微生物學通報 FEB 20,2010,37(2):179～185
脫色希瓦氏菌(Shewanella decolorationis) S12還原不同電子受體的厭氧發(fā)酵罐培養(yǎng)方法
王博1,2,3,4,5 許玫英2,3,4 孫國萍2,3,4*
(1. 中國科學院華南植物園 廣東 廣州 510650)
(2. 廣東省微生物研究所 廣東 廣州 510070)
(3. 廣東省菌種保藏與應(yīng)用重點實驗室 廣東 廣州 510070)
(4. 廣東省微生物應(yīng)用新技術(shù)公共實驗室 廣東 廣州 510070)
(5. 中國科學院研究生院 北京 100039)
摘 要: 脫色希瓦氏菌Shewanella decolorationis S12在厭氧環(huán)境下能夠使用多種電子受體進行厭氧呼吸,。為了取得足夠的細胞量用于膜蛋白質(zhì)組學等科學研究的需要, 本研究選取無機小分子(硝酸鈉),、金屬離子(檸檬酸鐵)和有機大分子(偶氮染料莧菜紅)作為電子受體, 在使用確定成分的無機鹽培養(yǎng)基條件下, 使用不同濃度的電子供體和碳源對S12進行厭氧條件下靜置和發(fā)酵罐的優(yōu)化培養(yǎng), 采用連續(xù)補充電子受體的培養(yǎng)方式, 確認了電子供體和碳源的合適濃度, 建立了S12厭氧發(fā)酵罐培養(yǎng)方法,。相比傳統(tǒng)的靜置厭氧培養(yǎng), 厭氧發(fā)酵罐培養(yǎng)方法在保證了嚴格厭氧條件下高效率還原電子受體的同時, 還極大的提高了細胞生長密度,。連續(xù)補充電子受體的厭氧發(fā)酵罐培養(yǎng)的S12最大細胞密度最大分別可達到靜置厭氧培養(yǎng)細胞密度的325, 304, 369倍, 而生長時間也比靜置厭氧培養(yǎng)分別縮短了26.5%, 17.6%, 7.5%,。這為需要大量細胞和蛋白的細菌厭氧呼吸生長實驗建立了可行方法, 對于進行兼性厭氧呼吸的微生物的大規(guī)模厭氧培養(yǎng)具有借鑒意義,。
關(guān)鍵詞: 脫色希瓦氏菌S12, 厭氧發(fā)酵罐培養(yǎng), 電子受體, 細胞生長密度
The Method Study of Shewanella decolorationis S12 Reduce Different Electron Acceptors with Anaerobic Fermentor Culture
WANG Bo1,2,3,4,5 XU Mei-Ying2,3,4 SUN Guo-Ping2,3,4*
(1. South China Botanical Garden, Chinese Academy of Sciences, Guangzhou, Guangdong 510650, China)
(2. Guangdong Institute of Microbiology, Guangzhou, Guangdong 510070, China)
(3. Guangdong Provincial Key Laboratory of Microbial Culture Collection and Application, Guangzhou,
Guangdong 510070, China)
(4. Guangdong Open Laboratory of Applied Microbiology, Guangzhou, Guangdong 510070, China)
(5. Graduate School of the Chinese Academy of Sciences, Beijing 100039, China)
Abstract: Shewanella decolorationis S12 is a strain which can perform anaerobic respiration by using vari-ous electron acceptors under the anaerobic environment. In this study, to obtain the enough biomass and sat-isfy the need of scientific research such as membrane proteomics, we used the inorganic small molecule (so-dium nitrate), metal ion (ferric citrate) and organic macromolecule (azo dye amaranth) as sole terminal elec-tron acceptors, using different concentrations of electron donor and carbon source for anaerobic static culture and anaerobic fermentor culture of Shewanella decolorationis S12. The cells were cultured by fed-batch cul-tivation mode to confirm the optimal concentrations of electron donor and carbon source, finally the method of anaerobic fermentor culture for S12 was established. Comparing to the traditional anaerobic static culture method, anaerobic fermentor culture method not only ensured a high rate mass-transfer efficiency resulting in the effective reduction of electron acceptors, but also greatly increased the cell growth density, the max cell growth densities were increased to 325, 304, 369 times and cell growth times were decreased 26.5%, 17.6%, 7.5% separately. This method provided an effective way to culture a large number of cells and pro-tein under anaerobic respiration condition. The procedure described above would be significance for the studies which need biomass cultivation of Shewanella genus bacteria and other anaerobic microorganisms under fully controlled conditions.
Keywords: Shewanella decolorationis S12, Anaerobic fermentor culture, Electron acceptor, Cell growth density
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