厌氧微生物培养装置实验教学改革与实践
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摘要
针对现有实验室厌氧微生物培养使用的装置存在通用性差、价格昂贵、易爆炸及使用寿命短等问题,本研究设计、制备了一种模块化多功能实验室厌氧微生物培养装置,并将该装置用于严格厌氧微生物Geobacter sulfurreducens PCA及Geobacter grbiciae与Methanosarcina barkeri 800 co-culture的培养以验证其有效性。实验结果表明,该装置完全满足严格厌氧微生物的培养。设计及制备的厌氧微生物培养装置具有结构简单、体积可变、成本低及使用寿命长等优点,可用于二氧化碳、氮气及H_2/CO_2等气氛厌氧微生物的培养、分离及筛选。该装置为微生物学、环境微生物技术及发酵工程的实验教学创造了良好的实验平台,并取得了良好的实验教学效果。
The existing laboratory devices for anaerobic microbial cultivation either have poor generality or are very expensive,easily explosive,and have short service time. To solve these problems,a new,simple,modularized,and multifunctional device for anaerobic microbial cultivation was developed in this study. The effectiveness of the device was verified by cultivating strict anaerobic microorganism Geobacter sulfurreducens PCA and the co-culture of Geobacter grbiciae and Methanosarcina barkeri 800. The results show that the device completely meets the requirement of strict anaerobic microbial cultivation. The developed device is inexpensive,and has simple structure,changeable value,and long service time. It can be used for the cultivation,isolation,and screening of anaerobic microbe under carbon dioxide,nitrogen or H_2/CO_2 atmosphere. Good results have been obtained in experimental teaching of microbiology,environmental microbial technology,and fermentation engineering through the application of the device in Inner Mongolia University of Science and Technology. The device creates a good experimental platform for experimental teaching of microbiology,environmental microbial technology,and fermentation engineering in Inner Mongolia University of Science and Technology.
The existing laboratory devices for anaerobic microbial cultivation either have poor generality or are very expensive,easily explosive,and have short service time. To solve these problems,a new,simple,modularized,and multifunctional device for anaerobic microbial cultivation was developed in this study. The effectiveness of the device was verified by cultivating strict anaerobic microorganism Geobacter sulfurreducens PCA and the co-culture of Geobacter grbiciae and Methanosarcina barkeri 800. The results show that the device completely meets the requirement of strict anaerobic microbial cultivation. The developed device is inexpensive,and has simple structure,changeable value,and long service time. It can be used for the cultivation,isolation,and screening of anaerobic microbe under carbon dioxide,nitrogen or H_2/CO_2 atmosphere. Good results have been obtained in experimental teaching of microbiology,environmental microbial technology,and fermentation engineering through the application of the device in Inner Mongolia University of Science and Technology. The device creates a good experimental platform for experimental teaching of microbiology,environmental microbial technology,and fermentation engineering in Inner Mongolia University of Science and Technology.
引文
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[10]黎景景,高鑫,李宽钰,等.敲除牙龈卟啉单胞菌菌毛次要组分Fim CDE的载体构建[J].上海口腔医学,2013,22(6):623-627.
[11]王爱宽,秦勇.褐煤本源菌在煤层生物气生成中的微生物学特征[J].中国矿业大学学报,2011,40(6):888-893.
[12]徐隋意.间断多次低温对糖氧剥夺神经元的保护作用[D].广州:南方医科大学,2013.
[13]丁帮璟,李正魁,朱鸿杰,等.河岸带表层土壤的铁氨氧化(Feammox)脱氮机制的探究[EB/OL].[2017-10-27].http://kns.cnki.net/kcms/detail/11.1895.X.20171027.1350.044.html.DOI:10.13227/j.hjkx.201707074.
[14]张祥胜,范红香,许德军.一种简易厌氧生物反应器[P].中国.专利号:ZL201320576109.0.
[15]Tremblay P L,Aklujkar M,Leang C,et al.A genetic system for Geobacter metallireducens:role of the flagellin and pilin in the reduction of Fe(III)oxide[J].Environmental Microbiology Reports,2012,4(1):82-88.
[16]Rotaru A E,Shrestha P M,Liu F,et al.Direct interspecies electron transfer between geobacter metallireducens and methanosarcina barkeri[J].Appl Environ Microbiol,2014,80(15):4599-4605.
[2]李莹,郑世玲,张洪霞,等.产甲烷分离物中Clostridium spp.与Methanosarcina barkeri潜在的种间直接电子传递[J].微生物学通报,2017,44(3):591-600.
[3]孙美洲,金巍,李袁飞,等.瘤胃降解粗纤维产甲烷的厌氧真菌与甲烷菌共培养物的分离鉴定[J].微生物学报,2014,54(5):563-571.
[4]郭亚丽.厌氧生产1,3-丙二醇菌株的分离鉴定及发酵特性研究[D].上海:华东理工大学,2017.
[5]顾风云.SRB培养基氮源及其处理酸性矿山废水条件研究[D].长春:吉林大学,2017.
[6]桂冠,龚文琪,刘晓烨,等.脱硫肠状菌属还原高浓度硫酸盐的试验研究[J].武汉理工大学学报,2013,35(11):138-144.
[7]李亚峰,张文静,马晨曦,等.厌氧氨氧化脱氮性能及主要影响因素的试验[J].沈阳建筑大学学报(自然科学版),2013,29(2):333-337.
[8]侯轩,徐纪茹,辜依海,等.4313份血样细菌培养结果分析[J].西安交通大学学报(医学版),2017,38(6):921-925.
[9]施曼玲,戴赟,章玲.丁酸梭菌培养条件的研究[J].杭州师范大学学报(自然科学版),2001,18(4):38-40.
[10]黎景景,高鑫,李宽钰,等.敲除牙龈卟啉单胞菌菌毛次要组分Fim CDE的载体构建[J].上海口腔医学,2013,22(6):623-627.
[11]王爱宽,秦勇.褐煤本源菌在煤层生物气生成中的微生物学特征[J].中国矿业大学学报,2011,40(6):888-893.
[12]徐隋意.间断多次低温对糖氧剥夺神经元的保护作用[D].广州:南方医科大学,2013.
[13]丁帮璟,李正魁,朱鸿杰,等.河岸带表层土壤的铁氨氧化(Feammox)脱氮机制的探究[EB/OL].[2017-10-27].http://kns.cnki.net/kcms/detail/11.1895.X.20171027.1350.044.html.DOI:10.13227/j.hjkx.201707074.
[14]张祥胜,范红香,许德军.一种简易厌氧生物反应器[P].中国.专利号:ZL201320576109.0.
[15]Tremblay P L,Aklujkar M,Leang C,et al.A genetic system for Geobacter metallireducens:role of the flagellin and pilin in the reduction of Fe(III)oxide[J].Environmental Microbiology Reports,2012,4(1):82-88.
[16]Rotaru A E,Shrestha P M,Liu F,et al.Direct interspecies electron transfer between geobacter metallireducens and methanosarcina barkeri[J].Appl Environ Microbiol,2014,80(15):4599-4605.