对毛霉目真菌进行高通量筛选以生产低价值和高价值脂质。
High-throughput screening of Mucoromycota fungi for production of low- and high-value lipids.
作者信息
Kosa Gergely, Zimmermann Boris, Kohler Achim, Ekeberg Dag, Afseth Nils Kristian, Mounier Jerome, Shapaval Volha
机构信息
1Faculty of Science and Technology, Norwegian University of Life Sciences, P.O. Box 5003, 1432 Ås, Norway.
2Faculty of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences, P.O. Box 5003, 1432 Ås, Norway.
出版信息
Biotechnol Biofuels. 2018 Mar 14;11:66. doi: 10.1186/s13068-018-1070-7. eCollection 2018.
BACKGROUND
Mucoromycota fungi are important producers of low- and high-value lipids. is used for arachidonic acid production at industrial scale. In addition, oleaginous Mucoromycota fungi are promising candidates for biodiesel production. A critical step in the development of such biotechnological applications is the selection of suitable strains for lipid production. The aim of the present study was to use the Duetz-microtiter plate system combined with Fourier transform infrared (FTIR) spectroscopy for high-throughput screening of the potential of 100 Mucoromycota strains to produce low- and high-value lipids.
RESULTS
With this reproducible, high-throughput method, we found several promising strains for high-value omega-6 polyunsaturated fatty acid (PUFA) and biodiesel production purposes. Gamma-linolenic acid content was the highest in UBOCC-A-109196 (24.5% of total fatty acids), and VKM F-470 (24.0%). For the first time, we observed concomitant gamma-linolenic acid and alpha-linolenic acid (up to 13.0%) production in psychrophilic strains. Arachidonic acid was present the highest amount in ATCC 32222 (41.1% of total fatty acids). Low cultivation temperature (15 °C) activated the temperature sensitive ∆17 desaturase enzyme in spp., resulting in eicosapentaenoic acid production with up to 11.0% of total fatty acids in VKM F-1494. CCM-705, CCM F-539 and UBOCC-A-101347 showed very good growth (23-26 g/L) and lipid production (7.0-8.3 g/L) with high palmitic and oleic acid, and low PUFA content, which makes them attractive candidates for biodiesel production. CCM 451 had the highest total lipid content (47.2% of biomass) of all tested strains. We also demonstrated the potential of FTIR spectroscopy for high-throughput screening of total lipid content of oleaginous fungi.
CONCLUSIONS
The use of Duetz-microtiter plate system combined with FTIR spectroscopy and multivariate analysis, is a feasible approach for high-throughput screening of lipid production in Mucoromycota fungi. Several promising strains have been identified by this method for the production of high-value PUFA and biodiesel.
背景
毛霉目真菌是低价值和高价值脂质的重要生产者。被用于工业规模的花生四烯酸生产。此外，产油毛霉目真菌是生物柴油生产的有前途的候选者。开发此类生物技术应用的关键步骤是选择适合脂质生产的菌株。本研究的目的是使用杜茨微量滴定板系统结合傅里叶变换红外（FTIR）光谱对100株毛霉目菌株生产低价值和高价值脂质的潜力进行高通量筛选。
结果
通过这种可重复的高通量方法，我们发现了几种有前途的菌株，可用于生产高价值的ω-6多不饱和脂肪酸（PUFA）和生物柴油。γ-亚麻酸含量在UBOCC-A-109196中最高（占总脂肪酸的24.5%），在VKM F-470中为24.0%。我们首次在嗜冷菌株中观察到γ-亚麻酸和α-亚麻酸（高达13.0%）的同时产生。花生四烯酸在ATCC 32222中含量最高（占总脂肪酸的41.1%）。低培养温度（15°C）激活了spp.中对温度敏感的∆17去饱和酶，导致在VKM F-1494中二十碳五烯酸的产生，占总脂肪酸的11.0%。CCM-705、CCM F-539和UBOCC-A-101347表现出非常好的生长（23-26 g/L）和脂质生产（7.0-8.3 g/L），棕榈酸和油酸含量高，PUFA含量低，这使其成为生物柴油生产的有吸引力的候选者。CCM 451在所有测试菌株中总脂质含量最高（占生物量的47.2%）。我们还证明了FTIR光谱在高通量筛选产油真菌总脂质含量方面的潜力。
结论
使用杜茨微量滴定板系统结合FTIR光谱和多变量分析，是一种对毛霉目真菌脂质生产进行高通量筛选的可行方法。通过这种方法已鉴定出几种有前途的菌株，用于生产高价值PUFA和生物柴油。
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