA into DHA, and it is the most direct and effective way to increase DHA content and reduce DPA content. Based on this, we overexpressed PPTase to enhance the synthesis of DHA by the PKS pathway, overexpressed ω‐3 FAD to convert the co‐product of the PKS pathway into DHA, and co‐overexpressed PPTase and ω‐3 FAD. With these strategies, compared with wild type, the final lipid and DHA titer were 92.5 g/L and 51.5 g/L, which increased by 46.4% and 78.1% respectively. This study established an efficient DHA production strain, and provided some feasible strategies for industrial DHA production in Schizochytrium sp.This article is protected by copyright. All rights reserved

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Function of ORFC of the polyketide synthase gene cluster on fatty acid accumulation in Schizochytrium limacinum SR21

Cited by 16 publications

References 60 publications

Deciphering and engineering the polyunsaturated fatty acid synthase pathway from eukaryotic microorganisms

Deciphering and engineering the polyunsaturated fatty acid synthase pathway from eukaryotic microorganisms

Modulating DHA-Producing Schizochytrium sp. toward Astaxanthin Biosynthesis via a Seamless Genome Editing System

Enhancing the accumulation of lipid and docosahexaenoic acid in Schizochytrium sp. by co‐overexpression of phosphopantetheinyl transferase and ω‐3 fatty acid desaturase

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