Oleic-acid-producing Bacillus-like Bacteria Screened by Means of Chemomics
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摘要: 利用高效液相色谱-四级杆飞行时间质谱联用技术测定了52个芽胞杆菌Bacillus-like菌株发酵上清液中的代谢物组成,各物质的分子特征通过MassHunter软件从原始数据中进行提取,然后通过Metlin代谢物质谱数据库的检索比对,获得各代谢物的基本信息。结果表明,在海绵芽胞杆菌Bacillus plakortidis菌株FJAT-14858的发酵上清液中,可以检测出游离油酸,其Metlin匹配得分达95.61,其相对含量占发酵上清液总代谢物的1.62%,精确质量数为284.2693。海绵芽胞杆菌菌株FJAT-14858是首个被发现产非毒性的游离油酸的芽胞杆菌菌株,本研究为新型微生物油脂的开发与利用提供菌种资源和理论依据。
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关键词:
- 海绵芽胞杆菌FJAT-14858 /
- 游离油酸 /
- 物质组学 /
- 高效液相色谱-四级杆飞行时间质谱
Abstract: Extracellular metabolites in the supernatants 52 Bacillus-like strains were analyzed by HPLC along with the quadrupole time-of-flight mass spectrometry (HPLC-Q-TOF-MS). Subsequently, the molecular feature extraction (MFE) was performed on the data using the software MassHunter followed by submitting the information for comparison against the mass spectrometry database, Metlin. Free oleic acid was thereby detected in the cultural supernatant of Bacillus plakortidis, FJAT-14858. Its matching score with oleic acid was 95.61, accurate mass was 284.2693, and relative content in the metabolites collected was up to 1.62%. To date, B. plakortidis FJAT-14858 was the first Bacillus-like bacterium known to produce free oleic acid with no cytotoxicity to the bacteria per se. -
表 1 供试芽胞杆菌菌株及产游离油酸的芽胞杆菌筛选结果
Table 1. Bacillus-like bacteria screened and FJAT-14858 that produced free oleic acid in liquid culture medium
序号 菌株编号 学名 中文名称 保留时间 匹配度 相对含量 精确分子量 质核比 1 FJAT-14205 Aneurinibacillus migulanus 米氏解硫胺素芽胞杆菌 0 2 FJAT-14856 Aneurinibacillus aneurinilyticus 解硫胺素硫胺素芽胞杆菌 0 3 FJAT-10042 Fictibacillus barbaricus 奇异虚构芽胞杆菌 0 4 FJAT-10043 Fictibacillus bataviensis 奇异虚构芽胞杆菌 0 5 FJAT-16152 Bacillus cereus 蜡样芽胞杆菌 0 6 FJAT-14221 Bacillus acidiceler 酸快生芽胞杆菌 0 7 FJAT-10032 Bacillus fordii 福氏芽胞杆菌 0 8 FJAT-10014 Bacillus alcalophilus 嗜碱芽胞杆菌 0 9 FJAT-14272 Bacillus cibi 食物芽胞杆菌 0 10 FJAT-14236 Bacillus butanolivorans 食丁酸芽胞杆菌 0 11 FJAT-14270 Bacillus ginsengihumi 人参土芽胞杆菌 0 12 FJAT-10033 Bacillus fortis 强壮芽胞杆菌 0 13 FJAT-520 Bacillus coagulaus 凝结芽胞杆菌 0 14 FJAT-10010 Bacillus endophyticus 内生芽胞杆菌 0 15 FJAT-14212 Bacillus indicus 印度芽胞杆菌 0 16 FJAT-14252 Bacillus isronensis 印空研芽胞杆菌 0 17 FJAT-8757 Bacillus badius 栗褐芽胞杆菌 0 18 FJAT-16342 Bacillus cereus 蜡样芽胞杆菌 0 19 FJAT-16366 Bacillus cereus 蜡样芽胞杆菌 0 20 FJAT-8762 Bacillus clausii 克劳芽胞杆菌 0 21 FJAT-14215 Bacillus cecembensis 科研中心芽胞杆菌 0 22 FJAT-16072 Bacillus megaterium 巨大芽胞杆菌 0 23 FJAT-10015 Alicyclobacillus cellulosilyticus 解纤维素脂环酸芽胞杆菌 0 24 FJAT-14213 Bacillus lehensis 列城芽胞杆菌 0 25 FJAT-17931 Bacillus amyloliquefaciens 解淀粉芽胞杆菌 0 26 FJAT-17934 Bacillus amyloliquefaciens 解淀粉芽胞杆菌 0 27 FJAT-2349 Bacillus amyloliquefaciens 解淀粉芽胞杆菌 0 28 FJAT-8754 Bacillus amyloliquefaciens 解淀粉芽胞杆菌 0 29 FJAT-10037 Bacillus hemicellulosilyticus 解半纤维素芽胞杆菌 0 30 FJAT-8764 Bacillus firmus 坚强芽胞杆菌 0 31 FJAT-16022 Bacillus methylotrophicus 甲基营养芽胞杆菌 0 32 FJAT-16041 Bacillus methylotrophicus 甲基营养芽胞杆菌 0 33 FJAT-8761 Bacillus circulans 环状芽胞杆菌 0 34 FJAT-14210 Bacillus koreensis 韩国芽胞杆菌 0 35 FJAT-16291 Bacillus licheniformis 地衣芽胞杆菌 0 36 FJAT-16398 Bacillus licheniformis 地衣芽胞杆菌 0 37 FJAT-16414 Bacillus licheniformis 地衣芽胞杆菌 0 38 FJAT-16766 Bacillus licheniformis 地衣芽胞杆菌 0 39 FJAT-16777 Bacillus licheniformis 地衣芽胞杆菌 0 40 FJAT-16973 Bacillus licheniformis 地衣芽胞杆菌 0 41 FJAT-17199 Bacillus licheniformis 地衣芽胞杆菌 0 42 FJAT-17228 Bacillus licheniformis 地衣芽胞杆菌 0 43 FJAT-17433 Bacillus licheniformis 地衣芽胞杆菌 0 44 FJAT-17730 Bacillus licheniformis 地衣芽胞杆菌 0 45 FJAT-8756 Bacillus azotoformans 产氮芽胞杆菌 0 46 FJAT-8765 Bacillus.flexus 弯曲芽胞杆菌 0 47 FJAT-14220 Bacillus aryabhattai 阿氏芽胞杆菌 0 48 FJAT-16042 Bacillus aryabhattai 阿氏芽胞杆菌 0 49 FJAT-16043 Bacillus aryabhattai 阿氏芽胞杆菌 0 50 FJAT-16059 Bacillus aryabhattai 阿氏芽胞杆菌 0 51 FJAT-14227 Bacillus novalis 休闲地芽胞杆菌 0 52 FJAT-14858 Bacillus plakortidis 海绵芽胞杆菌 45.6283 95.61 1.62 284.2693 283.2622 表 2 化合物鉴定的最佳匹配结果
Table 2. Best matching result for compound identification
Best Name Formula Score Mass Mass (DB) Diff (ppm) Diff (abs.ppm) Diff (mDa) Species m/z m/z (Calc) Diff (ppm) Diff (mDa) Height Height (Calc) Height /% Height /% (Calc) Hight Sum /% Hight Sum /% (Calc) Yes Oleic acid (D2) C18H32D2O2 95.61 284.2693 284.2684 -3.21 3.21 -0.91 (M-H)- 283.2622 283.2612 -3.77 -1.1 79558.5 77929.3 100 100 83.6 81.8 -
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