基于单细胞拉曼光谱技术的葡萄球菌黄素生物合成分析

引用本文: 陶站华, 刘军贤, 师德强, 康健. 基于单细胞拉曼光谱技术的葡萄球菌黄素生物合成分析[J]. 分析化学, 2016, 44(3): 456-461. doi: 10.11895/j.issn.0253-3820.150729 shu

Citation: TAO Zhan-Hua, LIU Jun-Xian, SHI De-Qiang, KANG Jian. Analysis of Staphyloxanthin Biosynthesis Using Single Cell Raman Spectroscopy[J]. Chinese Journal of Analytical Chemistry, 2016, 44(3): 456-461. doi: 10.11895/j.issn.0253-3820.150729 shu

基于单细胞拉曼光谱技术的葡萄球菌黄素生物合成分析

陶站华 ^{1,
,} ,
刘军贤 ^2, ,
师德强 ^3, ,
康健 ^1,2,

1.
¹ 广西科学院生物物理实验室, 南宁 530003;
2.
² 广西师范大学物理科学与技术学院, 桂林 541004;
3.
³ 广西科学院国家非粮生物质能源工程技术研究中心, 南宁 530003

通讯作者: 陶站华

基金项目:
本文系广西自然科学基金(No.2014GXNSFAA118193) (No.2014GXNSFAA118193)

广西科学院基本科研业务费资助项目(No.15YJ22SL08)资助 (No.15YJ22SL08)

摘要: 利用光镊拉曼光谱技术研究吲哚对金葡菌细胞中葡萄球菌黄素合成的抑制作用以及色素含量在分批培养过程中的动态变化。收集经不同浓度吲哚(终浓度为0,0.2,0.6,0.8,1.2和1.5 mmol/L)处理后的以及不同培养时间的金葡菌单细胞的拉曼光谱,以光谱1523 cm^-1峰强度表征色素含量,并与紫外可见分光光度法得到的结果进行比较。结果表明,细菌拉曼光谱1523 cm^-1峰强度与分光光度法测得的色素含量有良好的线性关系,相关系数达0.9772;群体和单细胞水平的光谱数据均表明,吲哚可剂量依赖性地抑制葡萄球菌黄素的合成,色素含量降低幅度超过70%;在分批培养中细菌色素含量在对数生长中期(12 h)达到最大值,各个时间点的群体内部细胞间色素含量的异质性较小,RSD在39.2%~61.1%之间。本研究表明光镊拉曼光谱技术是一种在单细胞水平分析葡萄球菌黄素含量的可靠方法。

关键词:

English

Analysis of Staphyloxanthin Biosynthesis Using Single Cell Raman Spectroscopy

1.
¹ Lab of Biophysics, Guangxi Academy of Sciences, Nanning 530003, China;
2.
² College of Physics and Technology, Guangxi Normal University, Guilin 541004, China;
3.
³ National Engineering Research Center for Non-Food Biorefinery, Guangxi Academy of Sciences, Nanning 530003, China

Corresponding author: 陶站华

Received Date: 15 September 2015
Fund Project:
本文系广西自然科学基金(No.2014GXNSFAA118193)

广西科学院基本科研业务费资助项目(No.15YJ22SL08)资助

Abstract: Laser tweezers Raman spectroscopy (LTRS) was used to study the inhibitory effect of indol on staphyloxanthin biosynthesis in Staphylococcus aureus cells and the dynamic changes of this pigment content inside bacterial cells during batch cultivation. The Raman spectra of Staphylococcus aureus cells cultivated for different time and exposed to various doses of indol were acquired. The intensity of 1523 cm^-1 band was used for the quantification of staphyloxanthin, in the meantime, the pigment was measured by UV spectrometry. The experimental result showed that an excellent linear relationship existed between the intensities of Raman peak at 1523 cm^-1 of bacterial cells and the pigment contents estimated by UV spectrometry, with a correlation coefficient of 0.9772. The spectral data at population level as well as single cell level revealed that indol could inhibit the production of pigment in dose-dependent manner, and the pigment content in bacterial cells incubated with indol decreased by 70%. Under the batch growth condition, the pigment amount in Staphylococcus aureus cells reached the maximum value during the middle exponential growth phase (12 h) and the heterogeneity of pigment content in bacterial cells within certain populations at various time points was relatively small, with RSDs of 39.2% to 61.1%. This investigation indicates that LTRS can be served as a reliable method for the analysis of staphyloxanthin content at single cell level.

Key words:

1. [1]
  1 Lindström S. Single Cell Analysis. Stockholm: Humana Press, 2012: 1-121 Lindström S. Single Cell Analysis. Stockholm: Humana Press, 2012: 1-12
2. [2]
  2 AI Min, LIU Jun-Xian, HUANG Shu-Shi, WANG Gui-Wen, CHEN Xiu-Li, CHEN Zhi-Cheng, YAO Hui-lu. Chinese J. Anal. Chem., 2009, 37(5): 758-763艾敏, 刘军贤, 黄庶识, 王桂文, 陈秀丽, 陈植成, 姚辉璐. 分析化学, 2009, 37(5): 758-7632 AI Min, LIU Jun-Xian, HUANG Shu-Shi, WANG Gui-Wen, CHEN Xiu-Li, CHEN Zhi-Cheng, YAO Hui-lu. Chinese J. Anal. Chem., 2009, 37(5): 758-763艾敏, 刘军贤, 黄庶识, 王桂文, 陈秀丽, 陈植成, 姚辉璐. 分析化学, 2009, 37(5): 758-763
3. [3]
  3 LAI Jun-Zhuo, LIU Bin, WANG Gui-Wen, TAO Zhan-Hua, HUANG Shu-Shi. Spectroscopy and Spectral Analysis, 2011, 31(2): 412-417赖钧灼, 刘斌, 王桂文, 陶站华, 黄庶识. 光谱学与光谱分析, 2011, 31(2): 412-4173 LAI Jun-Zhuo, LIU Bin, WANG Gui-Wen, TAO Zhan-Hua, HUANG Shu-Shi. Spectroscopy and Spectral Analysis, 2011, 31(2): 412-417赖钧灼, 刘斌, 王桂文, 陶站华, 黄庶识. 光谱学与光谱分析, 2011, 31(2): 412-417
4. [4]
  4 Xie C, Dinno M A, Li Y Q. Optics Letters, 2002, 27(4): 249-2514 Xie C, Dinno M A, Li Y Q. Optics Letters, 2002, 27(4): 249-251
5. [5]
  5 WANG Yan-Jun, YAO Hui-Lu, WANG Gui-Wen, WANG Yun, FENG Mei-Fu. Spectroscopy and Spectral Analysis, 2009, 29(7): 1881-1883王雁军, 姚辉璐, 王桂文, 汪蕴, 丰美福. 光谱学与光谱分析, 2009, 29(7): 1881-18835 WANG Yan-Jun, YAO Hui-Lu, WANG Gui-Wen, WANG Yun, FENG Mei-Fu. Spectroscopy and Spectral Analysis, 2009, 29(7): 1881-1883王雁军, 姚辉璐, 王桂文, 汪蕴, 丰美福. 光谱学与光谱分析, 2009, 29(7): 1881-1883
6. [6]
  6 Xie C G, Li Y Q. J. Appl. Phys., 2003, 93(5): 2982-29866 Xie C G, Li Y Q. J. Appl. Phys., 2003, 93(5): 2982-2986
7. [7]
  7 ZHOU Bing, LU Ming-Qian, ZHAO Li-Wei, HUANG Shu-Shi, CHEN Li-Mei. Chinese J. Anal. Chem., 2013, 41(12): 1789-1794周冰, 卢明倩, 赵丽伟, 黄庶识, 陈丽梅. 分析化学, 2013, 41(12): 1789-17947 ZHOU Bing, LU Ming-Qian, ZHAO Li-Wei, HUANG Shu-Shi, CHEN Li-Mei. Chinese J. Anal. Chem., 2013, 41(12): 1789-1794周冰, 卢明倩, 赵丽伟, 黄庶识, 陈丽梅. 分析化学, 2013, 41(12): 1789-1794
8. [8]
  8 Tang H, Yao H, Wang G, Wang Y, Li Y Q, Feng M. Optics Express, 2007, 15(20): 12708-127168 Tang H, Yao H, Wang G, Wang Y, Li Y Q, Feng M. Optics Express, 2007, 15(20): 12708-12716
9. [9]
  9 Tao Z H, Wang G W, Xu X D, Yuan Y F, Wang X, Li Y Q. FEMS Microbiol. Lett., 2011, 314(1): 42-489 Tao Z H, Wang G W, Xu X D, Yuan Y F, Wang X, Li Y Q. FEMS Microbiol. Lett., 2011, 314(1): 42-48
10. [10]
  10 Shao J W, Lin M M, Li Y Q, Li X, Liu J X, Liang J P, Yao H L. Plos One, 2012, 7(10):10 Shao J W, Lin M M, Li Y Q, Li X, Liu J X, Liang J P, Yao H L. Plos One, 2012, 7(10):
11. [11]
  11 Shao J W, Yao H L, Meng L J, Li Y Q, Lin M M, Li X, Liu J X, Liang J P. Vib Spectrosc, 2012, 63367-6337011 Shao J W, Yao H L, Meng L J, Li Y Q, Lin M M, Li X, Liu J X, Liang J P. Vib Spectrosc, 2012, 63367-63370
12. [12]
  12 Fitzgerald-Hughes D, Devocelle M, Humphreys H. FEMS Immunol. Med. Microbiol., 2012, 65(3): 399-41212 Fitzgerald-Hughes D, Devocelle M, Humphreys H. FEMS Immunol. Med. Microbiol., 2012, 65(3): 399-412
13. [13]
  13 Clauditz A, Resch A, Wieland K P, Peschel A, Gotz F. Infect. Immun., 2006, 74(8): 4950-495313 Clauditz A, Resch A, Wieland K P, Peschel A, Gotz F. Infect. Immun., 2006, 74(8): 4950-4953
14. [14]
  14 Daum R S. New England J. Med., 2008, 359(1): 85-8714 Daum R S. New England J. Med., 2008, 359(1): 85-87
15. [15]
  15 Oldfield E. Accounts Chem. Res., 2010, 43(9): 1216-122615 Oldfield E. Accounts Chem. Res., 2010, 43(9): 1216-1226
16. [16]
  16 Lee J H, Park J H, Cho M H, Lee J. Current Microbiology, 2012, 65(6): 726-73216 Lee J H, Park J H, Cho M H, Lee J. Current Microbiology, 2012, 65(6): 726-732
17. [17]
  17 Kocher S, Breitenbach J, Muller V, Sandmann G. Arch. Microbiol., 2009, 191(2): 95-10417 Kocher S, Breitenbach J, Muller V, Sandmann G. Arch. Microbiol., 2009, 191(2): 95-104
18. [18]
  18 Vitek P, Jehlicka J, Edwards H G, Osterrothova K. Anal. Bioanal. Chem., 2009, 393(8): 1967-197518 Vitek P, Jehlicka J, Edwards H G, Osterrothova K. Anal. Bioanal. Chem., 2009, 393(8): 1967-1975
19. [19]
  19 Hintze J L, Nelson R D. Am. Stat, 1998, 52(2): 181-18419 Hintze J L, Nelson R D. Am. Stat, 1998, 52(2): 181-184

扫一扫看文章

计量

PDF下载量: 0
文章访问数: 433
HTML全文浏览量: 28

通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

基于单细胞拉曼光谱技术的葡萄球菌黄素生物合成分析

通讯作者: 陶站华

English

Analysis of Staphyloxanthin Biosynthesis Using Single Cell Raman Spectroscopy

Corresponding author: 陶站华

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南：你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

计量

通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

基于单细胞拉曼光谱技术的葡萄球菌黄素生物合成分析

通讯作者: 陶站华

English

Analysis of Staphyloxanthin Biosynthesis Using Single Cell Raman Spectroscopy

Corresponding author: 陶站华

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南： 你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

计量

文章相关

通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

CCS Chemistry：颜徐州、鲍哲南：你的皮肤可能是一片SPMs