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1 6 12 Vol. 6 No Journal of Food Safety and Quality Dec., 2015 / 朱育菁, 刘波 *, 郑梅霞, 陈峥 (, ) 摘要 : 目的 Bacillus niacini FJAT 方法 / (liquid chromatography-hybrid quadrupole time-of-flight mass spectrometry) FJAT MassHunter,, Metlin 结果 FJAT , Metlin 165,, 96.52, 5.31%, min, 结论 关键词 : : ; ; Analysis on phenyllactic acid from Bacillus niacini based on liquid chromatography-hybrid quadrupole time-of-flight mass spectrometry ZHU Yu-jing, LIU Bo *, ZHENG Mei-xia, CHEN Zheng (Agricultural Bio-resources Research Institute, Fujian Academy of Agricultural Sciences, Fuzhou , China) ABSTRACT: Objective To analyze and test on active substance from Bacillus niacini FJAT Methods Extracellular metabolites of Bacillus niacini FJAT were analyzed by liquid chromatographyhybrid quadrupole time-of-flight mass spectrometry. Molecular feature extraction (MFE) and database retrieval were applied by Agilent Mass Hunter software, for the information overall metabolites. Results Totally 811 compounds were detected, and 165 of them were identified by library searching of Metlin database. The match score of phenyllactic acid was and its relative content was 5.31%. The retention time of phenyllactic acid was about min, and its accurate mass was Conclusion The results provide the theory basis for development and utilization of phenyllactic acid from Bacillus niacini. KEY WORDS: liquid chromatography-hybrid quadrupole time-of-flight mass spectrometry; Bacillus niacini; extracellular metabolites; phenyllactic acid 基金项目 : ( )(2015R1018-1, 2015R ) Fund: Supported by National Natural Science Foundation of China ( ), Fujian Seeding Industry Innovation Project Fujian Resource Preservation Center of the Bacillus-like Bacteria, and the Commonweal Project of Fujian Province (2015R1018-1, 2015R ) * 通讯作者 :,, fzliubo@163.com *Corresponding author: LIU Bo, Professor, Fujian Academy of Agricultural Sciences, Fujian , China. fzliubo@163.com

2 引言 G+C,, [1] (Bacillus niacini) [2,3], [4], Kurata [5] ( , Phenyllactic acid, C 9 H 10 O 3, CAS : ), [6] [7] [8], [9,10] (Staphylococcus aureus) (Pseudomonas aeruginosa) [11], (Penicillium verrucosum) (Penicillium citrinum) (Penicillium roqueforti) [12],,, [13] [14],, (LC-QTOF-MS),,, 2 材料与方法 2.1 材料与试剂 : (Bacillus niacini FJAT-14202),, : NA ; TSB ( BD ) : ( JT Baker ), (CNW, ), ( JT Baker ) : Agilent 1260/6520( ); METTLER TOLEDO AL104( ); ZHWY-2102C( ); ph Sartorius PB-10( ) 2.2 实验方法 : FJAT-14202,, TSB, 30, 170 r/min 48 h :, r/min 10 min; (-20 ), 4 15 min, r/min 10 min,, 0.22 μm, 4 : Zorbax SB-Aq C 18 (2.1 mm 100 mm, 1.8 μm); : 30 ; : 2 μl; : A=10 mmol/l +, B= ; : 0.3 ml/min; : 0 min 5%B; 8 min 5%B; 38 min 95%B; 45 min 95%B; 47 min 5%B; 55 min 5%B; MS : ; : 350, : 5 L/min; : 30 psig; : 3500 V; : 100 V; 65.0 V; 100~3000 m/z MS : 3 结果与分析 3.1 烟酸芽胞杆菌 Bacillus niacini FJAT 发酵液 LC-MS 物质成分分析 FJAT , 811, Metlin 165, (Score) 80 18, 1 1% 7 : (1) (phenyllactic acid), 5.31%; (2)6- (6-hydroxy caproic acid), 4.14%; (3)PS(16:1(9Z)/0:0), 1.42%; (4)N,N- (N,N-diethylglycine), 1.29%; (5)threonolactone, 1.28%; (6) (fungichromin), 1.24% 3.2 烟酸芽胞杆菌 Bacillus niacini FJAT 发酵液中苯乳酸成分分析 FJAT , min,

3 12, : / , 5,,, [M-H] - [M+X] - (X ), [M-H] -, ,, Metlin,, 1 FJAT Fig. 1 TIC of Bacillus niacini FJAT Table 1 表 1 烟酸芽胞杆菌 Bacillus niacini FJAT 发酵液高匹配成分 High matching compositions of fermentation broth from Bacillus niacini FJAT No. m/z % CAS KEGG 1 phenyllactic acid C 9 H 10 O C hydroxy caproic acid C 6 H 12 O C PS(16:1(9Z)/0:0) C 22 H 42 N O 9 P N,N-diethylglycine C 6 H 13 NO C threonolactone C 4 H 6 O fungichromin C 35 H 58 O C (3-pyridyl)-2-hydroxytetrahydrofuran C 9 H 11 NO C a-fluoroallotetrahydrocortisol C 21 H 33 F O aculeacin A C 51 H 82 N 8 O C threonolactone C 4 H 6 O hydroxy caproic acid C 6 H 12 O C oleic acid(d2) C 18 H 32 D 2 O vasicinol C 11 H 12 N 2 O C Ala His His C 15 H 21 N 7 O phenyllactic acid C 9 H 10 O C alpha-hydroxy-isobutyric acid C 4 H 8 O ,4-dihydroxy-butanoic acid C 4 H 8 O (3S,5S)-3,5-diaminohexanoate C 6 H 14 N 2 O C01186

4 Fig. 2 2 FJAT Extracted compound chromatogram(ecc) of phenyllactic acid in Bacillus niacini FJAT Fig. 3 3 FJAT Extracted ion chromatogram (EIC) of phenyllactic acid in Bacillus niacini FJAT FJAT Fig. 4 Isotopic peak of phenyllactic acid in Bacillus niacini FJAT-14202

5 12, : / 4989 Fig. 5 5 FJAT Mass spectrum of phenyllactic acid in Bacillus niacini FJAT 讨论, ph,, [15] [16],, ph, ; [17] statine [18] [19] ;,, [20],, 5 结论,,, 5.31%, 96.52, 10.62% 参考文献 [1] Pandey RC, Guenther EC, Aszalos AA, Brajtburg J. Physicochemical and biological comparison of polyene macrolide antibiotics fungichromin, lagosin and cogomycin [J]. J Antibiot (Tokyo), 1982, 35(8): [2],,,. [J]., 2012, 51(6): Lu H, Wei SJ, Tu XQ, et al. Studies on antifugal activity and toxicity of new type polyene antibioticson four kinds of rice pathogenic fungi [J]. Hubei Agric Sci, 2012, 51(6): [3] Shih HD, Liu YC, Hsu FL, et al. Fungichromin:A substance from Streptomyces padanus with inhibitory effects on Rhizoctonia solani [J]. J Agric Food Chem, 2003, 51(1): [4],,,. 702 [J]., 2011, 33(5): Wei SJ, Zhang ZP, Tu XQ, et al. Studies on the isolation, identification and activity of anti-fusarium ocysporum secondary metabolites produced by Streptomyces sp. 702 [J]. Acta Agric Univ Jiangxi, 2011, 33(5): [5] Vilar VJP, Bothelho CMS, Boaventura RAR. Equilibrium and kinetic modeling of Cd biosorption by algae Gelidium and agar exraction algal waste [J]. Water Res, 2006, 40(2): [6],,,. [J]., 2015, 30(1): Liu B, Wang JP, Tao TS, et al. List of Species Mame for the Aacillus-like Bacteria [J]. Fujian J Agric Sci, 2015, 30(1): [7] Nagel M, Andreesen JR. Bacillus niacini sp. nov., a nicotinatemetabolizing mesophile isolated from soil [J]. Int J Syst Bacteriol, 1991, 41: [8] Ensign JC, Rittenberg SC. The pathway of nicotinic acid oxidation by a Bacillus species [J]. J Biol Chem, 1964, 239:

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