Microsoft Word 李梓铭-已校对.doc

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1 5 4 Vol. 5 No Journal of Food Safety and Quality Apr., , 李梓铭 2 1,, 谢靓 2 2,, 蒋立文 3*, 李云倩 (1., ; 2., ; 3., ) 1 摘要 : 目的 MRS, 方法 18% NaCl, PCR 16S rdna, 结果,, 结论 16S rdna 关键词 : ; ; Isolation and identification of salt-tolerance lactic acid bacterium from high-salt chili LI Zi-Ming 1, 2, XIE Jing 1, 2, JIANG Li-Wen 2, 3*, LI Yun-Qian 1 (1. College of Food Science and Technology, Hunan Agricultural University, Changsha , China; 2. Hunan Provincial Key Laboratory of Food Science and Biotechnology, Changsha , China; 3. Hunan Provincial Research Center of Engineering and Technology for Fermented Food, Changsha , China) ABSTRACT: Objective To isolate some salt-tolerance strains with different colony morphologies from Hunan characteristic fermented chilli samples using selected MRS medium. Methods One isolated strain which could grow on an 18% NaCl salt tolerant bacteria medium was identified base on the physiological and biochemical experiments with 16S rdna sequencing and phylogenetic tree analyses. Results This strain had a good ability of producing lactic acid and high tolerance to salt. Conclusion This strain was identified as Lactobacillus plantarum through 16S rdna sequence analysis. KEY WORDS: fermented chili; lactic acid bacteria; salt-tolerance 1 引言,,,,, NaCl, NaCl, NaCl 10%~15%,,, 基金项目 : (SCX1220) Fund: Supported by National Undergraduate Innovation Project (SCX1220) * 通讯作者 :,,, hnndjlw@163.com *Corresponding author: JIANG Li-Wen, Ph.D, Professor, Hunan Provincial Key Laboratory of Food Science and Biotechnology, Changsha , China. hnndjlw@163.com

2 4, : 1017 (Lactobacillus reuteri) / (Lactobacillus casei/paracasei) / (Lactobacillus paraplantarum/ pentosus) (Lactobacillus acidophilus) (Lactobacillus plantarum) (Lactobacillus rhamnosus) (Bifidobacterium ) [1], [2] ( ) [3,4],,,,,, [5,6],,, 2 材料与方法 2.1 实验材料材料 20% 试剂与仪器 : (95%),, ; : GZ-250-S ( ); PHS-3C ph ( ); 5417R ( Eppendorf); ( ); WS ( ) 培养基 MRS [7,8] ( ): 10 g 5 g 4 g 20 g -801 ml 2 g 5 g 2 g 0.2 g 0.05 g 15 g 2% 1000 ml ph 6; 10 g 5 g 4 g 20 g -801 ml 2 g 5 g 2 g 0.2 g 0.05 g 25 g 2% 10% 1000 ml ph 6; MC [9, 10] 2.2 试验方法耐盐菌株的分离 25 g, 225 ml, , 0.1 ml, 37 48~72 h,,, G(+), NaCl (5 % 10 % 12 % 15 % 18 % 20 %),, NaCl,, MRS, h, [11] 耐盐菌株的生理生化特性 (1) 0.1 ml, h,,,, (2), ~14 d,,, 10~20 min,, (3), 37 7~14 d, (4), 37

3 h, 0.2 ml,, 10~15 min (5),, (, B.C.P, ph 5.2, ph 6.8 ), (6) 5 ml, 10 % H2SO4 0.5 ml, 2% KMNO4 0.5 ml,,,,, (7), S rdna 的扩增与序列分析 DNA [12,13], 16S rdna, primer1 : AGAGTTTGATCCTGGCTCAG; primer2 : AAGGAGGTGATCCAGCCGCA, PCR : 95 5 min; s, s, s, min,, NCBI BLAST 3 结果与分析 3.1 菌株的分离筛选,, G(+) ly-1 Y-2 Y-3 ly-4 NaCl 5% 10 % 12 % 15 % 18 % 20 %,, ly-4, NaCl 18% ly-4,,,,,,, 3.2 菌株生理生化性质试验结果,,,,, 1 Fig. 1 Table 1 1 ly-4 ( 1000) Colonial morphology and thallus appearance of ly-4 ( 1000) 表 1 耐盐菌株生理生化特性 The salt-tolerance strains of physiological and biochemical characteristics : + ( ), ly-4 ( Lactobacillus ), (Lactobacillus ) [14]

4 4, : S rdna 序列鉴定 ly-4 16S rdna PCR 3, bp, 1500 bp 2 ly-4 PCR Fig. 2 Electrophoresis patterns of purified PCR products 2, 1%, 1500 bp,,, GenBank( KJ636841), NCBI BLAST, ly-4 16S rdna NCBI BLAST Lactobacillus plantarum WCFS1 strain WCFS1 99%, GenBank 16S rdna, MEGA5.02 Neighboringjoining 3 3, ly-4 Lactobacillus plantarum, ly-4 Lactobacillus planttarum 16S rdna 99% [14] 16S rdna, (Lactobacillus) (Lactobacillus plantarum) 4 讨论 ly-4, 18%NaCl, ly-4,,,, Fig. 3 3 ly-4 16S rdna Phylogenetic tree based on16s rdna gene sequences for ly-4 strain

5 参考文献 [1]. [J]., 2009(2): Zhang HP. Present situation and development countmeasures for probiotic lactic bacteria and fermented milk in China [J]. Dairy Sci Technol, 2009(2): [2] Lanyi JK. Salt-dependent properties of proteins from extremely halophilic bacteria [J]. Bacteriol Rev, 1974, 38(3): [3] Zahiroddini H,et al. Effect of microbial inoculants on the fermentation, nutrient retention and aerobic stability of barley silage [J]. Asian-Aust. J Anim. Sci. 2006(19): [4] Ammor MS,Mayo B. Selection criteria for lactic acid bacteria to be used as functional starter cultures in dry sausage production:an update [J]. Meat Sci, 2007, (01): [5],,,. [J]., 2010, 29 (05): Wang FJ, Lu F, Ju Y, et al. Separation and identification of lactic acid bacteria isolated from soy sauce mash [J]. China Brewing, 2010, 29 (05): [6],,. [J]., 2004(02): Huang CZ, Chen YR, Qi FL. Studied on the biological properties of Lactobacillus acidophilus [J]. Food Res Dev, 2004(02): [7],,. [J]., 2012, 24(8): Xin GQ, Xin GM, Xu HY. Isolation and identification of lactobacillus plantarum strain and studies on bio-character [J]. Acta Agric Jiangxi. 2012, 24(8): [8],,. [J]., 2011, 32(05): 1 4. Yu SJ, Peng YP, Deng YW. Research on the isolation and growth characteristics of acidithiobacillus ferrooxidan [J]. J Jiangxi Univ Sci Technol, 2011, 32(05): 1 4. [9],,. [J]., 2007(2): He L, Tang AJ, Yang GM. Study on fermentation mechanism of vegetable [J]. China Brewing. 2007(2): [10],,,. [J]., 2012, 28(1): Sha M, Pang HM, Gu LNZ, et al. Isolation and identification of lactic acid bacteria from natural fermentation peppers [J]. Food Mach, 2012, 28(1): [11],. [J]. ( ), 2011(11): Zhong YQ, Xia YB. Isolation and identification of lactic bacteria in natural fermentation pepper [J]. Acad Period Farm Products Process 2011(11):52 55 [12],,,. PCR-DGGE [J]., 2011(2): Liu F, Wang DX, Xu WM, et al. Bacterial diversity in water-boiled salted duck during storage analyzed by PCR-DGGE [J]. J Nanjing Agric Univ, 2011, 34(2): [13],,. (PCR) [J] , 27(5): Xu Y, Zhang LP, Gu GX. Novel advance on polymerase Chain reaction(pcr) for identification of spoilage bacteria in beer [J]. Food Ferment Ind, 2000, 27(5): [14],,,. [J]., 2007, 8(04): Shang J, Zhong FX, Wang YL, et al. Sepration and screening of lactic acid bacterium from several traditional fermented vegetables [J]. Food Sci, 2007, 28(04): 作者简介 ( 责任编辑 : 张宏梁 ) 李梓铭, 硕士研究生, 主要研究方向为食品生物技术 @qq.com 蒋立文, 教授, 博士, 主要研究方向为食品生物技术 hnndjlw@163.com

Microsoft Word 聂雪梅.doc

Microsoft Word 聂雪梅.doc 4 4 Vol. 4 No. 4 2013 8 Journal of Food Safety and Quality Aug., 2013 聂雪梅 *, 李立, 孙利, 高飞 (, 100123) 摘要 : 2011 12,, 2007~2011,,,, 关键词 : ; ; ; ; Trade and notification situation on food import and export