Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/91538
標題: 不同層狀雙氫氧化物(LDHs)之添加對於枯草桿菌 Bacillus subtilis CWS1在液態發酵生產表面素之探討
Effect of different layered double hydroxides addition on surfactin production by Bacillus subtilis CWS1 in the submerged culture
作者: Pei-Hsin Chang
章蓓昕
關鍵字: Bacillus subtilis
bio-surfactants
Layered Double Hydroxides
adsorption
枯草桿菌
生物界面活性劑
層狀雙氫氧化物
吸附
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摘要: 近年來,生物界面活性劑surfactin(從自然中生產之胜肽),被廣泛地運用在生物醫學上。雖然生物界面活性劑在生物醫學與植物病學上極具潛力,但量產及純化條件的資訊仍是有限的。先前本實驗室的結果指出以枯草桿菌Bacillus subtilis CWS1液態醱酵生產表面素surfactin,添加層狀雙氫氧化物Mg-Al-nitrate LDHs可以有效增加表面素之產量。 本研究將分為兩部分作探討,第一部分針對其吸附機制作探討,由XRD分析 LDHs層間距由吸附前的7.8 Å撐開至16.1 Å,藉此證實層狀雙氫氧化物會將表面素吸附其中,同時以HPLC分析LDHs之內含物,發現具有表面素之存在,並藉由SEM與EDX成分分析可觀察層狀雙氫氧化物吸附情形。 第二部分將針對不同比例之Mg-Al-nitrate LDHs以及Mg-Fe-nitrate LDHs添加對於枯草桿菌Bacillus subtilis CWS1發酵產表面素產量之探討,結果表示,添加6 g/L濃度的Mg-Al-nitrate LDHs主培養五天可得到表面素最大產量約3789 mg/L;添加6 g/L濃度的Mg-Fe-nitrate LHDs主培養第一天就產出4280 mg/L的表面素,四天即可產5130 mg/L的表面素,顯示用Mg-Fe-nitrate LDHs對於CWS1產表面素之效益更加突出,並針對菌量與表面素產量作與時間的關係圖,顯示菌體對於此添加物有很好的適應性,但是其產表面素能力依舊維持一動態平衡;經一系列實驗探討,顯示添加Mg-Fe-nitrate LDHs有助於表面素生產之改善,並具有工業應用之潛能。
Surfactin, one of the biological surfactants was found to possess great potential in biomedical and plant disease application. However, information about the production and purification condition is still limited. In the previous study, the addition of Mg-Al-nitrate LDHs in the liquid culture of Bacillus subtilis CWS1 could significantly increase the surfactin production This study includes two part. In the first part, XRD (X-ray diffractometer) was used to measure the basal spacing of LDHs. The surfactin intercalation involves an ionic exchange reaction of LDHs at 25℃ and under N2 atmosphere in the aqueous solution could yield a series of biomolecules-LDHs with basal spacing ranging from 7.8 Å of 16.1 Å, revealed by XRD analyses. The wide interlayer spacing might be attributed to the self-alignment of the biomolecules in the layer confinement, as analyzed the components by HPLC. In addition, SEM (scanning electron microscope) and EDX (Energy-dispersive X-ray spectrometry) was used to verify the adsorption of surfactin. The process of embedding biomolecules into the clay gallery provides a new method for synthesizing biomaterial/LDHs hybrids potentially useful in agriculture or biomedical design. In the second part, different LDHs, Mg-Al-nitrate LDHs and Mg-Fe-nitrate LDHs, were used as additive to the cultivation of Bacillus subtilis CWS1. In our study, 6 g/L of Mg-Al-nitrate LDHs addition gave the maximum surfactin production of 3789 mg/L after five days culture. However, 4280 mg/L of surfactin was produced in one day culture when 6 g/L of Mg-Fe-nitrate LHDs was added to the cultivation. The maximum production of 5130 mg/L was obtained in four days cultivation. These results indicated that addition of Mg-Fe-nitrate LHDs in Bacillus subtilis CWS1 culture could effectively enhance the production of surfactin. As seen from the time course of the cultuvation, we could found out Mg-Fe-nitrate LHDs addition was found harmless to Bacillus subtilis, the production of surfactin was kept in a dynamic equilibrium. This study provided an approach for improving surfactin production from Bacillus subtilis CWS1 via the addition of Mg-Fe-nitrate LDHs as a stimulator.
URI: http://hdl.handle.net/11455/91538
其他識別: U0005-2207201516364800
文章公開時間: 2018-07-28
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