《应用于废水处理的说光合细菌优化培养和生长动力学研究.doc》由会员分享，可在线阅读，更多相关《应用于废水处理的说光合细菌优化培养和生长动力学研究.doc（54页珍藏版）》请在三一文库上搜索。
1、应用于废水处理的光合细菌优化培养和生长动力学研究摘 要:本文对应用于废水处理的光合细菌的培养最适温度、光照、pH值、溶解氧等条件进行了较系统的研究,通过正交实验,得出光合细菌生长的最适条件为光照强度3000Lx、微好氧、30e、pH 710。在此基础上,建立了光合细菌在模拟味精废水条件下,以乙酸钠为底物时的生长动力学模型,其参数为饱和常数Ks为0123 g#L-1,最大比生长速率为0104h-1。试验结果表明,该模型能够较好地描述废水处理光合细菌的生长情况。关键词:光合细菌;正交实验;优化培养;动力学模型Study on Optimum Cultivation and Growth Kinet
2、ics of Photosynthetic Bacterial for Wastewater Treatment / XIONG Wan-Abstract:This paper gives a detail study on the optimum growth conditions of temperature, illumination, pH and dissolved oxygen forPSB1With the orthogonal designed experiment, the optimum culture conditions for PSB growth are obtai
3、ned: illumination intensity 3000Lx, faint aerobic, 30e, pH 7101Under the optimum cultivation and the simulated monosodium-glutmate wastewater conditions, a k-inetic model on PSB growth in the substrate of CH3COONa was put up1The parameters in the model are as follows: the saturation con-stantKs0123
4、g#L-1and the maximum specific growth rateLmax0104 h-1respectively1The experiment result shows thatthismodel candescribe the growth data of PSB very well1Key words:photosynthetic bacteria(PSB); orthogonal designed experiment; optimum cultivation; kinetics model 光合细菌(Photosynthetic Bacteria,简称PSB)是自然界
5、中广泛存在且比较古老的细菌类群,是一大类能进行光合作用的原核生物的总称1。近年来,随着人们对PSB形态、结构、生理生化以及生态等特性研究和认识的不断深入,发现光合细菌,特别是其中的红螺菌科能利用多种硫化物或有机物作为其光合作用的供氢体和碳源,在厌氧光照、好氧光照、甚至好氧黑暗环境中都能很好地增殖,且能耐受很高盐度和浓度的有机物,具有很强的分解、去除有机物的能力,显示其在高浓度、高盐度有机废水处理中的独特优势和广阔应用前景,正成为废水处理技术研究的一个新方向2。同时因其菌体富含蛋白质和胡萝卜素,可作为单细胞蛋白应用于养畜业、渔业以及各种食用色素。光合细菌的上述特点,使其吸引着人们对其进行发掘、研
6、究和商品化生产。研究光合细菌的优化培养条件和生长动力学,对其规模化生产和资源化开发利用以及应用于废水处理的光合生物反应系统的设计具有重要意义。1 材料与方法111 菌种来源菌种为本课题组从海水、污泥、池塘水、厌氧生物反应器中分离获得并培养保存3的光合细菌,颜色深红,有微量红色沉淀,其OD660nm大于118,活菌数在11010个#mL-1以上,菌液pH值为816,经鉴定为荚膜红假单胞菌(Rhodopseudomonas capsulata)。112 培养基优化培养实验采用RCVBN扩大培养基4,其组成为:CH3COONa 310 g,(NH4)2SO4110 g,MgSO4012g,NaCl
7、110 g,KH2PO4013 g,K2HPO4015 g,CaCl20105g,酵母膏011 g,微量元素溶液1mL,蒸馏水1000mL。生长动力学实验中以CH3COONa为生长限制性22中国沼气Chian Biogas2004,22(1)底物,考虑到PSB处理味精废水时应适应的环境,使培养基中SO42-浓度控制2%3%,pH值控制在4156范围,其它组分的含量根据CH3COONa底物的浓度按比例调整。微量元素溶液为改良的Imhoff和Truper生长因子溶液,其组成为:EDTA-2Na 2 g,FeSO4#7H2O 012 g,MnCl2#4H2O011 g,H3BO3011 g,CoCl
8、2#6H2O011 g,Zn-Cl2011 g,Na2MoO4#2H2O 0102 g,NiCl2#6H2O 0102 mg,CuCl2#2H2O 0101 g,蒸馏水1000mL。113 培养方法在8个CSTR光合培养器中进行分批培养,培养器体积为50 L。培养器光照根据强度不同的需要分别由三只25W,40W,60W,100W白炽灯泡组合提供,培养器的底部设有穿孔曝气管,根据DO含量间歇开启,培养器内设有温控器。114 测定方法(1)菌体干重:离心后采用恒重法;(2)菌体浊度:采用VIS)7220型分光光度计测OD值(660nm);(3)pH值:采用瑞士Delta 320型酸度计;(4)溶解
9、氧:采用美国YSI)58溶解氧测定仪;(5)光照:由LGC4)2A光照计测定;(6)底物浓度(以CODCr计):采用重铬酸钾法5。2 结果与讨论211 培养条件的优化根据本课题组以前单因素试验的结果6,综合对PSB生长影响较大的四个因素即光照、溶解氧、pH值、温度进行正交试验7,每个因素选取三个水平,选用L9(34)正交表对各因素的试验条件进行优化,具体设计见表1。各次试验菌液的接种量均为5L,即10%,起始OD(660nm)值为01477。培养66 h后测OD值,重复二次,实验结果见表2。 由表可见,各因素对PSB生长的影响主次关系为:光照、溶解氧、温度、pH值。用直观法即可确定光合细菌的优
10、化培养条件为:3,2,2,2,即强光照、微好氧、30e、pH710。表1 正交试验因素表头设计因素水 平1 2 311光照(Lx) 0 1500 300021溶解氧(mg#L-1) 1 0151 001531温度(e) 25 30 3541pH值9 7 5表2 PSB生长情况的正交试验结果及分析试验号因 素1 2 3 4 OD1OD2OD1+OD221 1 1 1 1 01476 01478 014772 1 2 2 2 01808 01825 018163 1 3 3 3 01619 01624 016224 2 1 2 3 01902 01924 019135 2 2 3 1 11161
11、11183 111726 2 3 1 2 11134 11122 111287 3 1 3 2 11590 11605 115978 3 2 1 3 21025 21062 210449 3 3 2 1 21140 21163 21152K1 11915 21987 31649 31801K2 31213 41032 31881 31541K3 51793 31902 31391 31579R 31878 11045 01490 01260212 PSB生长动力学模型的建立21211 PSB的分批培养结果根据上述优化培养的试验结果,采用最适培养条件,即光照强度为3000 Lx、溶解氧为015
12、mg#L-1左右、30e、pH710条件下研究光合细菌生长动力学。在四个(1#4#)相同的光合细菌培养器中,接种量相同,均为10%,在初始底物浓度不同的情况下,研究光合细菌的生长速率和底物的消耗速率。另外,根据本研究大量实验结果,光合细菌的延滞期为6小时,66小时左右后进入衰亡期,所以本实验采用开始6小时后每隔12小时分别测各培养器中底物的浓度(以COD计)以及光合细菌菌体干重值,实验结果见表3。21212 PSB生长动力学模型中参数的估计假设PSB的生长速率只受单一底物CH3COONa浓度的影响,同时不考虑生成产物对光合细菌的抑制作用,可用Monod方程8作为PSB指数生长期和平衡期的生长动
13、力学模型,即PSB的比生长速率L与限制性底物COD的浓度间的关系可用下式表示:L=LmaxSKS+S(1)dXdt=LX(2)-dsdt=1Yx/sdxdt(3)式中:L为光合细菌的比增长速率(h-1);Lmax为最大比增长速率(h-1);S为限制性底物COD的浓度(g#L-1);Ks为光合细菌以乙酸钠为底物的饱和常数(g#L-1);dxdt为光合细菌的生长速率g#(L#h)-1;-dsdt为底物的消耗速率,g#(L#h)-1;Yx/s为光合细菌23中国沼气Chian Biogas2004,22(1)的得率系数(g#g-1)。根据表3中的实验结果,用龙格库塔法求解微分方程组,以实验值与模型估计
14、值总偏差之和最小为目标函数,在C6程序下对培养器1#,2#,3#,4#的实验结果估计的参数值分别为Lmax=01038h-1,01038h-1,0104h-1,01044h-1;Ks=0122g#L-1,0120 g#L-1,0123 g#L-1,0124 g#L-1;Yx/s=01082 g#g-1,01080 g#g-1,01076 g#g-1,01070 g#g-1;目标函数值分别为01000208, 010013, 0100090,0100025。表3 不同时间测得各培养器中的底物浓度和菌体干重值时间t(h)1# 2# 3# 4#COD/mg#L-1菌体干重/mg#L-1COD/mg#
15、L-1菌体干重/mg#L-1COD/mg#L-1菌体干重/mg#L-1COD/mg#L-1菌体干重/mg#L-10 520 117 1436 117 3075 118 6020 1186 508 117 1420 117 3065 119 6000 21018 490 312 1394 318 3029 416 5950 51530 456 610 1336 814 2940 1110 5810 151042 395 1110 1210 1810 2730 2715 5430 421054 296 1910 943 3910 2220 6610 4388 1151066 160 3010 43
16、3 80 1065 15410 1664 3051078 145 3115 406 8117 872 16610 1320 32210213 模拟值与试验值的比较以起始底物浓度COD=4022 mg#L-1在上述条件下培养,在不同时间测得剩余底物浓度和菌体干重,并用上述模型对实验结果进行预测,结果见表4。对模拟值与试验值进行比较,相对误差小于5%,结果见图1。可见,模型能较好地描述强光照微氧条件下以乙酸钠为底物时光合细菌的生长规律。表4 模拟值与试验值比较结果时间t(h)预测值实验值COD/mg#L-1菌体干重/mg#L-1COD/mg#L-1菌体干重/mg#L-10 4022100 1190
17、 4022 1196 4015100 2101 4015 21018 3976108 4196 3982 51130 3879175 12128 3864 131442 3641197 30135 3606 331054 3060112 74157 2892 781666 1684180 179109 1488 1931078 1210 206103 结论(1)温度、光照、pH值和溶解氧等因素会影响PSB的生长,其中光照的强弱影响最大,溶解氧影响次之,pH值影响最小。试验得出PSB生长的适宜条件为强光照、微好氧、温度为30e、pH为710。(2)强光照、微好氧条件下PSB生长动力学模型可用Mo
18、nod方程来模拟,实验测得模型中饱和常数Ks=0123 g#L-1,最大比生长速率=0104 h-1。模型能够较好地描述光合细菌的生长情况。参考文献1 史家梁,翁稣颖,徐亚同1光合细菌在废水处理中的应用及菌体的综合利用J1微生物学通报,1981,4(8):186188.2 俞吉安,李 宝,张承康1应用光合细菌处理高浓度有机废水的新技术J1环境科学,1987,8(3):4752.3 邱宏端,郭养浩,吴陵1光合细菌的分离与生长特性初探J1福州大学学报,1999,27(2)116120.4 朱章玉,俞吉安,林志新1光合细菌的研究及其应用M1上海:上海交通大学出版社,1991.5 国家环保局1水和废水
19、监测分析方法(第三版)M1北京:中国环境科学出版社,1989.6 邱宏端,郭养浩,吴陵1光合细菌的分离与生长特性初探J1福州大学学报,1999,4(2):116120.7 栾军1现代试验设计优化方法M1上海:上海交通大学出版社,1995.8 戚以政,汪叔雄1反应动力学与反应器(第二版)M1化学工业出版社,1999.24中国沼气Chian Biogas2004,22(1)（英文版 ） Two regulations promulgated for implementation is in the party in power for a long time and the rule of law
20、 conditions, the implementation of comprehensive strictly strategic plan, implementation in accordance with the rules and discipline to manage the party, strengthen inner-party supervision of major initiatives. The two regulations supporting each other, the adhere to a positive advocate, focusing on
21、 morality is of Party members and Party leading cadres can see, enough to get a high standard; around the party discipline, disciplinary ruler requirements, listed as negative list, focusing on vertical gauge, draw the party organizations and Party members do not touch the bottom line . Here, the ma
22、in from four square face two party rules of interpretation: the first part introduces two party Revised regulations the necessity and the revision process; the second part is the interpretation of the two fundamental principles of the revision of laws and regulations in the party; the third part int
23、roduces two party regulations modified the main changes and needs to grasp several key problems; the fourth part on how to grasp the implementation of the two regulations of the party. and revised the necessity and revised history of the CPC Central Committee the amendment to the Chinese Communist P
24、arty members and leading cadres honest politics several guidelines and column 1 by 2015 to strengthen party laws and regulations focus. Two party regulations revision work lasted a Years, pooling the wisdom of the whole party, ideological consensus, draw historical experience, respect for the wisdom
25、 of our predecessors, which reflects the unity of inheritance and innovation; follow the correct direction, grasp the limited goals, adhere to the partys leadership, to solve the masses of the people reflect a focus on the problem. The new revision of the and , reflects the partys 18 and the eighth
26、session of the third, the spirit of the fourth plenary session, reflecting the experience of studying and implementing the General Secretary Xi Jinping series of important speech, reflects the partys eighteen years comprehensive strictly practice. (a) revised two regulations of the party need of in
27、based on revised, the promulgation and implementation of January 2010, to strengthen the construction of the contingent of leading cadres play an important role. But with the party to manage the party strictly administering the deepening, has not been able to fully meet the actual needs. Content is
28、too complicated, eight prohibition, 52 are not allowed to hard to remember, and also difficult to put into practice; the second is concisely positive advocated by the lack of prohibited provisions excessive, no autonomy requirements; the third is banned terms and discipline law, both with the party
29、discipline, disciplinary regulations repeat and Criminal law and other laws and regulations repeat; the fourth is to clean the theme is not prominent, not for the existing problems, and is narrow, only needle of county-level leading cadres above. is in 1997 based on revision, in December 2003 the pr
30、omulgation and implementation, to strengthen the construction of the party play very important role. Along with the development of the situation, which many provisions have been unable to fully meet the comprehensive strictly administering the practice needs. One is Ji law, more than half of the pro
31、visions and criminal law and other countries laws and regulations Repetition; two is the political discipline regulations is not prominent, not specific, for violation of the party constitution, damage the authority of Party Constitution of misconduct lack necessary and serious responsibility to pur
32、sue; third is the main discipline for the leading cadres, does not cover all Party members. Based on the above situation, need to and the two is likely to be more relevant regulations first amendment. By revising, really put the authority of Party discipline, the seriousness in the party tree and ca
33、ll up the majority of Party members and cadres of the party constitution of party compasses party consciousness. (II) two party regulations revision process the Central Committee of the Communist Party of China attaches great importance to two regulations revision . Xi Jinping, general books recorde
34、d in the Fifth Plenary Session of the eighth session of the Central Commission for Discipline Inspection, on the revised regulations made clear instructions. According to the central deployment, the Central Commission for Discipline Inspection from 2014 under six months begin study two regulations r
35、evision. The Standing Committee of the Central Commission for Discipline Inspection 4 review revised. Comrade Wang Qishan 14 times held a special meeting to study two regulations revision, amendment clarifies the direction, major issues of principle, path and target, respectively held a forum will l
36、isten to part of the province (area) secretary of the Party committee, Secretary of the Discipline Inspection Commission, part of the central ministries and state organs Department The first party committee is mainly responsible for people, views of experts and scholars and grassroots party organiza
37、tions and Party members. Approved by the Central Committee of the Communist Party of China, on 7 September 2015, the general office of the Central Committee of the Party issued a notice to solicit the provinces (autonomous regions, municipalities) Party, the central ministries and commissions, state
38、 ministries and commissions of the Party (party), the General Political Department of the military, every 3 people organization of Party of two regulations revision opinion. Central Commission for Discipline Inspection of extensive solicitation of opinions, careful study, attracting, formed a revise
39、d sent reviewers. In October 8 and October 12, Central Committee Political Bureau Standing Committee and the Political Bureau of the Central Committee After consideration of the two regulations revised draft. On October 18, the Central Committee of the Communist Party of China formally issued two re
40、gulations. Can say, two laws amendment concentrated the wisdom of the whole party, embodies the party. Second, and revision of the basic principles of two party regulations revision work and implement the partys eighteen, ten eight plenary, the spirit of the Fourth Plenary Session of the Eleventh Ce
41、ntral Committee and General Secretary Xi Jinping important instructions on the revised and , highlighting the ruling party characteristics, serious discipline, the discipline quite in front of the law, based on the current, a long-term, advance as a whole, with Bu Xiuding independent and . Main prin
42、ciple is: first, adhere to the party constitution to follow. The constitution about discipline and self-discipline required specific, awaken the party constitution of party compasses party consciousness, maintaining the authority of the constitution. General Secretary Xi Jinping pointed out that no
43、rules, no side round. Party constitution is the fundamental law, the party must follow the general rules. In early 2015 held the eighth session of the Central Commission for Discipline Inspection Fifth Plenary Session of the 16th Central Committee, Xi Jinping again pointed out that constitution is t
44、he party must follow the general rules, but also the general rules. the revision of the and is Method in adhere to the regulations established for the purpose of combining rule of virtue is to adhere to the party constitution as a fundamental to follow, the constitution authority set up, wake up the
45、 party constitution and party rules the sense of discipline, the party constitution about discipline and self-discipline specific requirements. 4 second is to adhere to in accordance with the regulations governing the party and the party. The Party of rule of virtue de, mainly refers to the partys i
46、deals and beliefs, excellent traditional style. The revised the closely linked to the self-discipline, insisting on the positive initiative, for all members, highlight the vital few, emphasized self-discipline, focusing on the morality, and the majority of Party members and the ideological and moral standards. The revised Ji method separately, Ji, Ji Yan to Method, as a negative list, emphasizing the heteronomy, focusing on vertical gauge.