金核霉素生物合成途径的体内研究和关键代谢物鉴定

doi:10.6023/cjoc202301012

有机化学 ›› 2023, Vol. 43 ›› Issue (7): 2561-2566.DOI: 10.6023/cjoc202301012 上一篇下一篇

研究简报

金核霉素生物合成途径的体内研究和关键代谢物鉴定

王飞^a, 金文兵^b, 侯现锋^b, 唐功利^a^,^b^,^*(), 潘海学^a^,^b^,^*()

^a国科大杭州高等研究院化学与材料科学学院杭州 310024
^b中国科学院上海有机化学研究所上海 200032

收稿日期:2022-11-11 修回日期:2023-02-24 发布日期:2023-03-17
通讯作者: 唐功利, 潘海学
基金资助:
国家自然科学基金(32070065)

In vivo Studies on the Biosynthetic Pathway of Aureonuclemycin and Identification of Key Metabolites

Fei Wang^a, Wenbing Jin^b, Xianfeng Hou^b, Gongli Tang^a^,^b(), Haixue Pan^a^,^b()

^aSchool of Chemistry and Materials Science, Hangzhou Institute for Advanced Study, University of Chinese Academy of Sciences, Hangzhou 310024
^bShanghai Institute of Organic Chemistry, Chinese Academy of Sciences, Shanghai 200032

Received:2022-11-11 Revised:2023-02-24 Published:2023-03-17
Contact: Gongli Tang, Haixue Pan
Supported by:
National Natural Science Foundation of China(32070065)

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摘要/Abstract

在前期获得金核霉素生物合成基因簇的基础上, 通过同框删除方法构建了全部基因敲除突变株, 确定了在金核霉素生物合成中发挥作用的关键基因. 通过突变株和野生型菌株发酵, 得到了两个关键的金核霉素类化合物. 根据体内基因敲除实验、突变株代谢物分析、关键化合物鉴定结果, 提出了金核霉素可能的生物合成途径.

关键词: 核苷类抗生素, 生物合成, 金核霉素

On the basis of aureonuclemycin biosynthetic gene cluster obtained in the previous work, in-frame markerless deletion mutants of all genes were constructed, and the key genes that play a role in the biosynthesis of aureonuclemycin were identified. Two key aureonuclemycin-like compounds were successfully obtained from mutant and wild-type strains. According to the results of in vivo experiments, metabolite analysis and identification of key compounds, the possible biosynthetic pathway of aureonuclemycin was proposed.

Key words: nucleoside antibiotic, biosynthesis, aureonuclemycin

引用此文

王飞, 金文兵, 侯现锋, 唐功利, 潘海学. 金核霉素生物合成途径的体内研究和关键代谢物鉴定[J]. 有机化学, 2023, 43(7): 2561-2566.

Fei Wang, Wenbing Jin, Xianfeng Hou, Gongli Tang, Haixue Pan. In vivo Studies on the Biosynthetic Pathway of Aureonuclemycin and Identification of Key Metabolites[J]. Chinese Journal of Organic Chemistry, 2023, 43(7): 2561-2566.

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参考文献 6

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No.	δ_C	δ_H, Mult. (J in Hz)	HSQC	¹H-¹H COSY	HMBC	NOESY
2	154.8	7.84, s	C-2	—	C-4, C-6	H-2'
4	150.2	—	—	—	—	—
5	120.0	—	—	—	—	—
6	157.1	—	—	—	—	—
8	141.2	7.88, s	C-8	—	C-4, C-5	H-2', H-6', H-8'
1'	92.4	5.79, s	C-1'	H-2'	C-4, C-8, C-2', C-3'	H-2', H-3', H-4'
2'	82.1	4.53, s	C-2'	H-1', H-3'	C-1', C-3', C-4'	H-1', H-3', H-6'
3'	77.4	4.32, s	C-3'	H-2', H-4'	C-1', C-2', C-4'	H-1', H-2', H-4', H-5'_a,b
4'	81.2	4.62, s	C-4'	H-3', H-5'_a,b	C-3', C-6'	H-1', H-3', H-5'_a,b
5'_a,b	27.1	2.22~2.34, m	C-5'	H-4', H-6'	C-3', C-4', C-6', C-7'	H-3', H-4', H-6'
6'	71.5	3.88, dd (4.8, 11.5)	C-6'	H-5'_a,b	C-5', C-7', C-8', C-10'	H-8, H-8', H-9', H-5'_a,b
7'	96.7	—	—	—	—	—
8'	71.0	3.61, d (3.2)	C-8'	H-9'	C-7'	H-6', H-9', H-8
9'	73.9	4.55, s	C-9'	H-8', H-10'	C-7', C-8', C-10'	H-6', H-8', H-10'
10'	83.3	4.39, s	C-10'	H-9'	C-6', C-8', C-9', C-11'	H-9'
11'	177.6	—	—	—	—	—

No.	δ_C	δ_H, Mult. (J in Hz)	HSQC	¹H-¹H COSY	HMBC	NOESY
2	154.8	7.84, s	C-2	—	C-4, C-6	H-2'
4	150.2	—	—	—	—	—
5	120.0	—	—	—	—	—
6	157.1	—	—	—	—	—
8	141.2	7.88, s	C-8	—	C-4, C-5	H-2', H-6', H-8'
1'	92.4	5.79, s	C-1'	H-2'	C-4, C-8, C-2', C-3'	H-2', H-3', H-4'
2'	82.1	4.53, s	C-2'	H-1', H-3'	C-1', C-3', C-4'	H-1', H-3', H-6'
3'	77.4	4.32, s	C-3'	H-2', H-4'	C-1', C-2', C-4'	H-1', H-2', H-4', H-5'_a,b
4'	81.2	4.62, s	C-4'	H-3', H-5'_a,b	C-3', C-6'	H-1', H-3', H-5'_a,b
5'_a,b	27.1	2.22~2.34, m	C-5'	H-4', H-6'	C-3', C-4', C-6', C-7'	H-3', H-4', H-6'
6'	71.5	3.88, dd (4.8, 11.5)	C-6'	H-5'_a,b	C-5', C-7', C-8', C-10'	H-8, H-8', H-9', H-5'_a,b
7'	96.7	—	—	—	—	—
8'	71.0	3.61, d (3.2)	C-8'	H-9'	C-7'	H-6', H-9', H-8
9'	73.9	4.55, s	C-9'	H-8', H-10'	C-7', C-8', C-10'	H-6', H-8', H-10'
10'	83.3	4.39, s	C-10'	H-9'	C-6', C-8', C-9', C-11'	H-9'
11'	177.6	—	—	—	—	—

No.	δ_C	δ_H, Mult. (J in Hz)	HSQC	¹H-¹H COSY	HMBC
2	144.6	8.40, s	C-2	—	C-4, C-6
4	148.1	—	—	—	—
5	118.1	—	—	—	—
6	149.9	—	—	—	—
8	143.7	8.75, s	C-8	—	C-4, C-5
1'	90.1	6.15, d (9.5)	C-1'	H-2'	C-4, C-8, C-2'
2'	80.2	4.65, s	C-2'	H-1', H-3'	C-3', C-4'
3'	75.4	4.38, s	C-3'	H-2', H-4'	C-1', C-4'
4'	78.7	4.67~4.69, m	C-4'	H-3', H-5'	C-6'
5'_a,b	24.8	2.33~2.21, m	C-5'	H-4', H-6'	C-3', C-4', C-6', C-7'
6'	66.7	4.39, dd (5.2, 11.8)	C-6'	H-5'_a,b	C-5', C-10'
7'	93.8	—	—	—	—
8'	90.9	—	—	—	—
9'	74.0	4.29, d (1.4)	C-9'	H-10'	C-7', C-8'
10'	77.7	4.56, d (1.4)	C-10'	H-9'	C-6', C-8', C-9', C-11'
11'	172.4	—	—	—	—

No.	δ_C	δ_H, Mult. (J in Hz)	HSQC	¹H-¹H COSY	HMBC
2	144.6	8.40, s	C-2	—	C-4, C-6
4	148.1	—	—	—	—
5	118.1	—	—	—	—
6	149.9	—	—	—	—
8	143.7	8.75, s	C-8	—	C-4, C-5
1'	90.1	6.15, d (9.5)	C-1'	H-2'	C-4, C-8, C-2'
2'	80.2	4.65, s	C-2'	H-1', H-3'	C-3', C-4'
3'	75.4	4.38, s	C-3'	H-2', H-4'	C-1', C-4'
4'	78.7	4.67~4.69, m	C-4'	H-3', H-5'	C-6'
5'_a,b	24.8	2.33~2.21, m	C-5'	H-4', H-6'	C-3', C-4', C-6', C-7'
6'	66.7	4.39, dd (5.2, 11.8)	C-6'	H-5'_a,b	C-5', C-10'
7'	93.8	—	—	—	—
8'	90.9	—	—	—	—
9'	74.0	4.29, d (1.4)	C-9'	H-10'	C-7', C-8'
10'	77.7	4.56, d (1.4)	C-10'	H-9'	C-6', C-8', C-9', C-11'
11'	172.4	—	—	—	—

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金核霉素生物合成途径的体内研究和关键代谢物鉴定

In vivo Studies on the Biosynthetic Pathway of Aureonuclemycin and Identification of Key Metabolites

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