纤维素酶产生菌的筛选、鉴定及发酵产酶条件优化

doi:10.13560/j.cnki.biotech.bull.1985.2015.05.023

生物技术通报 ›› 2015, Vol. 31 ›› Issue (5): 146-152.doi: 10.13560/j.cnki.biotech.bull.1985.2015.05.023

纤维素酶产生菌的筛选、鉴定及发酵产酶条件优化

李争明¹, 张娟¹, 邓中洋¹, 卢凡¹, 秦文胜^2,

（1.湖北工业大学资源与环境工程学院微藻实验室工业发酵湖北省协同创新中心河湖生态修复及藻类利用湖北省重点实验室，武汉 430068；2. Department of Biology，Lakehead University，Ontario，P7B 5E1，Canada）

收稿日期:2014-09-10 出版日期:2015-05-18 发布日期:2015-05-18
作者简介:李争明，男，硕士研究生，研究方向：生物化工；E-mail：lizm1987@163.com
基金资助:
湖北工业大学·楚天学者启动基金项目

Screening，Identification and Optimization of Cellulase-producing Strains

Li Zhengming¹, Zhang, Juan¹, Deng Zhongyang¹, Lu Fan¹, Qin Wensheng²

（1. Microalgae Laboratory，School of Resource & Environmental Engineering，Hubei Provincial Cooperative Innovation Center of Industrial Fermentation，Hubei Provincial Key Laboratory of Lake Ecological Restoration & Algae Utilization，Hubei University of Technology，Wuhan 430068；2. Department of Biology，Lakehead University，Ontario，P7B 5E1，Canada）

Received:2014-09-10 Published:2015-05-18 Online:2015-05-18

摘要/Abstract

摘要： 以腐烂木材、腐殖土等材料作为菌源，从中分离出180个菌株，以期得到具有纤维素酶活性的菌株。采用革兰氏碘液染色法进行定性初筛，获得了44个纤维素酶产生菌株。将此44个菌株发酵培养后，使用滤纸酶活性测定法进行定量复筛，滤纸酶活性最高的是菌株J1-3-1。通过对J1-3-1菌株的16S rRNA 的序列测定分析，将J1-3-1鉴定为鞘氨醇杆菌（Sphingobacterium sp.）。经对J1-3-1进行产酶发酵条件优化，分别确立了产生最高滤纸酶（FPase）活性、内切葡聚糖酶（CMCase）活性和β-葡萄糖苷酶（β-glucosidase）活性的发酵条件。在最优发酵产酶条件下，菌株J1-3-1的滤纸酶、内切葡聚糖酶和β-葡萄糖苷酶的最高酶活性分别为8.76、28.04和7.02 U/mL。

关键词: 纤维素酶产生菌, 筛选, 鉴定, 发酵, 鞘氨醇杆菌

Abstract: To isolate efficient cellulase-producing bacteria, 180 bacterial isolated from rotten wood, humus, and other soil samples, and 44 were screened from them and confirmed as cellulase-producing bacteria by Gram’s iodine solution staining. In the subsequent secondary screening tests, those 44 isolates were cultured in fermentation media and their total cellulase activities（FPase）were measured and compared. The strain J1-3-1, with the highest FPase activity, was identified as Sphingobacterium sp. by 16S rRNA sequence analysis. The optimal fermentation conditions for enzymes production of J1-3-1 were determined. Under the optimal conditions, its activities of FPase, CMCase, and β-glucosidase reached 8.76, 28.04和7.02 U/mL, respectively. The results demonstrated that J1-3-1 was a promising candidate for potential industrial production of cellulase.

Key words: cellulase-producing bacteria, screening, identification, fermentation, Sphingobacterium sp.

李争明, 张娟, 邓中洋, 卢凡, 秦文胜. 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 Zhengming, Zhang, Juan, Deng Zhongyang, Lu Fan, Qin Wensheng. Screening，Identification and Optimization of Cellulase-producing Strains[J]. Biotechnology Bulletin, 2015, 31(5): 146-152.

参考文献

[1] Agarwal AK. Biofuels（alcohols and biodiesel）applications as fuels for internal combustion engines[J]. Progress in Energy and Combustion Science, 2007, 33：233-271.
[2] Escobar JC, Lora ES, Venturini OJ, et al. Biofuels：environment, technology and food security[J]. Renewable and Sustainable Energy Reviews, 2009, 13：1275-1287.
[3] Singh A, Pant D, Korres NE, et al. Key issues in life cycle assessment of ethanol production from lignocellulosic biomass：challenges and perspectives[J]. Bioresource Technology, 2010, 101（13）：5003-5012.
[4] Prasad S, Singh A, Jain N, et al. Ethanol production from sweet sorghum syrup for utilization as automotive fuel in India[J]. Energy & Fuel, 2007, 21（4）：2415-2420.
[5] Singh A, Smyth BM, Murphy JD. A biofuel strategy for Ireland with an emphasis on production of biomethane and minimization of land-take[J]. Renewable and Sustaiable Energy Reviews, 2010, 14（1）：277-288.
[6] Prasad S, Singh A, Joshi HC. Ethanol as an alternative fuel from agricultural, industrial and urban residues[J]. Resource, Conservation and Recycling, 2007, 50：1-39.
[7] Himmel ME, Ding SY, Johnson DK, et al. Enzymes for biofuels production biomass recalcitrance：Engineering plants and enzymes for biofuels production[J]. Science, 2007, 315（5813）：804-807.
[8] Ding SY, Xu Q, Crowley M, et al. A biophysical perspective on the cellulosome：new opportunities for biomass conversion[J]. Current Opinion in Biotechnology, 2008, 19（3）：218-227.
[9] Maki M, Leung KT, Qin WS. The prospects of cellulase-producing bacteria for the bioconversion of lignocellulosic biomass[J]. International Journal of Biological Sciences, 2009, 5（5）：500-516.
[10] Kasana RC, Salwan R, Dhar H, et al. A rapid and easy method for the detection of microbial cellulases on agar plates using gram’s iodine[J]. Current Microbiology, 2008, 57：503-507.
[11] Maki ML, Broere M, Leung KT, et al. Characterization of some efficient cellulase producing bacteria isolated from paper mill sludges and organic fertilizers[J]. International Journal of Biochemistry Molecular Biology, 2011, 2：146-154.
[12] Zhang YHP, Hong J, Ye XH. Cellulase assays[J]. Biofuels：Methods in Molecular Biology, 2009, 581：213-231.
[13] Ghose TK. Measurement of cellulase activities[J]. International Union of Pure and Applied Chemistry, 1987, 59（2）：257-268.
[14] 武峥, 张迎君, 周心智. 降解秸秆的纤维素酶产生菌的筛选及产酶条件研究[J]. 纤维素科学与技术, 2009, 17（2）：20-26.
[15] 包衎, 王晓辉, 张伟琼, 等. 纤维素分解菌的选育和酶活测定[J]. 生物学杂志, 2007, 24（2）：56-58.
[16] Vandamme P, Pot B, Gills M, et al. Polyphasic taxonomy, a consensus approach to bacterial systematics[J]. Microbiology Review, 1996, 60（2）：407-438.
[17] Wei GF, Pan L, Du HM, et al. ERIC-PCR fingerprinting-based community DNA hybridization to pinpoint genome specific fragments as molecular markers to identify and track populations common to healthy human guts[J]. Journal of Microbial Methods, 2004, 59（1）：91-108.
[18] 高永超, 王加宁, 孔学, 等. 石油降解菌多食鞘氨醇杆菌的发酵条件优化[J]. 生物技术, 2009, 19（1）：74-77.
[19] Yu Y, Zhang W, Chen GH, et al. Preparation of petroleum-degrading bacterial agent and its application in remediation of contaminated soil in Shengli Oil Field, China[J]. Environmental Science and Pollution Research, 2014, 21：7929-7937.
[20] 董至恒, 孙东阳, 高焕, 等. 一株转化井冈霉素产生井冈霉胺细菌的筛选与鉴定[J]. 中国抗生素杂志, 2009, 34（2）：79-82.
[21] 王飞, 张丽. 超声波破碎法提取多食鞘氨醇杆菌中17β-雌二醇降解酶的优化及酶学性质[J]. 江苏农业科学, 2014, 42（4）：310-313.
[22] Ariffin H, Abdullah N, Kalsom MS, et al. Production and characterization of cellulase by Bacillus pumilus EB3[J]. International Journal of Engineering and Technology, 2006, 3（1）：47-53.

纤维素酶产生菌的筛选、鉴定及发酵产酶条件优化

Screening，Identification and Optimization of Cellulase-producing Strains

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