一株高效凤眼莲脱胶菌的筛选及鉴定

doi:10.11934/j.issn.1673-4831.2017.03.012

生态与农村环境学报 ›› 2017, Vol. 33 ›› Issue (3): 275-280.doi: 10.11934/j.issn.1673-4831.2017.03.012

一株高效凤眼莲脱胶菌的筛选及鉴定

杨庆^1,2, 张超奇¹, 马丽晓¹, 周文兵^1,2, 朱端卫^1,2, 周易勇³, 肖乃东^1,2

1. 华中农业大学微量元素研究中心生态与环境工程研究室, 湖北武汉 430070;
2. 生猪健康养殖协同创新中心, 湖北武汉 430070;
3. 淡水生态与生物技术国家重点实验室, 湖北武汉 430072

收稿日期:2016-06-17 出版日期:2017-03-25 发布日期:2017-03-25
通讯作者: 周文兵,通信作者,E-mail:zhouwb@mail.hzau.edu.cn E-mail:zhouwb@mail.hzau.edu.cn
作者简介:杨庆(1991-),女,湖北孝感人,硕士生,研究方向为农业固体废弃物资源化利用。E-mail:jyq805@126.com
基金资助:
中央高校基本科研业务费专项（2014PY061，2015BQ013，2016PY049）；武汉市关键技术攻关计划（2015020202010126）；教育部留学回国人员科研启动基金；国家水体污染控制与治理科技重大专项（2012ZX07104-001）；淡水生态与生物技术国家重点实验室开放课题（2016FB19）

Screening and Identification of an Efficient Strain of Bacteria Degumming Eichhornia crassipes

YANG Qing^1,2, ZHANG Chao-qi¹, MA Li-xiao¹, ZHOU Wen-bing^1,2, ZHU Duan-wei^1,2, ZHOU Yi-yong³, XIAO Nai-dong^1,2

1. Laboratory of Eco-Environmental Engineering Research, Microelement Research Center, Huazhong Agricultural University, Wuhan 430070, China;
2. The Cooperative Innovation Center for Sustainable Pig Production of Hubei Province, Wuhan 430070, China;
3. State Key Laboratory of Freshwater Ecology and Biotechnology, Wuhan 430072, China

Received:2016-06-17 Online:2017-03-25 Published:2017-03-25

摘要/Abstract

摘要：

预处理是生物质能源产品制备中最昂贵、最关键的环节，生物预处理因环境友好和能耗低而受到广泛关注。为获得较好的生物预处理菌株，以脱胶效率为出发点筛选菌株，从凤眼莲（Eichhornia crassipes）生长环境介质中分离菌株，通过富集培养、刚果红平板染色法初筛，获得一株对凤眼莲脱胶效果较好的A₁菌株。在最佳条件下A₁菌株分泌的果胶酶活性、木聚糖酶活性和纤维素酶活性分别为3 925.00、8 331.67和4 883.62 nmol·s^-1·mL^-1；A₁菌株对凤眼莲的1次脱胶减重率为33.37%，3次连续脱胶的累计减重率为49.26%。通过16S rRNA序列鉴定，构建系统发育树分析，该菌株为解淀粉芽孢杆菌（Bacillus amyloliquefaciens）。

关键词: 脱胶, 凤眼莲, 筛选, 解淀粉芽孢杆菌

Abstract:

Pretreatment is the most expensive and critical step of converting biomass into energy, while biological pretreatment has attracted extensive attention for its environmental friendliness and low energy consumption. In order to find a bacteria efficient for use in biological pretreatment, degumming efficiency was chosen as the scale for strain screening. A strain, coded as A₁, was isolated from the environmental media where Eichhornia crassipes grew through enrichment culture and primary screening with the Congo red plate staining method, and tested to be a strain quite efficient in degumming water hyacinth. Under the optimal conditions, Strain A₁ could secrete pectinase, xylanase and cellulase as high as 3 925.00, 8 331.67 and 4 883.62 nmol·s^-1·mL^-1, respectively, in activity. When used in pretreatment, it could reduce the weight of water hyacinth by 33.37% through one round of degumming, and cumulatively by 49.26% after three rounds of degumming. The strain was identified as Bacillus amyloliquefaciens by means of 16S rRNA sequencing and phylogenetic tree analysis.

Key words: degumming, Eichhornia crassipes, screening, Bacillus amyloliquefaciens

中图分类号:

X172

杨庆, 张超奇, 马丽晓, 周文兵, 朱端卫, 周易勇, 肖乃东. 一株高效凤眼莲脱胶菌的筛选及鉴定[J]. 生态与农村环境学报, 2017, 33(3): 275-280.

YANG Qing, ZHANG Chao-qi, MA Li-xiao, ZHOU Wen-bing, ZHU Duan-wei, ZHOU Yi-yong, XIAO Nai-dong. Screening and Identification of an Efficient Strain of Bacteria Degumming Eichhornia crassipes[J]. Journal of Ecology and Rural Environment, 2017, 33(3): 275-280.

参考文献

[1] NIGAM J N.Bioconversion of Water Hyacinth (Eichhornia crassipes) Hemicellulose Acid Hydrolysate to Motor Fuel Ethanol by Xylose-Fermenting Yeast[J].Journal of Biotechnology,2002,97(2):107-116.
[2] MALIK A.Environmental Challenge Vis a Vis Opportunity:The Case of Water Hyacinth[J].Environment International,2007,33(1):122-138.
[3] BHATTACHARYA A,KUMAR P.Water Hyacinth as a Potential Biofuel Crop[J].Electronic Journal of Environmental Agricultural and Food Chemistry,2010,9(1):112-122.
[4] DING S Y,HIMMEL M E.The Maize Primary Cell Wall Microfibril:A New Model Derived From Direct Visualization[J].Journal of Agricultural and Food Chemistry,2006,54(3):597-606.
[5] 赵晨,崔馨月,刘广青,等.提高玉米秸秆厌氧消化产气能力的预处理技术研究进展[J].生态与农村环境学报,2015,31(5):640-646.[ZHAO Chen,CUI Xin-yue,LIU Guang-qing,et al.Recent Progress in Study on Technology for Pretreating Corn Stalk to Improve Biogas Production Capacity of Anaerobic Digestors[J].Journal of Ecology and Rural Environment,2015,31(5):640-646.]
[6] WYMAN C E.What Is (and Is Not) Vital to Advancing Cellulosic Ethanol[J].Trends Biotechnology,2007,25(4):153-157.
[7] 李萍,韦秀丽,蒋滔,等.不同预处理方式对玉米秸秆结构及产气特性的模拟研究[J].生态与农村环境学报,2014,30(1):84-89.[LI Ping,WEI Xiu-li,JIANG Tao,et al.Simulation of Structure and Biogas Production Properties of Corn Stalk Relative to Pretreatment Method[J].Journal of Ecology and Rural Environment,2014,30(1):84-89.]
[8] HIMMEL M E,DING S Y,JOHNSON D K,et al.Biomass Recalcitrance:Engineering Plants and Enzymes for Biofuels Production[J].Science,2007,315(5813):804-807.
[9] MOSIER N,WYMAN C,DALE B,et al.Features of Promising Technologies for Pretreatment of Lignocellulosic Biomass[J].Bioresource Technology,2005,96(6):673-686.
[10] YANG B,WYMAN C E.Pretreatment:The Key to Unlocking Low-Cost Cellulosic Ethanol[J].Biofuels Bioproducts and Biorefining,2008,2(2):26-40.
[11] KADAM K L,CHIN C Y,BROWN L W.Flexible Biorefinery for Producing Fermentation Sugars,Lignin and Pulp From Corn Stover[J].Journal of Industrial Microbiology and Biotechnology,2008,35(5):331-341.
[12] 郑来久,刘剑宇.黄红麻微生物脱胶影响因素及机理研究[J].东华大学学报(自然科学版),2004,30(3):66-70.[ZHENG Lai-jiu,LIU Jian-yu.Influencing Factors and Mechanism of Microbial Degumming of Jute/Kenaf[J].Journal of Donghua University(Natural Science),2004,30(3):66-70.]
[13] 杨丹,王燕,王茜,等.蜡样芽孢杆菌用于苎麻脱胶研究[J].中国农学通报,2015,31(3):260-264.[YANG Dan,WANG Yan,WANG Qian,et al.Research on Bacillus cereus for the Degumming of Ramie[J].Chinese Agricultural Science Bulletin,2015,31(3):260-264.]
[14] 汪学军,闵长莉,杨艳,等.大麻微生物脱胶特异放线菌菌株的筛选与鉴定[J].纺织学报,2014,35(11):102-106.[WANG Xue-jun,MIN Chang-li,YANG Yan,et al.Isolation and Identification of Actinomycete Strain DM182 for Microbial Degumming of Hemp[J].Journal of Textile Research,2014,35(11):102-106.]
[15] 董政娥,丁若垚,郑磊,等.亚麻粗纱碱性脱胶菌株的筛选鉴定及脱胶效果[J].印染助剂,2013,30(10):16-19.[DONG Zheng-e,DING Ruo-yao,ZHENG Lei,et al.Screening and Identification of Alkalophilic Degumming Strains of Flax Roving and Evaluation of Its Effects[J].Textile Auxiliaries,2013,30(10):16-19.]
[16] 蓝广芊,左伟东,许平震,等.苎麻生物脱胶特异菌株的筛选与鉴定[J].纺织学报,2010,31(8):56-60.[LAN Guang-qian,ZUO Wei-dong,XU Ping-zhen,et al.Screening and Identification of a Bacterium Strain for Specific Ramie Degumming[J].Journal of Textile Research,2010,31(8):56-60.]
[17] 陈青,韩双艳,郑穗平,等.剑麻生物制浆脱胶菌株的筛选与鉴定及初步应用[J].农业工程学报,2008,24(6):277-281.[CHEN Qing,HAN Shuang-yan,ZHENG Sui-ping,et al.Screening and Identifying Strain of Bio-Pulping and Degumming of the Sisal and Its Preliminary Application[J].Transactions of the CSAE,2008,24(6):277-281.]
[18] 马丽晓.凤眼莲秸秆生物脱胶的研究[D].武汉:华中农业大学,2009.[MA Li-xiao.Study on the Biological Degumming to Straw of Eichhornia crassipes[D].Wuhan:Huazhong Agricultural University,2009.]
[19] 董鹤娟,丁轲,恒子钤,等.一株土壤源高产纤维素酶芽孢杆菌的分离与鉴定[J].家畜生态学报,2013,34(6):48-52.[DONG He-juan,DING Ke,HENG Zi-qin,et al.Isolation and Identifitication of a High Yield Cellulase bacillus Strain From Soil[J].Acta Ecologiae Animalis Domastici,2013,34(6):48-52.]
[20] 金迪,彭清净,易浪波,等.一株纤维素降解细菌的筛选、鉴定及产酶条件分析[J].中国微生态学杂志,2010,22(4):289-295.[JIN Di,PENG Qing-jing,YI Lang-bo,et al.Isolation and Identification of a Cellulose-Decomposing Bacterium and Its Cellulase-Producing Conditions[J].Chinese Journal of Microecology,2010,22(4):289-295.]
[21] 金钢.大麻脱胶方法的研究进展[J].南京林业大学学报(自然科学版),2009,33(4):140-144.[JIN Gang.Research Progress of Degumming Technology of Cannabis Sativa[J].Journal of Nanjing Forestry University (Natural Sciences Edition),2009,33(4):140-144.]
[22] 赵斌,何绍江.微生物学实验[M].北京:科学出版社,2002:38-39.[ZHAO Bin,HE Shao-jiang.Laboratory Experiments in Microbiology[M].Beijing:Science Press,2002:38-39.]
[23] 关晓欢,姜华.解淀粉芽孢杆菌研究进展[J].生物技术世界,2013(1):4-9.[GUAN Xiao-huan,JIANG Hua.The Research on Development of Bacillus amyloliquefaciens[J].Biotech World,2013(1):4-9.]
[24] 翟秋梅,薛卫巍,薛永常,等.枯草芽孢杆菌FM208849产果胶酶发酵条件的优化[J].大连工业大学学报,2010,29(2):93-97.[ZHAI Qiu-mei,XUE Wei-wei,XUE Yong-chang,et al.The Optimization of Pectinase Fermentation From Bacillus subtilis FM208849[J].Journal of Dalian Polytechnic University,2010,29(2):93-97.]
[25] 于丽萍,郭立姝,殷博,等.枯草芽孢杆菌(Bacillus subtilis) HW201的酶学特性及沤麻实验研究[J].黑龙江科学,2013,4(2):18-22.[YU Li-ping,GUO Li-shu,YIN Bo,et al.The Identification and Enzymatic Properties of the Bacillus subtilis Strain for Flax Retting[J].Heilongjiang Science,2013,4(2):18-22.]
[26] 吴皓琼,曹亚彬,郭立姝,等.枯草芽孢杆菌HW201的亚麻生物脱胶条件优化[J].中国麻业科学,2010,32(5):265-269.[WU Hao-qiong,CAO Ya-bin,GUO Li-shu,et al.The Condition Optimization of Retting Flax With Bacillus subtilis HW201[J].Plant Fiber Sciences in China,2010,32(5):265-269.]
[27] 李超,吴为中,吴伟龙,等.解淀粉芽孢杆菌对鱼腥藻的抑藻效果分析与机理初探[J].环境科学学报,2011,31(8):1602-1608.[LI Chao,WU Wei-zhong,WU Wei-long,et al.Ability of Bacillus amyloliquefaciens to Inhibit Growth of Anabaena flosaquae and the Inhibition Mechanism[J].Acta Scientiae Circumstantiae,2011,31(8):1602-1608.]

一株高效凤眼莲脱胶菌的筛选及鉴定

Screening and Identification of an Efficient Strain of Bacteria Degumming Eichhornia crassipes

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