调节产生 DHA 的裂殖壶菌属 sp。通过无缝基因组编辑系统进行虾青素生物合成,ACS Synthetic Biology

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调节产生 DHA 的裂殖壶菌属 sp。通过无缝基因组编辑系统进行虾青素生物合成
ACS Synthetic Biology ( IF 3.7 ) Pub Date : 2022-12-01 , DOI: 10.1021/acssynbio.2c00490
Peng-Yang Liu ₁ , Gang Li ₁ , Chu-Bin Lin ₁ , Jun-Jie Wu ₁ , Shan Jiang ₁ , Feng-Hong Huang _{1,

2,

3} , Xia Wan _{1,

2,

3}

Affiliation

裂殖壶菌属商业上用于生产二十二碳六烯酸 (DHA)。一些菌株Schizochytrium sp。还已知会产生少量的类胡萝卜素，包括虾青素和 β-胡萝卜素。为了提高Schizochytrium sp. 虾青素的产量，我们建立了一个带有双重选择标记的无缝基因组编辑系统，用于快速筛选阳性转化子。通过使用该系统，我们加强了内源性甲羟戊酸途径，增加了香叶基香叶基二磷酸和β-胡萝卜素的供应，上调了内源性β-胡萝卜素羟化酶，并引入了藻类虾青素途径。经改造的裂殖壶菌中虾青素产量最高sp。在摇瓶条件下达到 8.1 mg/L（307.1 μg/g 细胞干重），比起始菌株高 2.6 倍。同时，DHA占总脂肪酸的百分比没有受到明显影响。然后，我们使用 Cre-loxP 重组系统消除了双重选择标记，工程菌株已准备好进行迭代编辑。所开发的系统可用于无缝设计产生 DHA 的裂殖壶菌属。对虾青素和其他增值萜类化合物，这拓宽了该菌株的应用。

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

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.

更新日期：2022-12-01

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