产高温纤维素酶木霉菌株的筛选、鉴定及酶学性质研究

中国生物工程杂志

2012, Vol. 32

Issue (07): 60-65

研究报告

产高温纤维素酶木霉菌株的筛选、鉴定及酶学性质研究

杨捷^1,2, 叶秀云^1,2, 严芬¹, 高珍娜¹, 李仁宽^1,2, 吕文静¹, 林娟^1,2

1. 福州大学生物科学与工程学院福州 350108;
2. 酶高效表达国家工程实验室福州 350002

Screening and Classification of a Thermophilic Cellulase-producing Trichoderma sp. Strain and the Study of the Enzyme Properties

YANG Jie^1,2, YE Xiu-yun^1,2, YAN Fen¹, GAO Zhen-na¹, LI Ren-kuan^1,2, LV Wen-jing¹, LIN Juan^1,2

1. College of Biological Sciences and Technology, Fuzhou University, Fuzhou 350108, China;
2. National Engineering Laboratory for Enzyme Expression, Fuzhou 350002, China

全文: PDF(735 KB) HTML

摘要： 从稻草堆肥中筛选得到一株产高温纤维素酶的霉菌M1,通过形态学观察和分子生物学鉴定,确定其为木霉属(Trichoderma)。在稻草液体发酵培养基中,木霉M1的CMC酶(carboxymethyl cellulase, CMCase)合成模式为同步合成型。酶学性质研究表明,此CMC酶的最适反应pH为4.4,在pH 4.0~6.0保温4h仍可保持95%以上的酶活力;其最适反应温度为75℃,在50℃下保温4h,可保持87%的酶活力;60℃下保温4h,可保持65%的酶活力,具有较好的热稳定性。

关键词： 木霉; CMC酶; 高温; 筛选; 鉴定; 酶学性质

Abstract: A fungal strain, designated as M1, producing thermo-tolerant cellulase was isolated from straw compost and classified as Trichoderma sp. via morphological analysis and its 18S rDNA sequence. The pattern of cellulase synthesis by the strain in submerged fermentation with rice straw as the sole carbon source was investigated as well as the enzyme properties of the carboxymethyl cellulase (CMCase). The maximum CMCase activity was observed at pH 4.4 and the CMCase retained more than 95% activity in the pH range of 4.0~6.0 after incubation at 50℃ for 4h. The optimum temperature for the CMCase activity was 75℃; this CMCase maintained 87% and 65% of the initial activity after incubation for 4h at 50℃ and 60℃, respectively, indicating that the enzyme had good thermostability.

Key words: Trichoderma sp. CMCase Thermophilic Screening Classification Enzyme properties

收稿日期: 2012-02-01 出版日期: 2012-07-25

ZTFLH:

Q55

基金资助: 福建省科技厅项目(2010J05073);福建省高校创新团队培育计划资助项目

通讯作者: 林娟 E-mail: ljuan@fzu.edu.cn

	服务
	把本文推荐给朋友
	加入引用管理器
	E-mail Alert
	RSS
	作者相关文章
	杨捷
	叶秀云
	严芬
	高珍娜
	李仁宽
	吕文静
	林娟

引用本文:

杨捷, 叶秀云, 严芬, 高珍娜, 李仁宽, 吕文静, 林娟. 产高温纤维素酶木霉菌株的筛选、鉴定及酶学性质研究[J]. 中国生物工程杂志, 2012, 32(07): 60-65.

YANG Jie, YE Xiu-yun, YAN Fen, GAO Zhen-na, LI Ren-kuan, LV Wen-jing, LIN Juan. Screening and Classification of a Thermophilic Cellulase-producing Trichoderma sp. Strain and the Study of the Enzyme Properties. China Biotechnology, 2012, 32(07): 60-65.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/ 或 https://manu60.magtech.com.cn/biotech/CN/Y2012/V32/I07/60

[1] Kubicek C P, Messner R, Gruber F, et al. The Trichoderma cellulase regulatory puzzle:From the interior life of a secretary fungus. Enzyme Microb Technol, 1993, 15 (2): 90-99.
[2] Zhang Y H P, Himmel M E, Mielenz J R. Outlook for cellulase improvement: screening and selection strategies. Biotechnol Adv, 2006, 24 (5): 452-481.
[3] Freier D, Mothershed C P, Wiegel J. Characterization of Clostridium thermocellum JW20. Appl Environ Microbiol, 1988, 54 (1): 204-211.
[4] Bronnenmeier K, Kern A, Liebl W, et al. Purification of Thermotoga maritima enzymes for the degradation of cellulose materials. Appl Environ Microbiol, 1995, 61 (4): 1399-1407.
[5] Yang S J, Kataeva I, Hamilton-Brehm S D, et al. Efficient degradation of lignocellulosic plant biomass without pretreatment by the thermophilic anaerobe, Anaerocellum thermophilum DSM 6725. Appl Environ Microbiol, 2009, 75 (14): 4762-4769.
[6] Alani F, Anderson W A, Moo-Young M. New isolate of Streptomyces sp. with novel thermoalkalotolerant cellulases. Biotechnol Lett, 2008, 30 (1): 123-126.
[7] Wojtczak G, Breuil C, Yamada J, et al. A comparison of the thermostability of cellulases from various thermophilic fungi. Appl Environ Microbiol, 1987, 27 (1): 82-87.
[8] Liu S, Duan X, Lu X, et al. A novel thermophilic endoglucanase from a mesophilic fungus Fusarium oxysporum. Chin Sci Bull, 2006, 51 (2): 191-197.
[9] Li D C, Li A N, Papageorgiou A C. Cellulases from thermophilic fungi: recent insights and biotechnological potential. Enzyme Res, 2011, 2011: 308730.
[10] Heinzelman P, Snow C D, Wu I, et al. A family of thermostable fungal cellulases created by structure-guided recombination. PNAS, 2009, 106 (14): 5610-5615.
[11] Hendricks C W, Doyle J D, Hugley B. A new solid medium for enumerating cellulose-utilizing bacteria in soil. Appl Environ Microbiol, 1995, 61 (5): 2016-2019.
[12] 魏景超. 真菌鉴定手册. 上海: 上海科学技术出版社, 1979: 487-494. Wei J C. Manual of Fungal Characterization. Shanghai: Shanghai Scientific and Technical Publishers. 1979: 487-494.
[13] 杨民和, 徐焰平, 苏经迁. 茶树内生球座菌的鉴定及遗传多样性分析. 江西农业大学学报, 2010, 32 (5): 946-955. Yang M H, Xu Y P, Su J Q. Characterization and genetic diversity analysis of endophytic Guignardia isolated from Camellia sinensis. Acta Agric Univ Jiangxiensis, 2010, 32 (5): 946-955.
[14] Thompson J D, Gibson T J, Plewniak F, et al. The Clustal_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tool. Nucleic Acids Res, 1997, 25 (24): 4876-4882.
[15] Kumar S, Tamura K, Nei M. MEGA3: integrated software for molecular evolutionary genetics analysis and sequence alignment. Brief Bioinform, 2004, 5 (2): 150-163.
[16] Liu G, Xu Z, Cen P. A morphologically structured model for mycelial growth and secondary metabolite formation. Chin J Chem Eng, 2000, 8 (1): 46-51.
[17] Mandels M, Andreotti R, Roche C. Measurement of saccharifying cellulase. Biotechnol Bioeng Symp, 1976, 6: 21-33.

[1]	郭芳,张良,冯旭东,李春. 植物源UDP-糖基转移酶及其分子改造^*[J]. 中国生物工程杂志, 2021, 41(9): 78-91.
[2]	钱昱,丁晓雨,刘志强,袁增强. 基因修饰人多能干细胞的高效单克隆建系方法[J]. 中国生物工程杂志, 2021, 41(8): 33-41.
[3]	张虎,刘镇洲,陈家敏,高保燕,张成武. 利用海洋硅藻生产生物活性物质研究进展^*[J]. 中国生物工程杂志, 2021, 41(4): 81-90.
[4]	栗波,王泽建,梁剑光,刘爱军,李海东. 等离子体作用结合氧限制模型选育利福霉素SV高产菌株 ^*[J]. 中国生物工程杂志, 2021, 41(2/3): 38-44.
[5]	魏子翔,张柳群,雷磊,韩正刚,杨江科. *疏棉状嗜热丝孢菌(Thermomyces lanuginosus)脂肪酶的理性设计提高其活性和温度稳定性*[J]. 中国生物工程杂志, 2021, 41(2/3): 63-69.
[6]	邓蕊,曾佳利,卢雪梅. 基于Musca domestica cecropin的抗肿瘤小分子衍生肽筛选及构效关系解析^*[J]. 中国生物工程杂志, 2021, 41(11): 14-22.
[7]	范雁,杨淼,薛松. 基于光谱法-图像灰度法高通量筛选高效固定CO₂的苯甲酸脱羧酶^*[J]. 中国生物工程杂志, 2021, 41(11): 55-63.
[8]	梁爱玲,刘文婷,武攀,李倩,高健,张洁,刘卫东,贾士儒,郑迎迎. 来源于Exophiala aquamarina的新型玉米赤霉烯酮水解酶的性质及底物结合中心关键氨基酸的功能研究^*[J]. 中国生物工程杂志, 2021, 41(10): 19-27.
[9]	察亚平, 朱牧孜, 李爽. 体内连续定向进化研究进展 ^*[J]. 中国生物工程杂志, 2021, 41(1): 42-51.
[10]	郭二鹏, 张建志, 司同. 羊毛硫肽的高通量工程改造方法新进展 ^*[J]. 中国生物工程杂志, 2021, 41(1): 30-41.
[11]	徐晓, 程驰, 袁凯, 薛闯. 里氏木霉产纤维素酶研究进展 ^*[J]. 中国生物工程杂志, 2021, 41(1): 52-61.
[12]	蔺士新,刘东晨,雷云,熊盛,谢秋玲. TNF-α纳米抗体的筛选、表达及特异性检测 ^*[J]. 中国生物工程杂志, 2020, 40(7): 15-21.
[13]	朱衡,张继福,张云,胡云峰. 环氧交联剂和氨基载体固定化海洋假丝酵母脂肪酶^*[J]. 中国生物工程杂志, 2020, 40(5): 57-68.
[14]	马翠萍,刘朵朵,潘炳菊,申会涛,宋亚囝. 来源于嗜碱芽孢杆菌N16-5甘露聚糖利用基因簇的乙酰酯酶AesA的克隆及性质分析^*[J]. 中国生物工程杂志, 2020, 40(3): 65-71.
[15]	朱衡,张继福,张云,孙爱君,胡云峰. 聚乙二醇二缩水甘油醚交联氨基载体LX-1000EA固定化脂肪酶 ^*[J]. 中国生物工程杂志, 2020, 40(1-2): 124-132.

Viewed

Full text

Abstract

Cited

Shared

Discussed