Development of a Transformation System for Nitratireductor sp.
Development of a Transformation System for Nitratireductor sp.
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DOI：
10.1007/s10126-023-10198-4
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发表时间：
2023-10
影响因子：
3
通讯作者：
Okamura, Yoshiko
中科院分区：
文献类型：
--
作者：
Maeda, Hiroto;Hirata, Yuto;Takahashi, Hirokazu;Watanabe, Kenshi;Aki, Tsunehiro;Okamura, Yoshiko
硝酸还原细菌。OM-1可以积累丁烯酸，丁烯酸是一种用于化工产品的短链不饱和羧酸。到目前为止，基于比较转录组分析，我们预测了硫代酯酶基因TE，作为丁烯酸生物合成的候选基因。为了确定TE的功能，对硝酸还原细菌的基因转移系统进行了研究。OM-1是必需的。因此，在本研究中，我们选择了电穿孔转化系统和宿主范围较广的pRK415，并对转化条件进行了优化。结果表明，在22.5kV/cm时，最高转化效率为7.9DNA× × 10~4个菌落/微克DNA。此外，通过插入TE构建了表达载体pRK415-TE，并利用电穿孔法成功地将其转化到菌株OM-1中。重组OM-1菌株的丁烯酸产量为26.7 mg/g干重，比含有空载体的转化子提高了254%。这是首次报道了硝化还原细菌的基因转移系统，表明TE基因负责丁烯酸的产生。网上版载有补充材料，可在10.1007/s10126-023-10198-4查阅。
Nitratireductor sp. OM-1 can accumulate butenoic acid, which is a short-chain unsaturated carboxylic acid utilized for chemical products. So far, we have predicted the thioesterase gene, te, as a candidate gene for butenoic acid biosynthesis, based on comparative transcriptome analysis. To confirm the function of te, the gene transfer system in Nitratireductor sp. OM-1 was required. Thus, in this study, we used electroporation as a transformation system and pRK415, a broad host range plasmid, and optimized the conditions. As a result, a maximum transformation efficiency of 7.9 × 104 colonies/µg DNA was obtained at 22.5 kV/cm. Moreover, an expression vector, pRK415-te, was constructed by insertion of te, which was successfully transferred into strain OM-1, using electroporation. The recombinant OM-1 strain produced butenoic acid at 26.7 mg/g of dried cell weight, which was a 254% increase compared to transformants harboring an empty vector. This is the first report of a gene transfer system for Nitratireductor sp., which showed that the te gene was responsible for butenoic acid production. The online version contains supplementary material available at 10.1007/s10126-023-10198-4.
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影响因子：
11.1
作者：
Mamat, Mohd Rahimi Zakaria;Ariffin, Hidayah;Zahari, Mior Ahmad Khushairi Mohd
通讯作者：
Zahari, Mior Ahmad Khushairi Mohd
影响因子：
11.4
作者：
Okamura, Yoshiko;Nakai, Shota;Matsumoto, Mitsufumi
通讯作者：
Matsumoto, Mitsufumi
影响因子：
3.2
作者：
CALVIN, NM;HANAWALT, PC
通讯作者：
HANAWALT, PC
影响因子：
4
作者：
Mañas, P;Pagán, R
通讯作者：
Pagán, R
影响因子：
5
作者：
Liu, Xiping;Yu, Haiying;Zhu, Kun
通讯作者：
Zhu, Kun