• 한국미생물·생명공학회지
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• 제49권2호
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• pp.210-216
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• 2021

본 연구에서는 CRISPR/CAS9을 이용하여 S. cerevisiae의 ACC1, ELO1, OLE1 유전자의 프로모터를 TEF1으로 교체하여 그 발현량을 증가시키고 그에 따른 에탄올에 대한 저항성과 생산량 변화를 확인하였다. 18% 에탄올이 함유된 YPD 배지에서 control을 제외하고 유전자 과발현을 일으킨 mutant 균주 모두가 24시간까지 viable하게 생존하는 것을 확인하였다. 에탄올 발효에서는 유전자 과발현 균주 모두가 에탄올 수율에서 ACC1 과발현 균주가 428.18 ± 0.29 mg/g, ELO1 과발현 균주는 416.15 ± 4.3 mg/g, OLE1 과발현 균주는 430.55 ± 6.00 mg/g에 도달하였으며, 이는 control의 수율인 400.26 ± 0.42 mg/g 보다 높은 수준에 도달하였다. 이 결과는 높은 농도의 에탄올에서 탄소 사슬이 긴 불포화지방산의 비율이 증가한다는 연구결과가 역 또한 성립한다는 것을 증명하였다. ELO1의 과발현은 elongation of fatty acid protein의 생산 증가를 불러 일으킨다. 또한 OLE1도 acylCoA desaturase 효소의 활성을 증대시킨다. TEF1이라는 strong promoter를 이용한 이번 실험에서 ELO1 과발현 균주가 OLE1 과발현 균주보다 S. cerevisiae의 에탄올 저해 감소와 발효에 긍정적인 영향을 미침을 확인하였다.

• Journal of Microbiology and Biotechnology
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• 제22권7호
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• pp.990-999
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• 2012

The microbial biosynthesis of fatty acid of lipid metabolism, which can be used as precursors for the production of fuels of chemicals from renewable carbon sources, has attracted significant attention in recent years. The regulation of fatty acid biosynthesis pathways has been mainly studied in a model prokaryote, Escherichia coli. During the recent period, global regulation of fatty acid metabolic pathways has been demonstrated in another model prokaryote, Bacillus subtilis, as well as in Streptococcus pneumonia. The goal of this study was to increase the production of long-chain fatty acids by developing recombinant E. coli strains that were improved by an elongation cycle of fatty acid synthesis (FAS). The fabB, fabG, fabZ, and fabI genes, all homologous of E. coli, were induced to improve the enzymatic activities for the purpose of overexpressing components of the elongation cycle in the FAS pathway through metabolic engineering. The ${\beta}$-oxoacyl-ACP synthase enzyme catalyzed the addition of acyl-ACP to malonyl-ACP to generate ${\beta}$-oxoacyl-ACP. The enzyme encoded by the fabG gene converted ${\beta}$-oxoacyl-ACP to ${\beta}$-hydroxyacyl-ACP, the fabZ catalyzed the dehydration of ${\beta}$-3-hydroxyacyl-ACP to trans-2-acyl-ACP, and the fabI gene converted trans-2-acyl-ACP to acyl-ACP for long-chain fatty acids. In vivo productivity of total lipids and fatty acids was analyzed to confirm the changes and effects of the inserted genes in E. coli. As a result, lipid was increased 2.16-fold higher and hexadecanoic acid was produced 2.77-fold higher in E. coli JES1030, one of the developed recombinants through this study, than those from the wild-type E. coli.

• Asian-Australasian Journal of Animal Sciences
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• 제29권11호
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• pp.1646-1652
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• 2016

Mammalian target of rapamycin complex 1 (mTORC1) is a central regulator of cell growth and metabolism and is sufficient to induce specific metabolic processes, including de novo lipid biosynthesis. Elongation of very-long-chain fatty acids 1 (ELOVL1) is a ubiquitously expressed gene and the product of which was thought to be associated with elongation of carbon (C) chain in fatty acids. In the present study, we examined the effects of rapamycin, a specific inhibitor of mTORC1, on ELOVL1 expression and docosahexaenoic acid (DHA, C22:6 n-3) synthesis in bovine mammary epithelial cells (BMECs). We found that rapamycin decreased the relative abundance of ELOVL1 mRNA, ELOVL1 expression and the level of DHA in a time-dependent manner. These data indicate that ELOVL1 expression and DHA synthesis are regulated by mTORC1 in BMECs.

• Animal Bioscience
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• 제36권10호
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• pp.1558-1567
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• 2023

Objective: Dietary green cabbage was evaluated for its impact on fatty acid synthetic ability in different adipose tissues during fattening of Wanxi White geese. Methods: A total of 256 Wanxi White geese at their 70 days were randomly allocated into 4 groups with 4 replicates and fed 0%, 15%, 30%, and 45% fresh green cabbage (relative to dry matter), respectively, in each group. Adipose tissues (subcutaneous and abdominal fat), liver and blood were collected from 4 birds in each replicate at their 70, 80, 90, and 100 days for fatty acid composition, relative gene expression and serum lipid analysis. Two-way or three-way analysis of variance was used for analysis. Results: The contents of palmitic acid (C16:0), palmitoleic acid (C16:1), linoleic acid (C18:2), and alpha-linolenic acid (C18:3) were feeding time dependently increased. The C16:0 and stearic acid (C18:0) were higher in abdominal fat, while C16:1, oleic acid (C18:1), and C18:2 were higher in subcutaneous fat. Geese fed 45% green cabbage exhibited highest level of C18:3. Geese fed green cabbage for 30 d exhibited higher level of C16:0 and C18:0 in abdominal fat, while geese fed 30% to 45% green cabbage exhibited higher C18:3 in subcutaneous fat. The expression of Acsl1 (p = 0.003) and Scd1 (p<0.0001) were decreased with green cabbage addition. Interaction between feeding time and adipose tissue affected elongation of long-chain fatty acids family member 6 (Elovl6), acyl-CoA synthetase longchain family member 1 (Acsl1), and stearoly-coA desaturase 1 (Scd1) gene expression levels (p = 0.013, p = 0.003, p = 0.005). Feeding time only affected serum lipid levels of free fatty acid and chylomicron. Higher contents of C16:0, C18:1, and C18:3 were associated with greater mRNA expression of Scd1 (p<0.0001), while higher level of C18:2 was associated with less mRNA expression of Scd1 (p<0.0001). Conclusion: Considering content of C18:2 and C18:3, 30% addition of green cabbage could be considered for fattening for 30 days in Wanxi White geese.