Scientia Agricultura Sinica ›› 2014, Vol. 47 ›› Issue (2): 324-333.doi: 10.3864/j.issn.0578-1752.2014.02.012

• SOIL & FERTILIZER·WATER-SAVING IRRIGATION·AGROECOLOGY & ENVIRONMENT • Previous Articles     Next Articles

Screening, Identification of Lignin-Degradating Bacillus MN-8 and Its Characteristics in Degradation of Maize Straw Lignin

 LI  Hong-Ya, LI  Shu-Na, WANG  Shu-Xiang, WANG  Quan, ZHU  Bao-Cheng   

  1. College of Life Sciences, Agricultural University of Hebei, Baoding 071000, Hebei
  • Received:2013-03-01 Online:2014-01-15 Published:2013-03-01

RichHTML

2

PDF

803

Cited

14

Abstract: 【Objective】The objective of this study is to obtain the Bacillus with the high lignin-degradation ability and investigate its characteristics in degradation of maize straw lignin, and to provide a basis for microbiological degradation of lignin. 【Method】Bacillus with high ability for degrading lignin was isolated and screened from cattle manure by the decolorization of aniline blue in plate cultivation and the lignin degradation enzyme activity was determined. Among of the obtained Bacillus, the strain with higher ability for degrading lignin was identified through morphology, physiological and biochemical tests, 16S rDNA and gyrB sequence. Then the straw heap fermentation experiment was performed with the inoculation of the bacterial strain obtained above. The changes of lignin peroxidase (LiP), manganese peroxidase (MnP) and the content of lignin in maize straw in the fermentaiton process were monitored to study the lignin-degradation activity of the strain. Its mechanism for degrading lignin was clarified by the GC/MS analysis results of lignin-degradation products. 【Result】 A strain of spore bacterium MN-8 with high ability of lignin-degradation was isolated and screened. Based on the morphology, physiological and biochemical experiments and 16S rDNA sequence analysis, strain MN-8 was identified as Bacillus. The result of 16S rDNA sequence analysis showed that MN-8 was more than 99% similar with Bacillus licheniformis and B. amyloliquefaciens. But the phylogenetic tree based on gyrB sequences showed that strain MN-8 shared 99% identity with B. amyloliquefaciens. Combined with 16S rDNA and gyrB sequence analysis, the strain MN-8 was confirmed as B. amyloliquefaciens. After 16 d incubation in the process of straw heap fermentation, lignin degradation rate was up to 24%. In the two stages of 6-8 d and 10-12 d in the stacking fermentation, the maximum activities of MnP and LiP were appeared. Lignin was degradated apparently in the same stage. And the analysis results of degradation products by GC/MS showed that strain MN-8 could degradate lignin efficiently into some low-molecular-weight matter, such as aromatic compounds 4-hydroxy-3,5-dimethoxy-acetophenone and short chain fatty acids, etc. 【Conclusion】B. amyloliquefaciens MN-8 with high lignin-degradation ability was obtained. It could degradate straw lignin into small molecular substances such as aromatic compounds by the rupture of β-O-4 bond connected between lignin monomer. And its degradation capacity depends on the LiP and MnP.

Key words: lignin-degradation , bacillus , screening , identification , degradation characteristics

[1]Tien M, Kirk T K. Lignin degrading enzyme from the hymenomycete Phenerochaete chrysosporium burds. Science, 1983, 221: 661-663.

[2]Glenn J K, Morgan M A, Mayfield M B, Kuwahara M, Gold M H. An extracellular H2O2-requiring enzyme preparation involved in lignin biodegradation by the white rot basidiomycete Phanerochaete chrysosporium. Biochemical and Biophysical Research Communications, 1983, 114(3): 1077-1083.

[3]Sánchez C. Lignocellulosic residues: biodegradation and bioconversion by fungi. Biotechnology Advances, 2009, 27(2): 185-194.

[4]ten Have R, Teunissen P J. Oxidative mechanisms involved in lignin degradation by white-rot fungi. Chemical Reviews, 2001, 101(11): 3397-3413.

[5]Vicuna R. Bacterial degradation of lignin. Enzyme and Microbial Technology, 1988, 10(11): 646-655.

[6]Zimmermann W. Degradation of lignin by bacteria. Journal of Biotechnology, 1990, 13(2/3): 119-130.

[7]Bugg T D H, Ahmad M, Hardiman E M, Singh R. The emerging role for bacteria in lignin degradation and bio-product formation. Current Opinion in Biotechnology, 2011, 22: 394-400.

[8]Reid I D. Biodegradation of lignin. Canadian Journal of Botany, 1995, 73(S1): 1011-1018.

[9]Kamoda S, Saburi Y. Structural and enzymatical comparison of lignostilbene-α, β-dioxygenase isozymes, I, II, and III, from Pseudomonas paucimobilis TMY1009. Bioscience, Biotechnology, and Biochemistry, 1993, 57(6): 931-934.

[10]Borneman W S, Hartley R D, Morrison W H, Akin D E, Ljungdahl L G. Feruloyl and p-coumaroyl esterase from anaerobic fungi in relation to plant cell wall degradation. Applied Microbiology and Biotechnology, 1990, 33: 345-351.

[11]Kuhad R C, Singh A, Eriksson K E. Microorganisms and enzymes involved in the degradation of plant fiber cell walls. Advances in Biochemical Engineering/Biotechnology, 1997, 57: 45-125.

[12]燕红, 杨谦. 地衣芽孢杆菌对麦麸降解作用的研究. 林产化学与工业, 2007, 27(4): 97-102.

Yan H, Yang Q. Study on the degradation of wheat bran by Bacullus licheniformis. Chemistry and Industry of Forest Products, 2007, 27(4): 97-102. (in Chinese)

[13]燕红, 杨谦, 潘忠诚. 一株地衣芽孢杆菌对稻草降解作用的研究. 浙江大学学报: 农业与生命科学版, 2007, 33(4): 360-366.

Yan H, Yang Q, Pan Z C. Study on degradation of rice straw by Bacillus licheniformis. Journal of Zhejiang University: Agriculture•& Life Science, 2007, 33(4): 360-366. (in Chinese)

[14]燕红, 杨谦. 蜡样芽孢杆菌对稻草的降解作用. 哈尔滨工业大学学报, 2008, 40(8): 1242-1246.

Yan H, Yang Q. Degradation of straw by Bacillus cereus. Journal of Harbin Institute of Technology, 2008, 40(8): 1242-1246. (in Chinese)

[15]丁志刚, 郭亮, 蒋建军, 袁莉莉. 地衣芽孢杆菌对小麦秸秆和稻草品质的影响. 扬州大学学报: 农业与生命科学版, 2010, 31(2): 82-86.

Ding Z G, Guo L, Jiang J J, Yuan L L. Effect of inoculating Bacillus licheniformis on quality of wheat and rice straws. Journal of Yangzhou University: Agricultural and Life Science Edition, 2010, 31(2): 82-86. (in Chinese)

[16]Raj A, Krishna Reddy M M, Chandra R. Identification of low molecular weight aromatic compounds by gas chromatography-mass spectrometry (GC-MS) from kraft lignin degradation by three Bacillus sp.. International Biodeterioration & Biodegradation, 2007, 59: 292-296.

[17]李成翠, 李术娜，朱宝成. 高活性木质素降解菌株T-8的分离、筛选与鉴定. 河北农业大学学报, 2010, 33(6): 57-62.

Li C C, Li S N, Zhu B C. Isolation, screening and identification of the strain T-8 with high lignin-degrading activity. Journal of Agricultural University of Hebei, 2010, 33(6): 57-62. (in Chinese)

[18]王纳贤, 郭晓军, 李术娜, 李春辉, 朱宝成. 产芽孢木质素降解菌株N13的分离与鉴定. 畜牧与饲料科学, 2009(9): 7-9.

Wang N X, Guo X J, Li S N, Li C H, Zhu B C. Screening and identification on bacteria N13 with spore and capability of degrading cellulose. Animal Husbandry and Feed Science, 2009(9): 7-9. (in Chinese)

[19]东秀珠, 蔡妙英. 常见细菌系统鉴定手册. 北京: 科学出版社, 2001: 349-388.

Dong X Z, Cai M Y. Identification of Common Bacterial Systems Manual. Beijing: Science Press, 2001: 349-388. (in Chinese)

[20]程丽娟, 薛泉宏. 微生物学实验技术. 西安: 世界图书出版公司, 2000: 80-83.

Cheng L J, Xue Q H. Experimental Techniques of Microbiology. Xi’an: World Publishing Corporation, 2000: 80-83. (in Chinese)

[21]Kim S B, Yoon J H, Kim H, Lee S T, Park Y H, Goodfellow M. A phylogenetic analysis of the genus Saccharomonospora conducted with 16S rRNA gene sequences. International Journal of Systematic Bacteriology, 1995, 45(2): 351-356.

[22]Rainey F A, Ward-Rainey N, Kroppenstedt R M, Stackebrandt E. The genus Nocardiopsis represents a phylogenetically coherent taxon and a distinct actinomycete lineage: Proposal of Nocardiopsaceae fam. nov.. International Journal of Systematic Bacteriology, 1996, 46(4): 1088-1092.

[23]Robertson B R, Tezuka N, Watanabe M M. Phylogenetic analyses of Synechococcus strains (cyanobacteria) using sequences of 16S rDNA and part of the phycocyanin operon reveal multiple evolutionary lines and reflect phycobilin content. International Journal of Systematic and Evolutionary Microbiology, 2001, 51(3): 861-871.

[24]Yamamoto S, Harayama S. PCR amplification and direct sequencing of gyrB genes with universal primers and their application to the detection and taxonomic analysis of Pseudomonas putida strains. Applied and Environmental Microbiology, 1995, 61(3): 1104-1109.

[25]Saitou N, Nei M. The neighbour-joining method: a new method for reconstructing phylogenetic trees. Molecular Biology and Evolution, 1987, 4(4): 406-425.

[26]潘晓辉. 微波预处理玉米秸秆的工艺研究[D]. 哈尔滨: 哈尔滨工业大学, 2007.

Pan X H. Study on microwave-pretreatment of corn stover[D]. Harbin: Harbin Institute of Technology, 2007. (in Chinese)

[27]Habu N, Komatsu T, Samejima M, Yoshimoto T. Metabolism of a diarylpropane type lignin model compound by Pseudmonas sp. TMY1009. Mokuzai Gakkaishi, 1989, 35: 348-355.

[28]Hammel K E. Extracellular free radical biochemistry of ligninolytie fungi. New Journal of Chemistry, 1996, 20(2): 195-198.

[29]Perez J, Munoz-Dorado J, de la Rubia T, Martinez J. Biodegradation and biological treatments of cellulose, hemicellulose and lignin: an overview. International Microbiology, 2002, 5(2): 53-63.

[30]马登波, 刘紫鹃, 高培基, 王祖农. 木素生物降解的研究进展. 纤维素科学与技术, 1996, 4(1): 1-12.

Ma D B, Liu Z J, Gao P J, Wang Z N. Advance in the research of lignin biodegradation. Journal of Cellulose Science and Technology, 1996, 4(1): 1-12. (in Chinese)
[1] LIN Ping, WANG KaiLiang, YAO XiaoHua, REN HuaDong. Development of DNA Molecular ID in Camellia oleifera Germplasm Based on Transcriptome-Wide SNPs [J]. Scientia Agricultura Sinica, 2023, 56(2): 217-235.
[2] BIAN NengFei, SUN DongLei, GONG JiaLi, WANG Xing, XING XingHua, JIN XiaHong, WANG XiaoJun. Evaluation of Edible Quality of Roasted Peanuts and Indexes Screening [J]. Scientia Agricultura Sinica, 2022, 55(4): 641-652.
[3] WANG MengRui, LIU ShuMei, HOU LiXia, WANG ShiHui, LÜ HongJun, SU XiaoMei. Development of Artificial Inoculation Methodology for Evaluation of Resistance to Fusarium Crown and Root Rot and Screening of Resistance Sources in Tomato [J]. Scientia Agricultura Sinica, 2022, 55(4): 707-718.
[4] SHEN Qian,ZHANG SiPing,LIU RuiHua,LIU ShaoDong,CHEN Jing,GE ChangWei,MA HuiJuan,ZHAO XinHua,YANG GuoZheng,SONG MeiZhen,PANG ChaoYou. Construction of A Comprehensive Evaluation System and Screening of Cold Tolerance Indicators for Cold Tolerance of Cotton at Seedling Emergence Stage [J]. Scientia Agricultura Sinica, 2022, 55(22): 4342-4355.
[5] HUANG Chong,HOU XiangJun. Crop Classification with Time Series Remote Sensing Based on Bi-LSTM Model [J]. Scientia Agricultura Sinica, 2022, 55(21): 4144-4157.
[6] DUAN CanXing,CAO YanYong,DONG HuaiYu,XIA YuSheng,LI Hong,HU QingYu,YANG ZhiHuan,WANG XiaoMing. Precise Characterization of Maize Germplasm for Resistance to Pythium Stalk Rot and Gibberella Stalk Rot [J]. Scientia Agricultura Sinica, 2022, 55(2): 265-279.
[7] YANG JingYa,HU Qiong,WEI HaoDong,CAI ZhiWen,ZHANG XinYu,SONG Qian,XU BaoDong. Consistency Analysis of Classification Results for Single and Double Cropping Rice in Southern China Based on Sentinel-1/2 Imagery [J]. Scientia Agricultura Sinica, 2022, 55(16): 3093-3109.
[8] WANG LuWei,SHEN ZhiJun,LI HeHuan,PAN Lei,NIU Liang,CUI GuoChao,ZENG WenFang,WANG ZhiQiang,LU ZhenHua. Analysis of Genetic Diversity of 79 Cultivars Based on SSR Fluorescence Markers for Peach [J]. Scientia Agricultura Sinica, 2022, 55(15): 3002-3017.
[9] MENG Yu,WEN PengFei,DING ZhiQiang,TIAN WenZhong,ZHANG XuePin,HE Li,DUAN JianZhao,LIU WanDai,FENG Wei. Identification and Evaluation of Drought Resistance of Wheat Varieties Based on Thermal Infrared Image [J]. Scientia Agricultura Sinica, 2022, 55(13): 2538-2551.
[10] ZHANG WeiDong,ZHENG YuJie,GE Wei,ZHANG YueLang,LI Fang,WANG Xin. Identification of Cashmere Dermal Papilla Cells Based on Single- Cell RNA Sequencing Technology [J]. Scientia Agricultura Sinica, 2022, 55(12): 2436-2446.
[11] XU Xiao,REN GenZeng,ZHAO XinRui,CHANG JinHua,CUI JiangHui. Accurate Identification and Comprehensive Evaluation of Panicle Phenotypic Traits of Landraces and Cultivars of Sorghum bicolor (L.) Moench in China [J]. Scientia Agricultura Sinica, 2022, 55(11): 2092-2108.
[12] ZHONG YanPing,SHI LiSong,ZHOU Rong,GAO Yuan,HE YanQing,FANG Sheng,ZHANG XiuRong,WANG LinHai,WU ZiMing,ZHANG YanXin. Establishment of High Efficient Extraction and Detection Technology of Sesamin and Screening of High Sesamin Germplasm [J]. Scientia Agricultura Sinica, 2022, 55(11): 2109-2120.
[13] FAN WenJing,LIU Ming,ZHAO Peng,ZHANG QiangQiang,WU DeXiang,GUO PengYu,ZHU XiaoYa,JIN Rong,ZHANG AiJun,TANG ZhongHou. Screening of Sweetpotato Varieties Tolerant to Low Nitrogen at Seedling Stage and Evaluation of Different Nitrogen Efficiencies [J]. Scientia Agricultura Sinica, 2022, 55(10): 1891-1902.
[14] FENG Chen,FENG LiangShan,LIU Qi,LI HaoRu,ZHENG JiaMing,YANG Ning,BAI Wei,ZHANG Zhe,SUN ZhanXiang. The Degradation Characteristics of Different Plastic Films and Their Effects on Maize Yield in Semi-Arid Area in Western Liaoning [J]. Scientia Agricultura Sinica, 2021, 54(9): 1869-1880.
[15] LIU Chuang,GAO Zhen,YAO YuXin,DU YuanPeng. Functional Identification of Grape Potassium Ion Transporter VviHKT1;7 Under Salt Stress [J]. Scientia Agricultura Sinica, 2021, 54(9): 1952-1963.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
No Suggested Reading articles found!
京ICP备09089781号-30
Copyright 2018 © Editorial office of Scientia Agricultura Sinica
Tel: 86-010-82106279    E-mail: zgnykx@mail.caas.net.cn
Supported by Beijing Magtech Co.Ltd