Microsoft Word - 16傅丽君.doc

Size: px

Start display at page:

Download "Microsoft Word - 16傅丽君.doc"

衬屠
7 years ago
Views:

1 热带海洋学报 JOURNAL OF TROPICAL OCEANOGRAPHY 海洋生物学 2011 年第 30 卷第 2 期 : BS01 傅丽君 1,2, 安新丽 1,3, 李东 2, 许丽霞 2, 田蕴 1,3, 郑天凌 1., ; 2., ; ,3 : 1 BS01 Brevibacterium sp., Alexandrium tamarense BS01, 40% ph r min 1 48h, BS01 : 20g L 1 0.5g L 1 ph : ; ; ; ; 中图分类号 : X55 文献标识码 : A 文章编号 : (2011) Optimization of medium components and culture conditions of algicidal actinomycetes BS01 FU Li-jun 1,2, AN Xin-li 1,3, LI Dong 2, XU Li-xia 2, TIAN Yun 1,3, ZHENG Tian-ling 1,3 1. Key Laboratory of Ministry of Education for Coast and Wetland Ecosystems, School of Life Sciences, Xiamen University, Xiamen , China; 2. Department of Environment and Life Science, Putian University, Putian , China; 3. State Key Laboratory of Marine Environmental Science, Xiamen University, Xiamen , China Abstract: A strain of actinomycetes BS01 capable of lysing the toxic dinoflagellate Alexandrium tamarense was isolated from mangrove area of Zhangjiangkou in Fujian province. The medium components and cultural conditions for algicidal activity of actinomycete strain BS01 were optimized by single-factor and uniform tests in shake-flask. The initial ph, fermentation temperature, fermentation time, and inoculum were optimized through single-factor tests. Results showed that BS01 exhibited the best algicidal activity by using soluble starch as carbon source, sodium nitrate as nitrogen source, initial ph at 7.7, temperature at 27.2, and shaking at 150r min 1 in 100mL of 250mL conical flask for 48h. The optimal fermentation parameters of BS01 were: soluble starch 20g L 1, sodium nitrate 0.5g L 1, ph value 7.7, and temperature 27.2 by using uniform design experiments. Key words: algicidal actinomycete; Alexandrium tamarense; optimization of medium component; single-factor experiment; uniform design, [1],,,, [2 5], 收稿日期 : ; 修订日期 : 基金项目 : (2008AA09Z408); ( ); (CWe0902); (2010J01223); (JA10232) 作者简介 : (1975 ),,,,, 通信作者 : microzh@xmu.edu.cn

2 110 Vol. 30, No. 2 / Mar., 2011 [6 8] 2 [9], ;,,, [8], 50%,, [10 11] 1962, Saffeman [12], , % [13 18],,,,,, [19 20] BS01 Brevibacterium sp. ( ), BS01, 供试菌和供试藻 BS01 Brevibacterium sp., Alexandrium tamarense, 培养基 f/2 ; BS E 主要仪器 (PYX-300G-A, ); (SHP-250, ); (PHS-25, ); (DNM-9602G, ); (Sartorius BS224S, ); (Eppendorf, ); (ZHWY- 211B, ) 1.2 BS E, d 1% 100mL 2216E 250mL, r min 1 24h f/2, (20 1) 3500 lx, 12h 12h.. 100, R = A B A 100%,, R, A, B 发酵最佳碳源和氮源的筛选 2216E,,,,, r min -1 24h, 发酵最佳温度的确定 , 150r min 1 24h, 发酵最佳时间的确定 BS01, h BS 发酵最佳 ph 值的确定 ph , r min 1 24h, 发酵最佳溶氧量确定 250mL mL, r min 1 24h, 发酵最佳转速确定

3 : BS r min 1, 28 24h, 均匀设计优化发酵条件 U 6 (6 4 ) [21], ph 4,, 1 表 1 U 6 (6 4 ) 均匀设计因素水平表 Tab. 1 Factor and level in the uniform design /(g L 1 ) NaNO 3 /(g L 1 ) ph / BS 碳源氮源对 BS01 抑藻活性的影响 ;,, ;, ph,, BS01, 2216E, BS01, 72.57%, 30.91%( 1) BS01 BS01, BS01, 2216E,, 54.73%( 2) BS01 2 BS01 Fig. 2 Effects of different nitrogen sources on algicidal 1 BS01 Fig. 1 Effects of different carbon sources on algicidal 温度对 BS01 抑藻活性的影响 3, BS01, , 53.06% 44.06% 32.38%; 28, 24, 25.87%,,,, 28 BS 发酵时间对 BS01 抑藻活性的影响 4, BS01 2d

4 112 Vol. 30, No. 2 / Mar., BS01 Fig. 3 Effects of different temperatures on algicidal activity of BS01 5 ph BS01 Fig. 5 Effects of different ph on algicidal 4 BS01 Fig. 4 Effects of different fermentation time on algicidal, 59.30%, 48h 72h 96h 24h 120h, 48h BS ph 值对 BS01 抑藻活性的影响 5, ph BS01 ph , 49.91% 45.55%, ph BS01,, ph [22] ph 6 6.5,, 18.12% 20.34%, BS01 ph 7.5 BS 溶解氧对 BS01 抑藻活性的影响 6, 250mL mL, BS % 55.63% BS01 6 BS01 Fig. 6 Effects of medium volume per flask on algicidal 100mL,, l50ml 30.12% 100mL BS 转速对 BS01 抑藻活性的影响 7, 150r min 1 7 BS01 Fig. 7 Effects of shaking speed on algicidal activity of BS01

5 : BS BS01, 53.42%, r min 1, 150r min 1 BS [23],, [21] U 6 (6 4 ) BS01, 2, N =6 K =4 Uniform Design Version , 3 4, : = b 0 + b b b b 4 4 R=0.9995; F=262.1; 0.05; S=1.00e 3 ; R a =0.9976; 4, BS01 5, U i i, 2, 3, 表 2 均匀设计分析结果 Tab. 2 Results of UD U 6 (6 4 ) for optimization of cell growth /(g L 1 ) NaNO 3 /(g L 1 ) ph / /RFU x 1 x 2 x 3 x 4 y 表 3 变量方差分析表 Tab. 3 ANOVA of variable U=1.05e 3 K=4 U/K=2.62e -4 Q=1.00e 6 N-1-K=1 Q/(N-1-K)=1.00e -6 F=262.1 L=1.05e 3 N-1=5 表 4 残差分析表 Tab. 4 ANOVA of residual ( )/ e e e e e e e e e 表 5 回归系数对抑藻活性的影响 Tab. 5 Regression coefficient on the algicidal activity U i /% i x 1 U e 20 U 1 /U 4.01e e 7 x 2 U e 4 U 2 /U x 3 U e 4 U 3 /U x 4 U e 21 U 4 /U 2.01e e 8

6 114 Vol. 30, No. 2 / Mar., x 4,, e 15, (0.05,1,1) 161.4, 4 (0.05,1,1), ( ), Uniform Design Version 3.00 : 20g L 1 NaNO 3 0.5g L ph7.7 3 BS01, 4, 6, 6,,, BS01 ph NaNO 3 ; BS01 20g L 1 NaNO 3 0.5g L 1 ph BS01, [1] KRIENITZ L, BALLOT A, KOTUT K, et al. Contribution of hot spring cyanobacteria to the mysterious death of Lesser Flamingos at Lake Bogoria Kenya[J]. FEMS Microbiology Ecology, 2003, 43(2): [2] IMAMURA N, MOTOIKEL,SHIMADA N, et al. An efficient screening approach for anti-microcystis compounds based on knowledge of aquatic Microbial ecosystem[j]. J Antibiot, 2001, 54(6): [3],. [J]., 1996, 20(2): [4],,. [J]., 2010, 50(3): [5],,. [J]., 2010, 31(2): [6] WANG XIN, LI ZHIJIANG, SU JIANQIANG, et al. Lysis of a red-tide causing alga, Alexandrium tamarense, caused by bacteria from its phycosphere[j]. Biological Control, 2010, 52(2): [7],,,. [J]. :, 2006, 45( 2): [8],,. [J]., 2002, 15(5): [9] RASHIDAN K K, BIRD D F. Role of predatory bacteria in the termination of a cyanobacterial bloom[j]. Microb Ecol, 2001, 41(2): [10],,,. [J]., 2006, 33 (6): [11],. [J]., 2009, 31(6): [12] SAFFERMAN R S, MORRIS M E. Evaluation of natural products for algicidal properties[j]. Appl Environ Microbiol, 1962, 10(4): [13] WHYTE LG, MAULE A, CULLIMORE D R. Method for isolating cyanobacteria-lysing Streptomycetes from soil[j]. Journal of Applied Microbiology, 1985, 58(2): [14] BIABANI M A, BAAKE M, LOVISETTO B, et al. Anthranilamides: new antimicroalgal active substances from a marine Streptomycetes sp.[j]. J Antibiot, 1998, 51(3): [15] SIGEE D C, GLENN R, ANDREWS M J, et al. Biological control of cyanobacteria: principles and possibilities[j]. Hydrobiologia, 1999, 395/396: [16] CHOI H J. Streptomyces neyagawaensis as a control for the hazardous biomass of Microcystis aeruginosa (Cyanobacteria) in eutrophic freshwaters[j]. Biological Control, 2005, 3(2): [17],,,. [J]., 2009, 32(6): [18],,,. [J]., 2008, 28(11): [19],,,. W5 [J]., 2009, 32(2): [20],,,. AN02 [J]., 2007, 27(4): [21]. [M]. :, 1994: [22] 贇,,,. SG-001 [J]., 2009, 6(48): [23],. [J]., 2009, 8(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