微生物学通报 JUN 20, 2008, 35(6): 913~917 Microbiology 2008 by Institute of Microbiology, CAS 研究报告紫甘薯花色苷色素的抑菌作用研究 1 韩永斌 2 朱洪梅 1* 顾振新 1 范龚健

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1 微生物学通报 JUN 20, 2008, 35(6): 913~917 Microbiology 2008 by Institute of Microbiology, CAS 研究报告紫甘薯花色苷色素的抑菌作用研究 1 韩永斌 2 朱洪梅 1* 顾振新 1 范龚健 ( ) ( ) 摘要 : 本文研究了紫甘薯花色苷色素抑制大肠杆菌金黄色葡萄球菌啤酒酵母和黑曲霉的作用及机理结果表明 : 紫甘薯花色苷色素对大肠杆菌及金黄色葡萄球菌均有抑制作用, 并与其浓度呈正相关, 而对啤酒酵母和黑曲霉无抑制作用透射电镜观察和大肠杆菌生长曲线表明, 该色素的抑菌作用可能是通过增强细胞膜的通透性, 使细胞异常生长, 抑制对数生长期的细胞分裂, 使细胞质稀薄细胞解体 SDS-PAGE 分析表明, 紫甘薯花色苷对大肠杆菌蛋白表达影响不明显, 未见特征性条带的消失, 仅对部分蛋白质合成量有影响关键词 : 紫甘薯, 花色苷, 抑菌作用 Inhibitory Effect of Purple Sweet Potato Pigments on some Microorganisms HAN Yong-Bin 1 ZHU Hong-Mei 2 GU Zhen-Xin 1* FAN Gong-Jian 1 (1. Key Laboratory of Food Processing and Quality Control, Ministry of Agriculture, NAU, Nanjing ) (2. College of Life Sciences, Shanxi Normal University, Linfen ) Abstract: Inhibitory effect of purple sweet potato pigments (PSPPs) on E. coli, Staph. aureus, Sacc. cerevisiae and Asp. niger was investigated in this paper. The results showed that E. coli and Staph. aureus could be inhibited strongly by PSPPs and dependent on the concentration of PSPPs, but no inhibitive effect of PSPPs on Sacc.cerevisiae and Asp. niger was observed. Observation under Transmission Electron Microscope was carried out and growth curve of E. coli was ploted out. It was indicated that PSPPs might change the permeability of cell membrane, make cell excrescent and inhibit cell division. As a result, cytoplasm could become thin, disassembled and even die. SDS-PAGE analysis suggested that PSPA have some influence on some proteins synthesis, but no disappearance of special protein lines be found. Keywords: Purple sweet potato, Anthocyanins, Antibacterial effect (Ipomoes batats L.), [1 3], [4,5], [6] [7], 基金项目 :(No. BG ) * 通讯作者 :Tel: ; : guzx@ njau. edu. cn 收稿日期 : ; 接受日期 :

2 914 微生物学通报 2008, Vol.35, No.6, 1 材料与方法 1.1 材料紫甘薯 :,, 5 mm ~ 8 mm, 紫甘薯花色苷色素的制备 : (95 1 mol/l =9 1, V/V) 3 h, r/min 20 min, ( =1 2), AB-8 (0.01 HCl), 0.01 HCl, (40 ) ( mg/l) 供试菌种 :(E. coli) (Staph. aureus)(asp. niger) (Sacc. cerevisiae), 1.2 实验方法敏感性实验 : 20 ml,, 0.10 ml(10 5 CFU/mL), 0.05 ml, [8] h, h 最低抑菌浓度实验 : [9], (10 5 CFU/mL), h, (MIC) 抑制大肠杆菌实验 :, ml LB ( 0.5, 1, NaCl 0.5, ph 7.2), r/min, 0 h 2 h 4 h 6 h 8 h 10 h 12 h 24 h 36 h 48 h, OD 640, E. coli CK, 1 ml, AB 1/2 MIC 1 MIC; C D 4 h 1/2 MIC 1 MIC; EF 8 h 1/2 MIC 1 MIC SDS-PAGE, ml LB,, 1 ml 1 MIC, r/min, 4 h 8 h, 25 ml 5000 r/min 10 min, 2, 100 µl, 5 min~10 min, 5000 r/min 10 min, SDS-PAGE E. coli 5.0, 12 SDS-PAGE, 0.1 [10], 0.3, 4 15 min~30 min r/min~13000 r/min 10 min, 3, 1,, Epon812,,, H 结果与分析 2.1 紫甘薯花色苷抑菌活性 1 1,,,,,,,, 2.2 紫甘薯花色苷最小抑菌浓度 ( 2),,

: 915 表 1 紫甘薯花色苷对菌种抑菌活性 Table1 Antimicrobial activity of purple sweet potato anthocyanins on different microorganisms Index Antimicrobial circle diameter (mm) E. coli 11.13± 0.02 Staph. aureus 13.

3 紫甘薯花色苷对大肠杆菌生长的影响 2,, 表 2 紫甘薯花色苷最小抑菌浓度 (MIC) 的测定结果 Table 2 MIC of anthocyanins from purple sweet potato on different microorganisms Anthocyanin concentration (μg/ml) 800 400 200 100 50 25 12.5 6.

3 : 915 表 1 紫甘薯花色苷对菌种抑菌活性 Table1 Antimicrobial activity of purple sweet potato anthocyanins on different microorganisms Index Antimicrobial circle diameter (mm) E. coli 11.13± 0.02 Staph. aureus 13.93± 1.93 Sacc. cerevisiae Asp. niger 0 0 图 1 紫甘薯花色苷抑菌效果 Fig. 1 Antibacterial effect of anthocyanins from purple sweet potato 800 μg/ml 200 μg/ml, 2.3 紫甘薯花色苷对大肠杆菌生长的影响 2,, 表 2 紫甘薯花色苷最小抑菌浓度 (MIC) 的测定结果 Table 2 MIC of anthocyanins from purple sweet potato on different microorganisms Anthocyanin concentration (μg/ml) (E. coli) (Staph. aureus) (Sacc. cerevisiae) (Asp. niger) : ; +: Note: : No microbe growth; +: Microbe growth 图 2 紫甘薯花色苷对大肠杆菌生长曲线的影响 Fig. 2 Effect of anthocyanin from purple sweet potato on E. coli growth curve : CK: ; A C E: 0 h 4 h 8 h 1/2 MIC ; B D F: 0 h 4 h 8 h 1 MIC Note: CK: Control; A, C, E: 1/2 MIC anthocyanin from purple sweet potato was added at 0 h, 4 h and 8 h, respectively; B, D, F: 1 MIC anthocyanin from purple sweet potato was added at 0 h, 4 h and 8 h, respectively,,,,, E. coli,,, LB 2.4 紫甘薯花色苷对大肠杆菌抑菌率的影响 3,, ;,,

anthocyanin from purple sweet potato was added at 0 h, 4 h and 8 h, respectively 2.5 紫甘薯花色苷对大肠杆菌显微特征的影响 ( 4), E. coli,,,, 4 h,,,,,,,, 2.6 紫甘薯花色苷对大肠杆菌蛋白质合成的影响 SDS- ( 5),,, 60 kd 图 4 透射电镜观察花色苷对 E.

4 916 微生物学通报 2008, Vol.35, No.6,, 图 3 紫甘薯花色苷对大肠杆菌抑菌率的影响 Fig. 3 Inhibitory effect of anthocyanin from purple sweet potato on E. coli : CK: ; A C E: 0 h 4 h 8 h 1/2 MIC ; B D F: 0 h 4 h 8 h 1 MIC Note: CK: Control; A, C, E: 1/2 MIC anthocyanin from purple sweet potato was added at 0 h, 4 h and 8h, respectively; B, D, F: 1 MIC anthocyanin from purple sweet potato was added at 0 h, 4 h and 8 h, respectively 2.5 紫甘薯花色苷对大肠杆菌显微特征的影响 ( 4), E. coli,,,, 4 h,,,,,,,, 2.6 紫甘薯花色苷对大肠杆菌蛋白质合成的影响 SDS- ( 5),,, 60 kd 图 4 透射电镜观察花色苷对 E. coli 形态影响 Fig. 4 Effect of anthocyanin on morphology of E. coli by TEM : a: ( 39000); b: 1 MIC 4 h ( 33000); c, d: 1 MIC 8 h ( 20000) Note: E. coli cell were incubated in the absence of anthocyanins or in the presence of anthocyanins (1 MIC) for 4 h or 8 h, respectively. And the specimens were then prepared for transmission electron microscopy. a: Intact cell ( 39000); b: Cell treated for 4 h with 1 MIC anthocyanin ( 33000); c, d: Cell treated for 8 h with 1 MIC anthocyanin ( 20000) 3 讨论,, ph,, [11] LB ph 7.0,, ;,, E. coli Staph. aureus,,, [12], SDS-PAGE,,,,,,

: 917 batatas. Phytochemistry, 1992, 31: 2127 2130. [3] Teranara N, Kato Y, Nakamura M, et al. Six diacylated anthocyanins from storage roots of purple sweet potato, Ipomoea batatas.

5 : 917 batatas. Phytochemistry, 1992, 31: [3] Teranara N, Kato Y, Nakamura M, et al. Six diacylated anthocyanins from storage roots of purple sweet potato, Ipomoea batatas. Biosci Biotechnol Biochem, 1999, 63: [4] Furuta S, Suda I, Nishiba Y, et al. High tert-buthylperoxyl radical scavenging activities of potato cultivars with purple flesh. Food Sci Technol Int Tokyo, 1998, 4: [5] Suda I, Furuta S, Nishiba Y, et al. Reduction of liver induced by carbon tetrachloride in rats administered purple-colored sweet potato juice. Nippon Shokuhin Kagaku 图 5 E. coli SDS-PAGE 电泳图谱 Fig. 5 SDS-PAGE electrophoresis map of E. coli : CK: ; A B: 1 MIC 4 h 8 h; M: Note: CK: Control; A, B: 1 MIC anthocyanin treated for 4 h and 8 h respectively; M: Marker molecule protein,,,,, 参考文献 [1] Goda Y, Shimizu T, Kato Y. Two acylated anthocyanins from purple sweet potato. Phytochemistry, 1997, 44: [2] Odake K, Terahara N, Saito N, et al. Chemical structures of two anthocyanins from purple sweet potato, Ipomoea Kagaku Kaishi (in Japanese with English abstract), 1997, 44: [6] Cheng CL, Wang ZY. Bacteriostasic activity of Anthocyanin of Malva sylvestris. Journal of Forestry Research, 2006, 17(1): [7],.., 2005, 2: 47. [8],.., 1980, pp [9],,.., 1982, pp [10].., 1983, pp [11] Shi ZL, Bassa IA, Gabriel SL, et al. Anthocyanin pigments of sweet potato. Food Science, 1992, 57(3): [12],,.., 2004, 35(3):

: 307, [], [2],,,, [3] (Response Surface Methodology, RSA),,, [4,5] Design-Expert 6.0,,,, [6] VPJ33 ph 3,, ph, OD, Design-Expert 6.0 Box-Behnken, VPJ3

: 307, [], [2],,,, [3] (Response Surface Methodology, RSA),,, [4,5] Design-Expert 6.0,,,, [6] VPJ33 ph 3,, ph, OD, Design-Expert 6.0 Box-Behnken, VPJ3 微生物学通报 FEB 20, 2008, 35(2) 306~30 Microbiology 2008 by Institute of Microbiology, CAS tongbao@im.ac.cn 生物实验室响应面分析法优化副溶血性弧菌生长条件刘代新宁喜斌 * 张继伦 2 (. 200090) (2. 20202) 摘要

微生物学通报 JUN 20, 2008, 35(6): 913~917 Microbiology 2008 by Institute of Microbiology, CAS 研究报告 紫甘薯花色苷色素的抑菌作用研究 1 韩永斌 2 朱洪梅 1* 顾振新 1 范龚健

微生物学通报 JUN 20, 2008, 35(6): 913~917 Microbiology 2008 by Institute of Microbiology, CAS 研究报告紫甘薯花色苷色素的抑菌作用研究 1 韩永斌 2 朱洪梅 1* 顾振新 1 范龚健