本文選題：細(xì)胞形態(tài) + 簡單節(jié)桿菌　； 參考：《天津科技大學(xué)》2012年碩士論文

【摘要】：微生物的有機(jī)溶劑耐性機(jī)制非常復(fù)雜,具有明顯的菌種特異性和溶劑選擇性,本論文以甾體C1,2脫氫反應(yīng)菌種-簡單節(jié)桿菌(Arthrobacter simplex)為研究模型,系統(tǒng)研究了有機(jī)溶劑對(duì)微生物細(xì)胞形態(tài)的影響。 首先,通過傳統(tǒng)紫外誘變結(jié)合高濃度乙醇馴化的方法,選育得到乙醇耐受性突變株TCCC11037-UV15X1-2,該菌株在乙醇濃度10%(v/v)時(shí)仍表現(xiàn)出一定的生長活性,并具有良好的遺傳穩(wěn)定性。出發(fā)菌株和耐性突變株均能利用乙醇作為唯一碳源生長,且耐性突變株表現(xiàn)出更高的乙醇利用能力,乙醇濃度為6%時(shí)菌體仍表現(xiàn)出良好的生長。 乙醇培養(yǎng)初期添加對(duì)耐性突變株和出發(fā)菌株生長均產(chǎn)生了一定的抑制作用,且對(duì)出發(fā)菌株的抑制作用更為顯著。隨著乙醇濃度的增加,最大細(xì)胞濃度和比生長速率均顯著降低,乙醇濃度為6%時(shí)分別為對(duì)照組的51.86%和21.90%,耐性突變株在乙醇添加濃度為2%時(shí)表現(xiàn)出的生長特性,最大細(xì)胞濃度和比生長速率分別0.822和0.106h-1,與未添加乙醇的對(duì)照組的0.732和0.108h-1相比幾乎相同。當(dāng)乙醇濃度為8%時(shí)耐性突變株延滯期延長,但之后又顯示出一定的生長；乙醇對(duì)數(shù)中后期添加時(shí),出發(fā)菌株的生長明顯受到抑制,并隨乙醇濃度的增加抑制作用逐漸增強(qiáng)。對(duì)于耐性突變株,乙醇濃度在2%-4%范圍內(nèi),菌體生物量均呈現(xiàn)較明顯的增長趨勢(shì)。與出發(fā)菌株相比,耐性突變株具有一定的乙醇耐受性,且在4%濃度范圍內(nèi)乙醇對(duì)耐性突變株的生長存在促進(jìn)作用。 對(duì)于出發(fā)菌株,對(duì)數(shù)中后期添加不同濃度的乙醇,在乙醇濃度0-6%的的范圍內(nèi),細(xì)胞的比表面積隨著乙醇濃度的增加不斷減小,最小減至8.334(對(duì)照組為10.527),并表現(xiàn)出明顯的劑量依賴性。當(dāng)乙醇?jí)毫σ瞥?菌體的細(xì)胞形態(tài)能夠可逆回復(fù)。說明細(xì)胞形態(tài)通過降低與有機(jī)溶劑的接觸面積而表現(xiàn)出一定的適應(yīng)性,這種變化由乙醇誘導(dǎo)產(chǎn)生。而對(duì)于耐性突變株,未添加乙醇時(shí)細(xì)胞的比表面積為8.661,明顯小于出發(fā)菌株(10.527),說明耐性菌株通過減小其比表面積降低與溶劑的接觸面積,抵御乙醇對(duì)菌體的不利影響。并且其細(xì)胞比表面積不會(huì)隨著乙醇濃度的改變和乙醇的撤除發(fā)生明顯變化,說明耐性突變的細(xì)胞形態(tài)已經(jīng)發(fā)生了一種較為穩(wěn)定、長期的變化。靜息體系下乙醇對(duì)細(xì)胞形態(tài)的研究表明,細(xì)胞形態(tài)對(duì)有機(jī)溶劑作出的響應(yīng)能夠在較短時(shí)間內(nèi)完成,乙醇處理3h和24h后細(xì)胞表面積和比表面積沒有明顯波動(dòng)。此外,靜息體系下細(xì)胞形態(tài)的變化與有機(jī)溶劑的濃度沒有明顯的劑量依賴關(guān)系。
[Abstract]:The mechanism of organic solvent tolerance of microbes is very complex, with obvious strain specificity and solvent selectivity. In this paper, Arthrobacter simplexer, a steroidal C _ 1O _ 2 dehydrogenation reaction strain, was used as the research model.The effects of organic solvents on the morphology of microorganism were systematically studied.Firstly, the ethanol tolerant mutant TCCC11037-UV15X1-2 was obtained by traditional ultraviolet mutagenesis combined with high concentration ethanol acclimation. TCCC11037-UV15X1-2 still showed certain growth activity and good genetic stability when ethanol concentration was 10g / v.Both the original strain and the tolerant mutant could grow with ethanol as the sole carbon source, and the tolerant mutant showed higher ethanol utilization ability, and the bacteria still showed good growth when ethanol concentration was 6.In the early stage of ethanol culture, the growth of resistant mutants and parent strains was inhibited by adding ethanol, and the inhibitory effect on the original strain was more significant.With the increase of ethanol concentration, the maximum cell concentration and specific growth rate decreased significantly. When ethanol concentration was 6, it was 51.86% and 21.90% of the control group, respectively.The maximum cell concentration and specific growth rate were 0.822 and 0.106 h-1 respectively, which were almost the same as 0.732 and 0.108h-1 in the control group without ethanol.When the ethanol concentration was 8, the delay period of tolerant mutant was prolonged, but the growth of the mutant was shown to be certain. When the ethanol concentration was added in the middle and late period of ethanol, the growth of the original strain was obviously inhibited, and the inhibitory effect was gradually enhanced with the increase of ethanol concentration.For tolerant mutants, the biomass of bacteria increased significantly in the range of 2% -4% ethanol concentration.Compared with the original strain, the tolerant mutant had certain ethanol tolerance, and ethanol promoted the growth of the tolerant mutant in the range of 4%.For the original strain, the specific surface area of the cells decreased with the increase of ethanol concentration in the range of 0-6% ethanol concentration, when different concentrations of ethanol were added in the late logarithmic period.The minimum was reduced to 8.334 (the control group was 10.527) and showed a significant dose dependence.When the ethanol pressure was removed, the cell morphology was reversible.The results showed that the cell morphology showed adaptability by decreasing the contact area with organic solvents, which was induced by ethanol.For tolerant mutants, the specific surface area of the cells without ethanol was 8.661, which was significantly smaller than that of the original strain (10.527), which indicated that the resistant strain could resist the adverse effect of ethanol on the bacteria by reducing its specific surface area and reducing the contact area with the solvent.The specific surface area of the cells did not change obviously with the change of ethanol concentration and ethanol removal, which indicated that the cell morphology of tolerant mutation had been changed steadily and for a long time.The study of ethanol on cell morphology in resting system showed that the response of cell morphology to organic solvent could be completed in a relatively short time, and there was no obvious fluctuation of cell surface area and specific surface area after ethanol treatment for 3 h and 24 h.In addition, there was no significant dose-dependent relationship between the changes of cell morphology and the concentration of organic solvents in the resting system.
【學(xué)位授予單位】：天津科技大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R378

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 龔萬本;關(guān)怡新;王海清;姚善涇;;簡單節(jié)桿菌催化6α-甲基醋酸可的松C_(1,2)脫氫動(dòng)力學(xué)[J];高�；瘜W(xué)工程學(xué)報(bào);2007年06期

2 葉麗,史濟(jì)平;甾體微生物轉(zhuǎn)化在制藥工業(yè)中的應(yīng)用[J];工業(yè)微生物;2001年04期

3 李曉靜,陽葵,馮霞;絲狀微生物形態(tài)分析方法及影響形態(tài)的因素[J];化學(xué)工業(yè)與工程;2002年03期

4 張金紅,張秀明,陳長治,李寶安,俞耀庭;有機(jī)相中梨頭霉菌催化11-脫氧皮質(zhì)醇轉(zhuǎn)化成氫化可的松的研究[J];南開大學(xué)學(xué)報(bào)(自然科學(xué)版);1998年04期

5 徐詩偉,徐清,法幼華;甾體1,4-脫氫和11α-羥基化反應(yīng)的兩種不同微生物轉(zhuǎn)化[J];生物工程學(xué)報(bào);2000年05期

6 徐詩偉,徐清,曹桂芳,法幼華;地塞美松中間體的C_(1,4)脫氫和11α-羥基化[J];生物工程學(xué)報(bào);2000年06期

7 王鑫昕;王少華;李維;李寅;張延平;;細(xì)菌的有機(jī)溶劑耐受機(jī)制[J];生物工程學(xué)報(bào);2009年05期

8 胡純鏗,白鳳武,安利佳;細(xì)胞膜磷脂棕櫚酸含量對(duì)自絮凝顆粒酵母耐酒精能力的影響及作用機(jī)制[J];中國生物工程雜志;2004年08期

9 吳兆亮,胡濱,鄭輝杰,元英進(jìn);兩液相培養(yǎng)中有機(jī)溶劑對(duì)細(xì)胞毒性的研究進(jìn)展[J];生物技術(shù);2000年02期

10 張瑛,王明耀,姚傳義,張寶文,王致萍;固定化簡單節(jié)桿菌轉(zhuǎn)化醋酸可的松動(dòng)力學(xué)[J];天津大學(xué)學(xué)報(bào);1999年01期

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