Study on the Isolation,Identification of Salt-tolerant Cellulose-Degrading Strains and Optimization of Its Strain Ratio

doi:10.6048/j.issn.1001-4330.2017.12.016

Xinjiang Agricultural Sciences ›› 2017, Vol. 54 ›› Issue (12): 2282-2292.DOI: 10.6048/j.issn.1001-4330.2017.12.016

Previous Articles Next Articles

Study on the Isolation,Identification of Salt-tolerant Cellulose-Degrading Strains and Optimization of Its Strain Ratio

WANG Xu-hui^1,2, YE Kai², XU Xin³, Shanqimike², WANG Hui², LI Guan¹, DENG Yu⁴

1.College of Life Sciences and Technology,Xinjiang University,Urumqi 830046,China;
2.Organisms Energy Research Institute of Xinjiang Academy of Agriculture Sciences,Urumqi 830091,China;
3.Research Institute of Food Crops,Xinjiang Academy of Agricultural Sciences, Urumqi 830091,China;
4.Biogas Research Institute of Ministry of Agriculture,Chengdu Sichuan,610041,China

Received:2017-11-16 Online:2017-12-25 Published:2018-01-29
Correspondence author: LI Guan(1949-),male,native place:Hubei,Professor,research field:Plant physiology,biochemistry and molecular biology. (E-mail) guanli@xju.edu.cn;DENG Yu (1963-),male,native place:Sichuan,Professor,research field:Biogas microorganism. (E-mail) dengyu@caas.cn
Supported by:
special funds for innovation and development for scientific and research institutes of Xinjiang Uygur Autonomous Region "Research on key recycling agricultural technology in the southern suburbs of Urumqi orogenic belt" (2016D04019) and Open fund for key laboratory of rural renewable energy development and utilization "Screening of salt tolerant cellulose degrading bacteria and optimization of fermentation process" (2017007); Xinjiang Academy of Agricultural Sciences Youth Fund Program "Biological pretreatment on biogas production of raw materials and gas production" (xjnky-2013021)

耐盐纤维素降解菌的筛选、鉴定及其配比优化的研究

王旭辉^1,2, 叶凯², 徐鑫³, 山其米克², 王卉², 李冠¹, 邓宇⁴

1.新疆大学生命科学与技术学院,乌鲁木齐 830046;
2.新疆农业科学院生物质能源研究所,乌鲁木齐 830091;
3.新疆农业科学院粮食作物研究所,乌鲁木齐 830091;
4.农业部沼气科学研究所,成都 610041

通讯作者: 李冠(1949-),男,湖北人,教授,博士生导师,研究方向为植物生理生化与分子生物学,(E-mail)guanli@xju.edu.cn;邓宇(1963-),男,四川人,教授,博士生导师,研究方向为沼气微生物,(E-mail)dengyu@caas.cn
作者简介:王旭辉(1985-),男,新疆人,助理研究员,博士研究生,研究方向为生物质能源利用,(E-mail)bio0407015@qq.com
基金资助:
新疆维吾尔自治区科研机构创新发展专项资金项目“乌鲁木齐南郊近山带循环农业关键技术研究”(2016D04019);农业部农村可再生能源开发利用重点实验室开放基金“耐盐纤维素降解菌的筛选及发酵工艺优化”(2017007);新疆农业科学院院青年基金项目“生物预处理对沼气生产原料及产气影响”(xjnky-2013021)

Abstract

Abstract: 【Objective】 To increase the degradation rate of maize straw by employing response surface method based on Box-Behnken (BBK).【Method】Four salt tolerant strains (WH2,WH4,WH7 and WH9) with good ability of degrading cellulose were isolated from saline alkali soil of corn straw manure. Based on morphologic observation,physiological and biochemical characteristics and molecular identification,they were identified as Bacillus cereus, Pseudomonas formosensis strain,Bacillus subtilis and Pseudomonas songnenensis respectively. On the basis of single-factor test analysis,Box-Behnken central composite design and response surface method (RSM),while taking the degradation rate of maize straw as the response value,response surface method was applied to analyze the influence of various factors and their interaction on the degradation rate of maize straws,thus optimizing the progress ratio of bacteria.【Result】The result showed that the optimum conditions were as follows:When the straw is 2.0 g,the inoculation quantity of the four strains (WH2,WH4,WH7,WH9) is 0.31 g, 0.21 g, 0.32 g and 0.22 g,respectively,and the rate of straw degradation reached the maximum.【Conclusion】Under these optimal conditions,the predictive value of the degradation rate of straw is 61.17%,meanwhile the verification value is 60.90%. There is only 0.27% difference between predictive and verification value,which has proved the rationality and effectiveness of RSM. The experiment has provided a theoretical basis for the microbial degradation of corn straws in Xinjiang.

Key words: corn straw; strains identification; degeneration rate; response surface methodology

摘要： 【目的】研究Box-Behnken(BBK)试验设计结合响应面分析法提高玉米秸秆的降解率。【方法】从玉米秸秆堆肥的盐碱土壤中分离筛选出4株高效耐盐纤维素降解菌WH2、WH4、WH7和WH9,经形态观察、生理生化反应和分子鉴定,确定4株菌分别为蜡样芽孢杆菌(Bacillus cereus)、荧光假单胞菌(Pseudomonas formosensis)、枯草芽孢杆菌(Bacillus subtilis)和松嫩假单胞菌(Pseudomonas songnenensis)。在单因素试验、BBK中心组合试验设计及响应面法分析法的基础上,以玉米秸秆降解率为响应值,利用响应面法研究各因素及其交互作用对玉米秸秆降解率的影响,优化菌种配比工艺。【结果】耐盐纤维素降解菌的最佳配比工艺:当秸秆添加量为2.0 g时,菌种WH2、WH4、WH7和WH9接种量分别为0.31、0.21、0.32和0.22 g时的玉米秸秆降解率达到最大值。【结论】在耐盐纤维素降解菌最优配比条件下,玉米秸秆降解率预测值为61.17%,验证值为60.90%,两值相差0.27%,证明了响应面法的合理性和有效性,为新疆盐碱地区玉米秸秆的微生物降解提供科学依据。

关键词: 玉米秸秆, 降解率, 菌种鉴定, 响应面法

CLC Number:

S188

WANG Xu-hui, YE Kai, XU Xin, Shanqimike, WANG Hui, LI Guan, DENG Yu. Study on the Isolation,Identification of Salt-tolerant Cellulose-Degrading Strains and Optimization of Its Strain Ratio[J]. Xinjiang Agricultural Sciences, 2017, 54(12): 2282-2292.

王旭辉, 叶凯, 徐鑫, 山其米克, 王卉, 李冠, 邓宇. 耐盐纤维素降解菌的筛选、鉴定及其配比优化的研究[J]. 新疆农业科学, 2017, 54(12): 2282-2292.

References 24

[1]	Rodrigues, A. C., Boroski, M., Shimada, N. S., Garcia, J. C., Nozaki, J., & Hioka, N. (2008). Treatment of paper pulp and paper mill wastewater by coagulation-flocculation followed by heterogeneous photocatalysis. Journal of Photochemistry & Photobiology A Chemistry, 194(1):1-10.
[2]	Georgakakis, D., & Krintas, T. (2000). Optimal use of the hosoya system in composting poultry manure. Bioresource Technology, 72(3): 227-233.
[3]	陈子爱,邓小晨. 微生物处理利用秸杆的研究进展 [J].中国沼气,2006,24(3):31-35.CHEN Zi-ai,DENG Xiao-chen. (2006). Progress in microbiologic utilization of crop straw [J]. China biogas,24(3):31-35. (in Chinese)
[4]	李平,王焰新,刘琨,等. 高效纤维素降解菌系的构建 [J].地球科学(中国地质大学学报),2009,(3):533-538.LI Ping,WANG Yan-xin,LIU Kun,et al. (2009). Construction of A Microbial System or Efficient Degradation of Cellulose [J]. China biogas,(3):533-538. (in Chinese)
[5]	Petersson, L., Kvien, I., & Oksman, K. (2007). Structure and thermal properties of poly(lactic acid)/cellulose whiskers nanocomposite materials. Composites Science & Technology, 67(11-12): 2,535-2,544.
[6]	李燕红,赵辅昆. 纤维素的研究进展 [J].生命科学,2005,17(5):392-397. LI Yan-hong,ZHAO Fu-kun. (2005). Research progress of cellulose [J]. Chinese Bulletin of Life Science,17(5):392-397. (in Chinese)
[7]	李争明,张娟,邓中洋,等. 纤维素酶产生菌的筛选、鉴定及发酵产酶条件优化 [J].生物技术通报,2015,31(5):146-152.LI Zheng-ming,ZHANG Juan,DENG Zhong-yang,et al. (2015). Screening,Identification and Optimization of Cellulase-producing Strains [J]. Biotechnology Bulletin,31(5):146-152. (in Chinese)
[8]	陈兴,王文元,汪显国,等. 烟叶纤维素降解菌的筛选、鉴定及其产酶条件优化 [J].云南大学学报,2015,37(2):323-328.CHEN Xing,WANG Wen-yuan,WANG Xian-guo,et al. (2015). Screening,identification of cellulose-decomposing strain from aging flue-cured tobacco leaves and optimization of its fermentation condition [J]. Journal of Yunnan University,37(2):323-328. (in Chinese)
[9]	张丽萍,李亚冰,程辉彩,等. 一株兼性厌氧纤维素酶产生菌的筛选、鉴定及其酶学性质研究 [J].华北农学报,2010,25(6):139-143.ZHANG Li-ping,LI Ya-bing,CHENG Hui-cai,et al. (2010). Screening and Identification of a Facultative Anaerobic Cellulase-producing Strain and Characterization of Cellulase [J]. Acta Agriculturae Boreali-Sinica,25(6):139-143. (in Chinese)
[10]	卢月霞,陈凯,李海江.一株纤维素降解细菌的筛选及产酶条件研究 [J].安徽农业科学,2007,35(12):3 631-3 644.LU Yue-xia,CHEN Kai,LI Hai-jiang. (2007). Screening of Cellulose-degrading Bacteria and Study on its Cellulose producing Condition [J]. Journal of Anhui Agricultural Sciences,35(12):3,631-3,644. (in Chinese)
[11]	Haruta, S., Cui, Z., Huang, Z., Li, M., Ishii, M., & Igarashi, Y. (2002). Construction of a stable microbial community with high cellulose-degradation ability. Applied Microbiology & Biotechnology, 59(4-5): 529-534.
[12]	吴俊妹,马安周,崔萌萌,等.降解纤维素产甲烷的四菌复合系 [J].环境科学,2014,35(1):327-333.WU Jun-mei,MA An-zhou,CUI Meng-meng,et al. (2014). Screening of Cellulose-degrading Bacteria and Study on its Cellulose producing Condition [J]. Environmental Science,35(1):327-333. (in Chinese)
[13]	王旭辉,晁群芳,徐鑫,等. 高效石油降解菌的筛选、鉴定及其配比优化的研究 [J].工业安全与环保,2013,2(10):42-46.WANG Xu-hui,CHAO Qun-fang,XU Xin,et al. (2013). Bioconversion of Cellulose to Methane by a Consortium Consisting of Four Microbial Strains [J]. Industrial Safety and Environmental Protection,2(10):42-46.(in Chinese)
[14]	孔雅丽,贾辉,王生荣,等.一株东祁连山耐低温纤维素降解菌株的分离、鉴定及产酶特性[J].工业微生物,2014,44 (1) :60-66.KONG Ya-li,JIA Hui,WANG Sheng-rong,et al. (2014). Isolation,identification and characterization of a cold-adapted cellulose degrading strain from eastern Qilian mountains industrial microbiology [J]. Industrial Microbiology,44 (1): 60-66(in Chinese)
[15]	王平宇,张树华.硅酸盐细菌的分离及生理生化特性的鉴定 [J].南昌航空工业学院学报,2001,15(2):78-81.WANG Ping-yu,ZHANG Shu-hua.(2001). Study on the Isolation Sillicate Bacteria and Its Identification of Biochemical Characteristics [J]. Journal of Nanchang Institute of Aeronautical Technology,15 (2):78-81.(in Chinese)
[16]	东秀珠,蔡妙英.常见细菌鉴定手册 [K].北京:科学出版社,2001.DONG Xiu-zhu,CAI Miao-ying. (2001). Common bacterial identification manual [K]. Beijing: Science Press.(in Chinese)
[17]	王玉万,徐文玉. 木质纤维素固体基质发酵物中半纤维素、纤维素和木素的定量分析程序 [J].微生物学通报,1987,14 (2):35-38.WANG Yu-wan,XU Wen-yu. (1987). Quantitative Analysis of Hemicellulose,Cellulose and Lignin in Lignocellulose Solid Substrate Fermentation [J]. Microbiology China,14 (2):35-38. (in Chinese)
[18]	李华,孔新刚,王俊. 秸秆饲料中纤维素、半纤维素和木质素的定量分析研究 [J]. 新疆农业大学学报,2007,30 (3):65-68.LI Hua,KONG Xin-gang,WANG Jun. (2007). Study on Quantitative Analysis of Hemicellulose and Cellulose and Lignin in Roughage of Cereal Straw [J]. Journal of Xinjiang Agricultural University,30(3): 65-68. (in Chinese)
[19]	薛惠琴,杭怡琼,陈谊.稻草秸秆中木质素、纤维素测定方法的研讨[J]. 上海畜牧兽医通讯,2001,(2):15.XUE Hui-qin,HANG Yi-qiong,CHEN Yi. (2001). Study on Determination Method of Lignin and Cellulose in Straw [J]. Shanghai Journal of Animal Husbandry and Veterinary Medicine,(2):15. (in Chinese)
[20]	Box, G. E. P., & Behnken, D. W. (1960). Some new three level designs for the study of quantitative variables. Technometrics, 2(4): 455-475.
[21]	Annadurai, G., & Sheeja, R. Y. (1998). Use of box-behnken design of experiments for the adsorption of verofix red using biopolymer. Bioprocess Engineering, 18(6): 463-466.
[22]	林标声,张彭湃,杨生玉,等. 响应面分析法优化丙酮酸发酵中维生素水平 [J].食品科技,2007,(4):36-39.LIN Biao-sheng,ZHANG Peng-pai,YANG Sheng-yu,et al. (2007). Application of Response Surface method to optimization of vitamin in levels on the fermentation of pyruvic acid [J]. Food Science and Technology,(4):36-39.(in Chinese)
[23]	刘晓梅,邹亚杰,胡清秀,等. 菌渣纤维素降解菌的筛选与鉴定 [J].农业环境科学学报,2015,34(7):1 384-1 391.LIU Xiao-mei,ZOU Ya-jie,HU Qing-xiu,et al. (2015). Screening and Identification of Cellulose-Degrading Bacteria from Spent Substrate of Edible Mushroom [J]. Journal of Agro-Environment Sciencey,34(7): 1,384-1,391.(in Chinese)
[24]	冯忻,裴宇航,周晓飞,等. 纤维素降解菌的筛选与高效混合菌群的构建 [J].西北农林科技大学学报,2012,25 (4) :30-32.FENG Xin,PEI Yu-hang,ZHOU Xiao-fei,et al. (2012). Screening of stalk cellulose-degrading microorganism and construction of microbial community with high capacity [J]. Journal of Northwest A & F University (Natural Science Edition) ,25(4):30-32. (in Chinese)

Study on the Isolation,Identification of Salt-tolerant Cellulose-Degrading Strains and Optimization of Its Strain Ratio

耐盐纤维素降解菌的筛选、鉴定及其配比优化的研究

PDF

Knowledge

Abstract

Cite this article

share this article

References 24

Related Articles 15

Recommended Articles

Metrics

[1]	ZHAO Jinghua, YANG Tingrui, ZHANG Heng, Hudan Tumaibai, MA Liang, CHEN Kaili. Optimal Selection of Water and Fertilizer for Spring Wheat under Drip Irrigation in Gravel Sandy Soil Based on Response Surface Methodology [J]. Xinjiang Agricultural Sciences, 2023, 60(1): 43-51.
[2]	LEI Jing, BI Jingfeng, LIAN Weijia, CHEN Ya, HAN Chen. Optimization of Pulsed Sudden Decompression Flash Drying Process for Crispy Raisin by Response Surface Methodology [J]. Xinjiang Agricultural Sciences, 2021, 58(8): 1529-1539.
[3]	LEI Jing, CHEN Ya, LIAN Weijia, HAN Chen, Ayijiamali Jiapaer, REN Hongsong. Optimization of Fermentation Technology for Mulberry Fruit and Leaf Wine by Response Surface Methodology [J]. Xinjiang Agricultural Sciences, 2021, 58(6): 1113-1123.
[4]	Xie Yuqing, DAI Jinping, Chen Jin, Wang Zhifang, GuliAhmat, Yang Xinping, Zhang Huitao, Wang Xiaowu, Feng Lei. Optimizing Carbon and Nitrogen Source in Liquid Fermentation Medium of G.resinaceum LZ02 Producing Laccase with Response Surface Methodology [J]. Xinjiang Agricultural Sciences, 2020, 57(5): 869-876.
[5]	Xie Hui , Wang Xian , Zhang Wen , Wu Guo-hong , Wang Min , Han Shou-an , Yi Zhati·Tuerdahong , Zhong Hai-xia , Pan Ming-qi , Lu Sheng-zu ,Li Jiang. Polyphenols from Raisins in Xinjiang: Optimization of Ultrasonic-assisted Extraction Process by Response Surface Methodology [J]. Xinjiang Agricultural Sciences, 2019, 56(4): 696-706.
[6]	WANG Xu-hui^1,2, XU Xin ³, WANG Hui², YE Kai², TU Zhen-dong², CHAO Qun-fang¹. Optimization, Purifcation and Preliminary Structural Characterization of Artemisia rupestris L. Polysaccharides [J]. Xinjiang Agricultural Sciences, 2018, 55(6): 1085-1097.
[7]	XIE Jing-long, LI Xiao-bin, ZHAO Fang, WANG Chen-chen, YANG Kai-lun. Optimization of Fermentation Medium Components for Dihydrodaidzein Producing Strain Using RSM [J]. Xinjiang Agricultural Sciences, 2018, 55(1): 143-155.
[8]	XIE Yu-mei;LIU Fang-hui;DING Xin-hua;LI Cui-mei;FU Kai-yun;HE Jiang;Tursun Ahmat;Abdureyim Abdurexit;LI Peng-fa;XU Jian-jun;GUO Wen-chao. Straw Storage Effects on the Pupation and Eclosion of Overwintering Larvae of Corn Borer [J]. , 2017, 54(3): 505-512.
[9]	XU Xin, MAO Hong-yan, YU Ming. Study on Optimization of Chickpea Resistant Starch Preparation and Its Structural Properties [J]. Xinjiang Agricultural Sciences, 2017, 54(10): 1847-1855.
[10]	. Study on the Optimization of Fermentation Conditions and Stability of Melanin from a Radiation-resistant Aureobasidium pullulans [J]. , 2016, 53(9): 1692-1699.
[11]	XU Xin;YU Ming;MAO Hong-yan;WU Xin-feng. Study on Preparation Technology of the Chickpea Resistant Starch by Autoclaving Treatment [J]. , 2016, 53(12): 2250-2257.
[12]	WANG Xu-hui;ZHAO Xiao-mei;XU Xin;YE Kai;WANG Hui. Research on Optimization of the Extraction Method of Pomegranate Peel by Using Response Surface Methodology [J]. , 2015, 52(10): 1801-1807.
[13]	MO Hai-yan;BAO Hui-fang;WANG Ning;QI Hong-liang;WANG Wei. Optimization of Fermentation Medium for Enterococcus faecalis by Response Surface Method [J]. , 2014, 51(8): 1532-1540.
[14]	CAO Jing;BAI Yu-jia;CAO Yan;ZHENG Jie;DUAN Ji-hua;FENG Zuo-shan. Optimization of Microwave Extraction Process for Hop Stems by Using Response Surface Methodology [J]. , 2014, 51(10): 1869-1877.
[15]	QIN Xin-zheng;YANG Xin-ping;CHEN Jing;FENG Lei. Optimization of Incubated Mycelium of Mortierella alpine Based on Response Surface Methodology [J]. , 2014, 51(1): 89-97.