Formation and Determination of Saquinavir from Liquid Fermentation of Bacillus aryabhattai FJAT-14220
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摘要: 对阿氏芽胞杆菌Bacillus aryabhattai FJAT-14220发酵液中沙奎那维成分进行分析与检测。采用高效液相四级杆飞行时间质谱（liquid chromatography-hybrid quadrupole time-of-flight mass spectrometry）分析阿氏芽胞杆菌FJAT-14220发酵液中胞外代谢物的成分。为获得代谢物的信息，首先采用MassHunter软件对原始数据进行分子特征提取，再通过Metlin数据库检索比对。在阿氏芽胞杆菌FJAT-14220发酵液中检测到883种代谢物，通过Metlin谱库搜索获得初步鉴定的有148种。其中，发现了具有生物活性的物质沙奎那维Saquinavir，其匹配得分达到93.08，占发酵液总代谢物相对含量的1.73%，保留时间为2.719 1 min，精确质量数为670.383 7。沙奎那维的发现为阿氏芽胞杆菌来源的多醚类抗生素的开发与利用提供理论依据。
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关键词:
- 液相四级杆飞行时间质谱 /
- 阿氏芽胞杆菌 /
- 胞外代谢物 /
- 沙奎那维
Abstract: Extracellular metabolites of Bacillus aryabhattai FJAT-14220 during fermentation were analyzed with liquid chromatography-hybrid quadrupole time-of-flight mass spectrometry. Saquinavir in the fermentation broth was targeted for the study. Molecular feature extraction and database retrieval were applied to access the information on the metabolites of B. aryabhattai. A total of 883 compounds were detected with 148 of them identified. The matching score of saquinavir was 93.08 with a relative concentration of 1.73%. Its retention time was 2.7191 min; and, accurate mass, 670.3837. The results could be the foundation for future development and utilization of saquinavir by B. aryabhattai fermentation. -
图 1 阿氏芽胞杆菌B. aryabhattai FJAT-14220发酵液的LC-MS总离子流
Figure 1. TIC of LC-MS of B. aryabhattai FJAT-14220 fermentation broth
图 2 阿氏芽胞杆菌FJAT-14220发酵液中沙奎那维化合物提取色谱
Figure 2. Chemical chromatogram of extracted saquinavir in B. aryabhattai FJAT-14220 fermentation broth
图 3 阿氏芽胞杆菌FJAT-14220发酵液中沙奎那维离子提取色谱
Figure 3. Ion chromatogram of extracted saquinavir in B. aryabhattai FJAT-14220 fermentation broth
图 4 阿氏芽胞杆菌FJAT-14220发酵液中沙奎那维同位素峰结果
Figure 4. Isotopic peak of extracted saquinavir in B. aryabhattai FJAT-14220 fermentation broth
图 5 阿氏芽胞杆菌FJAT-14220发酵液中沙奎那维质谱
Figure 5. Mass spectrum of extracted saquinavir in B. aryabhattai FJAT-14220 fermentation broth
表 1 阿氏芽胞杆菌B. aryabhattai FJAT-14220发酵液中匹配得分高的成分
Table 1. Composition of extracts from B. aryabhattai FJAT-14220 fermentation broth with high matching scores
序号 化合物名称 分子式 匹配得分 分子量 质荷比 保留时间/min 相对含量 /% CAS 号 KEGG 号 1 6-hydroxy caproic acid C6 H12 O3 99.07 132.0785 131.0712 2.0668 0.45 - C06103 2 N,N-Diethylglycine C6H13 N O2 98.92 131.0944 130.0872 1.4177 1.03 - C16647 3 Ganglioside GD2 (d18:1/14:0) C75 H131 N3O34 98.18 1617.8607 1616.851 32.4205 0.50 - - 4 Oleic acid(d2) C18 H32D2 O2 97.27 284.2678 283.2606 45.6347 0.69 - - 5 Saquinavir C38 H50 N6 O5 93.08 670.3837 669.3767 2.7191 1.73 127779-20-8 C07243 6 Threonolactone C4H6 O4 87.35 118.0267 117.0194 1.442 0.46 21730-93-8 - 7 4-Oxoproline C5H7 N O3 87.16 129.0423 128.035 1.3205 1.11 - C01877 8 2,4-dihydroxy-butanoic acid C4H8 O4 85.99 120.0422 119.035 1.3204 0.09 - - 9 Hedamycin C41H50 N2O11 85.70 746.3382 745.3309 3.3364 0.31 11048-97-8 C15759 10 Arbekacin C22 H44 N6O10 85.67 552.3102 551.303 2.0189 0.54 51025-85-5 - 11 N-dodecanoyl-L-Homoserine lactone-3-hydrazone-biotin C26 H43N5 O5 S 84.26 537.2984 536.2911 2.7215 0.07 - - 12 Cypendazole C16 H19 N5 O3 81.79 329.1493 328.1425 1.4949 0.08 28559-00-4 C18917 13 2alpha-Fluoro-17beta-hydroxyandrost-4-en-3-one C19H27 F O2 81.52 306.1989 341.1675 40.4596 0.04 - C15276 
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