微生物燃料电池专利

一种芽孢杆菌的产电特性及其在微生物燃料电池中的应用一、技术领域本发明涉及一种革兰氏阳性蜡样芽孢杆菌（Bacillus cereus）的产电特性及其在微生物燃料电池中的应用研究。

二、背景技术随着人类的进步与发展，对能源的需求愈加强烈。

为了解决能源短缺问题，人类在积极寻求新型能源方式。

在能源的寻求过程中，科学家把眼光投向了微生物，利用微生物产生电能，即微生物燃料电池（Microbial Fuel Cell，MFC）。

早在1911 年英国植物学家Potter 发现微生物可以产生电流，生物燃料电池研究由此开始，但在以后的发展中，有关MFC 的研究一直进展缓慢。

早期的MFC是用发酵型的酵母和细菌进行制电。

近几年来，随着产电微生物的(Electricigens)发现，科学家发明了无需使用电子传递中间体的微生物电池。

在MFC这个系统中，产电微生物是核心要素。

产电微生物就是那些能够在厌氧条件下完全氧化有机物成CO2，然后把氧化过程中产生的电子通过电子传递链传递到电极上产生电流的微生物，同时微生物在电子传递过程中获得能量支持生长。

在废水中，一般含有丰富的有机物质。

以这种新型微生物呼吸方式为基础的MFC可以同时进行废水处理和生物发电，有望可以把废水处理发展成一个有利可图的产业，是MFC最有发展前景的方向。

随着MFC 在未来的不断进步和发展，将会有越来越多的应用领域。

微生物产电的能力差异很大，产电微生物决定着MFC的功能与应用，是MFC 系统的核心。

高产电微生物的不断被发现，使得MFC 的内涵发生了根本性的变革，同时也展现了MFC的广阔应用前景。

已经报道的产电微生物多数为铁还原菌，即以Fe(Ⅲ)为呼吸链的最终电子受体的微生物。

三、发明内容本发明的目的是提供从自然界分离出具有高活性产电能力的短小芽孢杆菌（Bacillus pumilus）及其在微生物燃料电池方面的应用研究，该菌具有产电能力强，能够利用有机废水进行产电。

在微生物燃料电池中具有很高的产电活性，具有广阔的应用前景和理论研究价值。

本发明的技术方案如下：一种革兰氏阳性菌短小芽孢杆菌（Bacillus pumilus），菌体为杆状，直径为0.6~0.7μm，菌长为2.0~3.0μm。

本发明的短小芽孢杆菌（Bacillus pumilus）是分离自南京大学生命科学学院门前的有机废水中，在微生物燃料电池中以葡萄糖为电子供体进行培养，其生物学特征如下：形态学特性：菌体细杆状，菌体直，两端圆形，一般为0.6~0.7μm×2.0~3.0μm，革兰氏阳性。

菌落表面光滑，有光泽，不透明，乳白色，边缘整齐。

培养特性生理生化特征特征结果碳源利用特征结果明胶液化–葡萄糖+淀粉水解–乳糖–糊精–麦芽糖–尿素水解+ 纤维二糖–分解七叶苷+ 血清菊糖–分解胆汁七叶苷+ 蔗糖+硝酸盐（产气）+ L-阿拉伯糖–西蒙氏枸橼酸盐–阿拉伯醇–利用醋酸盐–D-核糖–利用DNA –木糖–苯丙氨酸脱羧酶–木糖醇–鸟氨酸脱羧酶–半乳糖+赖氨酸脱羧酶–果糖+精氨酸脱羧酶+ 山梨醇–精氨酸双水解酶+ 山梨糖–亚硝酸盐（产气）–卫矛醇–氰化钾生长+ 蜜二糖–硫化氢产生+ 鼠李糖–乙酰胺–蕈糖–半固体琼脂–肌醇–N-乙酰葡萄糖胺–丙二酸盐–丙酮酸钠–酒石酸盐–氧化发酵试验+ 葡萄糖（产气）–16S rRNA基因序列本发明菌株的16S rRNA基因序列如下AAGGGAGCTTGCTCCCGGA TGTTAGCGGCGGACGGGTGAGTAACACGTGGGTAACCT GCCTGTAAGACTGGGA TAACTCCGGGAAACCGGAGCTAATACCGGATAGTTCCTTGAA CCGCA TGGTTCAAGGATGAAAGACGGTTTCGGCTGTCACTTACAGA TGGACCCGCGGC GCATTAGCTAGTTGGTGAGGTAACGGCTCACCAAGGCGACGATGCGTAGCCGACCTGA GAGGGTGATCGGCCACACTGGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGC AGTAGGGAATCTTCCGCAA TGGACGAAAGTCTGACGGAGCAACGCCGCGTGAGTGAT GAAGGTTTTCGGA TCGTAAAGCTCTGTTGTTAGGGAAGAACAAGTGCAAGAGTAACTG CTTGCACCTTGACGGTACCTAACCAGAAAGCCACGGCTAACTACGTGCCAGCAGCCGC GGTAA TACGTAGGTGGCAAGCGTTGTCCGGAA TTATTGGGCGTAAAGGGCTCGCAGGC GGTTTCTTAAGTCTGA TGTGAAAGCCCCCGGCTCAACCGGGGAGGGTCA TTGGAAACT GGGAAACTTGAGTGCAGAAGAGGAGAGTGGAA TTCCACGTGTAGCGGTGAAA TGCGT AGAGATGTGGAGGAACACCAGTGGCGAAGGCGACTCTCTGGTCTGTAACTGACGCTG AGGAGCGAAAGCGTGGGGAGCGAACAGGATTAGATACCCTGGTAGTCCACGCCGTAA ACGATGAGTGCTAAGTGTTAGGGGGTTTCCGCCCCTTAGTGCTGCAGCTAACGCATTAA GCACTCCGCCTGGGGAGTACGGTCGCAAGACTGAAACTCAAAGGAATTGACGGGGGC CCGCACAAGCGGTGGAGCATGTGGTTTAA TTCGAAGCAACGCGAAGAACCTTACCAG GTCTTGACATCCTCTGACAACCCTAGAGA TAGGGCTTTCCCTTCGGGGACAGAGTGAC AGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGA TGTTGGGTTAAGTCCCGCAAC GAGCGCAACCCTTGA TCTTAGTTGCCAGCATTCAGTTGGGCACTCTAAGGTGACTGCC GGTGACAAACCGGAGGAAGGTGGGGA TGACGTCAAATCATCATGCCCCTTATGACCTG GGCTACACACGTGCTACAA TGGACAGAACAAAGGGCTGCGAGACCGCAAGGTTTAGC CAATCCCACAAATCTGTTCTCAGTTCGGATCGCAGTCTGCAACTCGACTGCGTGAAGC TGGAATCGCTAGTAA TCGCGGATCAGCATGCCGCGGTGAATACGTTCCCGGGCCTTGTA CACACCGCCCGTCACACCACGAGAGTTTGCAACACCCGAAGTCGGTGAGGTAACCTTT ATGGAGCCAGCCGCTAGAGAGTTGGCTG通过检索GenBank,本发明菌株的16S rRNA基因相似性如下表所示FJ592172.1Bacillus aerophilus strain BH-11 16S ribosomal RNA genes,partial sequence 258699%100%JF309254.1Bacillus sp. 34386DBRRJ 16S ribosomal RNA gene, partialsequence 2584 99%100%JF309253.1Bacillus sp. 34386CBRRJ 16S ribosomal RNA gene, partial 2584 99% 100sequence %JF309252.1Bacillus sp. 34386BBRRJ 16S ribosomal RNA gene, partialsequence 2584 99%100%JF309251.1Bacillus sp. 34386ABRRJ 16S ribosomal RNA gene, partialsequence 2584 99%100%JF309248.1Bacillus sp. 3429DBRRJ 16S ribosomal RNA gene, partialsequence 2584 99%100%JF309246.1Bacillus sp. 3429BBRRJ 16S ribosomal RNA gene, partialsequence 2584 99%100%JF309245.1Bacillus sp. 3429ABRRJ 16S ribosomal RNA gene, partialsequence 2584 99%100%JF262083.1Bacillus sp. GC-4 16S ribosomal RNA gene, partial sequence 2584 99% 100 %HQ916742.1Bacillus sp. Bg-2 16S ribosomal RNA gene, partial sequence 2584 99% 100 %HQ891970.1Bacillus subtilis strain MM113 16S ribosomal RNA gene, partial sequence 2584 99%100%JF313264.1Bacillus sp. NIB6 16S ribosomal RNA gene, partial sequence 2584 99% 100 %JF313263.1Bacillus sp. NIB4 16S ribosomal RNA gene, partial sequence 2584 99% 100 %HQ202562.1Bacillus sp. TZQ28 16S ribosomal RNA gene, partial sequence 2584 99% 100 %HQ202550.1Bacillus sp. RHH58 16S ribosomal RNA gene, partial sequence 2584 99% 100 %HQ143571.1Bacillus subtilis strain YT2 16S ribosomal RNA gene, partialsequence 2584 99%100%HQ143665.1Bacillus altitudinis strain ZAH3 16S ribosomal RNA gene,partial sequence 2584 99%100%HQ143613.1Bacillus sp. strain RHH15 16S ribosomal RNA gene, partialsequence 2584 99%100%HQ143591.1Bacillus pumilus strain FGH7 16S ribosomal RNA gene, partial sequence 2584 99%100%HQ427437.1Uncultured bacterium clone RU06182008D2 16S ribosomalRNA gene, partial sequence 2584 99%100%HQ704708.1Bacillus sp. C250 16S ribosomal RNA gene, partial sequence 2584 99% 99% HQ704706.1Bacillus sp. S244 16S ribosomal RNA gene, partial sequence 2584 99% 99%HQ873709.1Bacterium fjat-scb-3 16S ribosomal RNA gene, partial sequence 2584 99% 100 %HM771663.1Bacillus sp. PPB8 MB8 16S ribosomal RNA gene, partialsequence 2584 99%100%HQ317196.1Bacillus pumilus strain DYJL55 16S ribosomal RNA gene,partial sequence 2584 99%100%HQ317195.1Bacillus pumilus strain DYJL54 16S ribosomal RNA gene,partial sequence 2584 99%100%HQ317176.1Bacillus pumilus strain DYJL-C 16S ribosomal RNA gene,partial sequence 2584 99%100%HQ246228.1Bacillus sp. 8A18S6 16S ribosomal RNA gene, partial sequence 2584 99% 100 %HQ236061.1Bacillus altitudinis strain TAZ1-5 16S ribosomal RNA gene,partial sequence 2584 99%100%HQ236060.1Bacillus altitudinis strain TAZ1-2 16S ribosomal RNA gene,partial sequence 2584 99%100%HQ236053.1Bacillus altitudinis strain SBG1-7 16S ribosomal RNA gene,partial sequence 2584 99%100%HQ236039.1Bacillus altitudinis strain SBD1-9 16S ribosomal RNA gene,partial sequence 2584 99%100%HQ222359.1Bacillus sp. H20(2010b) 16S ribosomal RNA gene, partialsequence 2584 99%100%HQ222353.1Bacillus sp. H14(2010) 16S ribosomal RNA gene, partialsequence 2584 99%100%HQ222352.1Bacillus sp. H13(2010) 16S ribosomal RNA gene, partialsequence 2584 99%100%HQ222351.1Bacillus sp. H12(2010) 16S ribosomal RNA gene, partialsequence 2584 99%100%HQ222350.1Bacillus sp. H11(2010) 16S ribosomal RNA gene, partialsequence 2584 99%100%HQ222345.1Bacillus sp. H6(2010) 16S ribosomal RNA gene, partialsequence 2584 99%100%HQ222341.1Bacillus sp. H2(2010) 16S ribosomal RNA gene, partialsequence 2584 99%100%HQ222340.1Bacillus sp. H1(2010b) 16S ribosomal RNA gene, partialsequence 2584 99%100%HQ222334.1Bacillus sp. L12(2010) 16S ribosomal RNA gene, partialsequence 2584 99%100%HQ222333.1Bacillus sp. L11(2010) 16S ribosomal RNA gene, partialsequence 2584 99%100%HQ222331.1Bacillus sp. L9(2010) 16S ribosomal RNA gene, partial sequence 2584 99% 100 %HQ222330.1Bacillus sp. L8(2010) 16S ribosomal RNA gene, partial sequence 2584 99% 100 %HQ222329.1Bacillus sp. L7(2010) 16S ribosomal RNA gene, partial sequence 2584 99% 100 %HQ222327.1Bacillus sp. L5(2010) 16S ribosomal RNA gene, partial sequence 2584 99% 100 %HQ222326.1Bacillus sp. L4(2010) 16S ribosomal RNA gene, partial sequence 2584 99% 100 %HM104375.1Bacillus sp. AMP-9 16S ribosomal RNA gene, partial sequence 2584 99% 100 %HQ164542.1Bacillus pumilus strain sk-1 16S ribosomal RNA gene, partial sequence 2584 99%100%HQ124006.1Bacillus pumilus strain MTCC 7514 16S ribosomal RNA gene, partial sequence 2584 99%100%HM582688.1Bacillus altitudinis strain AP-MSU 16S ribosomal RNA gene, partial sequence 2584 99%100%HM543166.1Bacillus pumilus strain B50 16S ribosomal RNA gene, partial sequence 2584 99%100%HM242289.1Bacillus sp. HaiN-1 16S ribosomal RNA gene, partial sequence 2584 99% 100 %HM246684.1Bacillus sp. Rb-ZD-1 16S ribosomal RNA gene, partial sequence 2584 99% 100 %FN997624.1Bacillus pumilus partial 16S rRNA gene, strain MS28 2584 99% 100 %FN997623.1Vibrio parahaemolyticus partial 16S rRNA gene, strain MS27 2584 99% 100 %HM224386.1Bacillus pumilus strain B1 16S ribosomal RNA gene, partialsequence 2584 99%100%FN666893.1Bacillus sp. 3LF 48P partial 16S rRNA gene, strain 3LF 48P 2584 99% 100 %GU201862.1Bacillus pumilus strain Qtx-10 16S ribosomal RNA gene, partial sequence 2584 99%100%FN668692.1Bacillus altitudinis partial 16S rRNA gene, isolate GVC11 2584 99% 100 %GU332620.1Bacillus sp. BG2-2 16S ribosomal RNA gene, partial sequence 2584 99% 100 %GU332611.1Bacillus sp. AD3-15 16S ribosomal RNA gene, partial sequence 2584 99% 100 %GU332605.1Bacillus sp. BT1-2 16S ribosomal RNA gene, partial sequence 2584 99% 100 %GU332600.1Bacillus pumilus strain BZ3-10 16S ribosomal RNA gene, partial sequence 2584 99%100%GU217692.1Bacillus sp. ljh-2 16S ribosomal RNA gene, partial sequence 2584 99% 100 %partial sequence %GU125636.1Bacillus altitudinis strain IMAU80219 16S ribosomal RNA gene, partial sequence 2584 99%100%GU125627.1Bacillus pumilus strain IMAU80210 16S ribosomal RNA gene, partial sequence 2584 99%100%GU125624.1Bacillus pumilus strain IMAU80207 16S ribosomal RNA gene, partial sequence 2584 99%100%GU084168.1Bacillus pumilus strain SYBC-W 16S ribosomal RNA gene,partial sequence 2584 99%100%GQ903423.1Bacillus pumilus strain XJSL5-4 16S ribosomal RNA gene,partial sequence 2584 99%100%FJ817343.1Bacillus pumilus strain P5-50/08C1 16S ribosomal RNA gene, partial sequence 2584 99%100%GQ465935.1Bacillus subtilis strain N43 16S ribosomal RNA gene, partialsequence 2584 99%100%FJ763649.1Bacillus pumilus strain S68 16S ribosomal RNA gene, partial sequence 2584 99%100%GQ375798.1Bacillus pumilus strain YQQ18 16S ribosomal RNA gene, partial sequence 2584 99%100%GQ375796.1Bacillus pumilus strain YQQ10 16S ribosomal RNA gene, partial sequence 2584 99%100%GQ375793.1Bacillus pumilus strain DZQ18 16S ribosomal RNA gene, partial sequence 2584 99%100%GQ375792.1Bacillus sp. DZQ11 16S ribosomal RNA gene, partial sequence 2584 99% 100 %GQ375785.1Bacillus pumilus strain DZH2 16S ribosomal RNA gene, partial sequence 2584 99%100%GQ375784.1Bacillus pumilus strain DZH1 16S ribosomal RNA gene, partial sequence 2584 99%100%GQ360038.1Bacillus subtilis strain FQ06 16S ribosomal RNA gene, partial sequence 2584 99%100%GQ284505.1Bacillus sp. TSSAS2-44 16S ribosomal RNA gene, partialsequence 2584 99%100%GQ284500.1Bacillus sp. TSSAS2-12 16S ribosomal RNA gene, partialsequence 2584 99%100%GQ284397.1Bacillus sp. TSWCS4 16S ribosomal RNA gene, partial sequence 2584 99% 100 %GQ289128.1Bacterium Kye2 16S ribosomal RNA gene, partial sequence 2584 99% 100 %GQ199752.1Bacillus sp. 210_50 16S ribosomal RNA gene, partial sequence 2584 99% 100 %sequence %FJ842658.1Bacillus sp. DBTMGS2 16S ribosomal RNA gene, partialsequence 2584 99%100%FJ615523.1Bacillus sp. 41KBZ 16S ribosomal RNA gene, partial sequence 2584 99% 100 %FJ615522.1Bacillus sp. 41KAZ 16S ribosomal RNA gene, partial sequence 2584 99% 100 %FJ615521.1Bacillus sp. 28KZ 16S ribosomal RNA gene, partial sequence 2584 99% 100 %FJ611965.1Bacillus altitudinis strain GCH-2 16S ribosomal RNA gene,partial sequence 2584 99%100%FJ594456.1Bacillus sp. BSFA18-5 16S ribosomal RNA gene, partialsequence 2584 99%100%FM955870.1Bacillus altitudinis 16S rRNA gene, strain Asd M5-2B 2584 99% 100 %FJ571618.1Bacillus sp. BSFA18-4 16S ribosomal RNA gene, partialsequence 2584 99%100%FJ544367.1Bacillus pumilus strain st9 16S ribosomal RNA gene, partialsequence 2584 99%100%FJ495143.1Bacillus sp. BSFA18-3 16S ribosomal RNA gene, partialsequence 2584 99%100%FJ495142.1Bacillus sp. BSFA18-2 16S ribosomal RNA gene, partialsequence 2584 99%100%GU726185.1Bacillus sp. KZ_AalM_Mm3 16S ribosomal RNA gene,complete sequence2580 99% 99% AB374299.1Bacillus sp. FP303 gene for 16S rRNA, partial sequence 2580 99% 99% 表显示，本发明的菌株的16S rRNA基因在GenBank中，和Bacillus aerophilus，Bacillus subtilis，Bacillus altitudinis，Bacillus pumilus有着完全相同序列，经生理生化鉴定，为Bacillus pumilus。