Screening and Identification of a Cellulose Degrading Strain

Yang Na, He Xin, Du Chunmei

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Chinese Agricultural Science Bulletin ›› 2021, Vol. 37 ›› Issue (17) : 26-31. DOI: 10.11924/j.issn.1000-6850.casb2021-0131

Screening and Identification of a Cellulose Degrading Strain

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Abstract

To solve the problem of low degradation rate of corn straw, stable cellulose degrading bacteria were screened from natural fermentation compost with amino acid tail liquor as the nitrogen source, aiming to lay a foundation for the preparation of efficient straw decomposition agent. Congo red staining method was adopted for preliminary screening, the determination of cellulase activity and the loss rate of corn straw weight were used for re-screening, and morphological and molecular biological methods were used to identify the efficient degrading strain. The results showed that an efficient cellulose degrading strain, SC2, was obtained. The filter paper enzyme activity, endoglucanase activity, exoglucanase activity and β-glucosidase activity of SC2 was 17.70 U/mL, 58.97 U/mL, 16.85 U/mL and 79.26 U/mL, respectively. The degradation rate of corn straw reached 33.07%. According to the colony characteristics, sporulation structure, spore morphology and ITS sequence, SC2 was identified as Aspergillus niger. Strain SC2 has good cellulose degradation ability, can effectively promote the degradation of straw and be used to prepare corn straw decomposing agent.

Key words

corn straw / corn straw cellulose / cellulose degradation / degradation / enzyme activity / screening / identification

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Yang Na , He Xin , Du Chunmei. Screening and Identification of a Cellulose Degrading Strain. Chinese Agricultural Science Bulletin. 2021, 37(17): 26-31 https://doi.org/10.11924/j.issn.1000-6850.casb2021-0131

References

[1]
Li F H, Ding Z T, Ke W C, et al. Ferulic acid esterase-producing lactic acid bacteria and cellulase pretreatments of corn stalk silage at two different temperatures: Ensiling characteristics, carbohydrates composition and enzymatic saccharification[J]. Bioresource Technology, 2019,282:211-221.
[2]
Singh J, Suhag M, Dhaka A. Augmented digestion of lignocellulose by steam explosion, acid and alkaline pretreatment methods: A review[J]. Carbohydr Polym, 2015,117:624-631.
[3]
Ma L L, Lu Y Y, Yan H, et al. Screening of cellulolytic bacteria from rotten wood of Qinling (China) for biomass degradation and cloning of cellulases from Bacillus methylotrophicus[J]. BMC Biotechnology, 2020,20(6):1-13.
[4]
Shakoor S, Aftab S, Rehman A. Characterization of cellulose degrading bacterium, Bacillus megaterium S3, isolated from indigenous environment[J]. Pakistan Journal of Zoology, 2013,45(6):1655-1662.
[5]
Krishna M, Guder D G. Isolation and characterization of potential cellulose degrading bacteria from sheep rumen[J]. Journal of Pure and Applied Microbiology, 2019,13(3):1831-1839.
[6]
Sun S N, Sun S L, Cao X F, et al. The role of pretreatment in improving the enzymatic hydrolysis of lignocellulosic materials[J]. Bioresource Technology, 2016,199:49-58.
[7]
Carlos M, Bruna S, Betsy M, et al. A metagenomic advance for the cloning and characterization of a cellulase from red rice crop residues[J]. Molecules, 2016,21(7):1-12.
[8]
宋文芳, 胡国全, 徐彦胜. 一株低温厌氧纤维素降解细菌的分离、鉴定及其酶学性质的研究[J]. 西南农业学报, 2011,24(4):1317-1322.
[9]
Bansal N, Tewarir R, Soni R, et al. Production of cellulases from Aspergillus niger NS-2 in solid state fermentation on agricultural and kitchen waste residues[J]. Waste Management, 2012,32(7):1341-1346.
[10]
Egwuatu T F, Appeh O G. Isolation and characterization of filter paper degrading bacteria from the guts of coptotermes formosanus[J]. Journal of Bioremediation & Biodegradation, 2018,9(3):1-5.
[11]
Milala M A, Shugaba A, Gidado A, et al. Studies on the use of agricultural wastes for cellulase enzyme production by Aspegillus niger[J]. Research Journal of Agriculture and Biological Sciences, 2005,1(4):325-328.
[12]
Schwarz W H. The cellulosome and cellulose degradation by anaerobic bacteria[J]. Applied Microbiology & Biotechnology, 2001,56(5-6):634-649.
[13]
Rastogi G, Muooidi G L, Gurram R N, et al. Isolation and characterization of cellulose-degrading bacteria from the deep subsurface of the Homestake gold mine, Lead, South Dakota, USA[J]. Journal of Industrial Microbiology & Biotechnology, 2009,36(4):585-598.
[14]
Karthika A, Seenivasagan R, Kasimani R, et al. Cellulolytic bacteria isolation, screening and optimization of enzyme production from vermicompost of paper cup waste[J]. Waste Management, 2020,116:58-65.
[15]
Farjana I, Naranay R. Screening, purification and characterization of cellulase from cellulase producing bacteria in molasses[J]. BMC Research Notes, 2018,11(1):445-451.
This study was conducted to isolate, screening and purification of cellulase from bacteria present in sugar industry waste (molasses) and characterization by morphological and biochemical analysis.Based on experiments, three bacterial strains produced clear transparent zone into carboxymethyl cellulose (CMC) agar plate were identified as cellulase producing bacteria. Different culture parameters such as pH, temperature, incubation period, substrate concentration and carbon sources were optimized for enzyme production. According to the morphological and biochemical tests, the isolated strains were identified as Paenibacillus sp., Bacillus sp. and Aeromonas sp. The first strain Paenibacillus sp. showed high potentiality for maximum cellulase production (0.9 µmol ml min) at pH 7.0 after 24 h of incubation at 40 °C in a medium containing 1.0% CMC. Then Paenibacillus sp. was selected for enzyme purification by ammonium sulfate precipitation, DEAE-cellulose and CM-cellulose column chromatography, respectively. In last step of purification, specific activity, recovery and purification fold were 2655 U/mg, 35.7% and 9.7, respectively. The molecular weight of the purified cellulase was found to be 67 kDa by SDS-PAGE, had an optimal pH and temperature at 7.0 and 40 °C. According to substrate specificity, the purified cellulase had high specificity on CMC substrate which indicated it to be an endo-β-1,4-glucanase.
[16]
张爱梅, 殷一然, 齐汝楠. 产纤维素酶沙棘根瘤内生放线菌的筛选,鉴定及其酶活性测定[J]. 西北师范大学学报:自然科学版, 2019,208(5):75-80,96.
[17]
刘晓飞, 宋洁, 马京求, 等. 产纤维素酶放线菌的筛选鉴定及其对玉米秸秆的降解[J]. 精细化工, 2020,37(8):1657-1664,1671.
[18]
张健, 李燕婷, 袁亮, 等. 氨基酸发酵尾液可促进樱桃番茄对水溶肥料氮素的吸收利用[J]. 植物营养与肥料学报, 2018,24(1):114-121.
[19]
陈莉. 绿色木霉固态发酵产纤维素酶条件研究[J]. 安徽农业科学, 2013,41(7):2823-2825.
[20]
李晓秀, 张盼, 刘琬瑜, 等. 玉米秸秆降解真菌的筛选及鉴定[J]. 饲料工业, 2017,38(3):37-42.
[21]
陈建爱, 贾梦莹, 陈为京. PDA中水含量对固体培养黄绿木霉T1010分生孢子生长发育的影响[J]. 中国农学通报, 2020,558(15):112-120.
[22]
李正风, 李岩, 朱杰, 等. 烟草秸秆中产纤维素酶细菌筛选,鉴定及酶活测定[J]. 西南农业学报, 2020,33(3):645-650.
[23]
许玉林, 郑月霞, 叶冰莹, 等. 一株纤维素降解真菌的筛选及鉴定[J]. 微生物学通报, 2013,40(2):220-227.
[24]
Azhar A H, Mohamed S A, Hoda H A, et al. Optimization and molecular identification of novel cellulose degrading bacteria isolated from Egyptian environment[J]. Journal of Genetic Engineering and Biotechnology, 2017,15:77-85.
[25]
Gupta P, Samant K, Sahu A. Isolation of cellulose-degrading bacteria and determination of their cellulolytic potential[J]. International Journal of Microbiology, 2012,2012(6):1-5.
[26]
黄亚丽, 黄媛媛, 马慧媛, 等. 低温秸秆降解真菌的筛选及在秸秆还田中的应用[J]. 中国农学通报, 2020,36(21):53-60.
[27]
Sun Y, Qu J B, Li R L, et al. Optimization of the enzyme production conditions of Bacillus licheniformis and its effect on the degradation of corn straw[J]. Journal of Biobased Materials and Bioenergy, 2018,12(5):432-440.
[28]
张冬雪, 文亚雄, 罗志威, 等. 纤维素降解菌的分离筛选及其对水稻秸秆的降解效果分析[J]. 江西农业学报, 2020,32(1):76-80.
[29]
王勇, 张育铭, 朱洪磊, 等. 高效纤维素降解菌的筛选及产酶活力测定[J]. 江苏农业科学, 2020,48(23):255-260.
[30]
孙美娜, 张凡凡, 王永强, 等. 棉花秸秆纤维素降解菌的筛选鉴定与降解棉秆效果研究[J]. 新疆农业科学, 2018,55(1):16-23.
[31]
李思杨. 不同氮源和菌剂处理对水稻秸秆腐解规律及土壤养分的影响[D]. 沈阳:沈阳农业大学, 2019.
[32]
Gong X, Zou H, Qian C, et al. Construction of in situ degradation bacteria of corn straw and analysis of its degradation efficiency[J]. Annals of Microbiology, 2020,70(1):1-15.
[33]
李静, 张瀚能, 赵翀, 等. 高效纤维素降解菌分离筛选、复合菌系构建及秸秆降解效果分析[J]. 应用与环境生物学报, 2016,22(4):689-696.

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Copyright reserved © 2021. Chinese Agricultural Association Bulletin. All articles published represent the opinions of the authors, and do not reflect the official policy of the Chinese Agricultural Association or the Editorial Board, unless this is clearly specified.
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