海洋溶藻弧菌的培养优化及溶藻粉剂胶囊球工艺研究

发布时间：2018-01-02 14:25

本文关键词：海洋溶藻弧菌的培养优化及溶藻粉剂胶囊球工艺研究　出处：《深圳大学》2017年硕士论文　论文类型：学位论文

【摘要】：目前赤潮藻的防治主要以物理法、化学法为主,其防控效果总体达不到预期,并容易产生二次污染,因此利用生物手段,尤其是溶藻细菌防治有害赤潮藻的研究逐渐得到重视。国内外对溶藻细菌的研究主要集中在溶藻机制、活性物质提取、相关功能基因等研究,溶藻细菌固定化、溶藻活性物质粉剂化、胶囊化等应用工艺的研究相对较少,本论文将溶藻弧菌进行发酵优化,将发酵液上清经喷雾干燥获得溶藻粉剂,对粉剂进行包埋制备适应于实际赤潮防治的悬浮型胶囊球。本研究对溶藻弧菌H115(简称H115)进行溶藻特性的探究,确定藻菌比例为1:7000,作用10min作为单因素优化溶藻作用的条件;在溶藻方式探究实验中确定H115属于间接溶藻,且对数期后的H115溶藻物质才能大量累计,发挥较强的溶藻效果。本研究通过单因素法筛选出:碳源、氮源、温度和发酵时间这四个因子对H115溶藻活性物质的分泌影响较大;本研究采用U6(64)均匀设计表将上述四个因子进行均匀设计法优化,优化结果为:(1)H115溶藻效率最优组为:以2216 E为基础培养基,添加山梨醇:1.0%,peptone:2.0%,初始pH为7.5,发酵温度40℃,摇床转速200 rpm,发酵时间54 h;(2)H115的生长密度最优组合为:山梨醇添加比例为1.5%,peptone:3.0%,发酵温度24℃,发酵时间48h。本研究对优化条件进行5L、100L发酵罐的扩大试验,以藻菌个数比1:600添加,作用藻细胞4h时,5L发酵罐培养、100L发酵罐培养的溶藻效果分别达到:94.23%、97.08%,表明经优化后,H115可适用于大规模发酵,且溶藻能力得到提升。本研究采用喷雾干燥法,以H115发酵上清液为基础,按浓度梯度与佐料混合,制备五种不同比例的H115发酵上清粉剂:五种比例粉剂(50%、60%、70%、80%、90%)按一定比例作用藻液24h后的LD100分别为:0.33%、0.15%、0.13%、0.10%、0.10%;采用锐孔凝固浴法,研究制备了泡腾型的抑藻悬浮海藻酸钠胶囊球。经过单因素法优化胶囊球后,当胶囊球与血红哈卡藻比例为1:100时,其作用16h的溶藻效果为73.32%,在24h达到100%,经研究实验,所制胶囊球通过缓慢释放H115上清粉剂到藻液中,起到抑藻作用的同时能悬浮于液体表面,为赤潮的防治提供可能。
[Abstract]:The prevention and control of red tide algae by physical method, chemical method, the prevention and control of the overall effect is not up to expectations, and easy to produce two pollution, therefore the use of biological means, especially on prevention and control of harmful algae lysing bacteria gradually get attention. Studies on algicidal bacteria mainly concentrated in the algicidal mechanism active material, extraction, research related genes, algicidal bacteria immobilized algicidal substances, powder, capsule and application technology research is relatively few, this paper will Vibrioalginolyticus fermentation optimization, the fermentation supernatant obtained by spray drying of algicidal powder, suspension type capsule ball embedding system to adapt to the actual preparation of red tide prevention powder. The research of Vibrioalginolyticus H115 (H115) to explore the algicidal characteristics, determine the proportion of algae and bacteria 1:7000, 10min as the single factor optimization of algicidal effect in the condition; Study on the mode of H115 is an indirect algicidal algicidal experiment determined, and the logarithmic phase after H115 to a large number of algicidal substances accumulated, play a strong algicidal effect. This study screened by single factor method: carbon source, nitrogen source, temperature and fermentation time of these four factors affect the solubility of algicidal substances secretion this study uses U6 to H115; (64) uniform design table of the four factors of the uniform design method, the optimization results are as follows: (1) H115 algicidal efficiency optimal group: 2216 E as basic medium, adding sorbitol: 1%, peptone:2.0%, initial pH 7.5, fermentation temperature 40. Rotation speed of 200 RPM, fermentation time 54 h; (2) the optimal combination of growth density of H115 is: add sorbitol ratio of 1.5%, peptone:3.0%, fermentation temperature 24, fermentation time 48h. the study of 5L optimization conditions, expanding test of 100L fermentation tank, the number of algae bacteria than 1:600 added effect The algal cells 4h, 5L fermentation in 100L fermentor, the algae lysing effect respectively: 94.23%, 97.08%, show that after optimization, H115 can be applied to large scale fermentation, and the algae lysing ability has been improved. This study by spray drying method, using H115 based mixed fermentation supernatant, according to concentration gradient and seasoning H115, fermentation supernatant powder preparation of five different proportion: five ratio of powder (50%, 60%, 70%, 80%, 90%) according to a certain proportion of algae solution after 24h LD100 were 0.33%, 0.15%, 0.13%, 0.10%, 0.10%; the sharp hole bath method, effervescent type algal suspension study on the preparation of sodium alginate capsule ball. After optimization of capsule ball single factor method, when the ball capsule and blood red algae Haka ratio was 1:100, the effect of 16h on algicidal effect is 73.32%, reached 100% in 24h, the experimental results, the capsules prepared by the slow release of H115 ball to powder in the culture supernatant and to inhibition At the same time, the effect of algae can be suspended on the surface of the liquid, providing the possibility of the prevention and control of red tide.

【学位授予单位】：深圳大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：X55;TQ920.6

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