食品科学 ›› 2012, Vol. 33 ›› Issue (1): 125-130.doi: 10.7506/spkx1002-6630-201201025

• 生物工程 • 上一篇    下一篇

废弃食用菌栽培块中纤维素酶的提纯工艺研究

杜宁,许甜甜,李先富,夏志华,陈军   

  1. 上海师范大学生命与环境科学学院,上海 200234
  • 收稿日期:2011-02-15 修回日期:2011-12-26 出版日期:2012-01-15 发布日期:2012-01-12
  • 通讯作者: 陈军 E-mail:cj7206@163.com
  • 基金资助:

    国家自然科学基金项目(50678102;0000005472);上海市教委青年基金项目(03DQ25);
    上海师范大学科技基金项目(SK201228);上海市大学生科研创新活动计划项目(CX016)

Extraction and Purification of Cellulase from Spent Mushroom Compost

DU Ning,XU Tian-tian,LI Xian-fu,XIA Zhi-hua,CHEN Jun   

  1. College of Life and Environment Sciences, Shanghai Normal University, Shanghai 200234, China
  • Received:2011-02-15 Revised:2011-12-26 Online:2012-01-15 Published:2012-01-12
  • Contact: Chen Jun E-mail:cj7206@163.com

摘要:

利用废弃食用菌栽培块通过浸提、膜浓缩、盐析和双水相萃取等方法提取加工工业级和高纯度纤维素酶制剂。结果表明:采用含体积分数0.2% Triton X-100 的pH5.0、0.2mol/L 乙酸- 乙酸钠缓冲液作为酶浸提液,在25℃浸提3h 条件下可以抽提出102.64U/mL 的纤维素酶溶液,再在质量分数60% 的(NH4)2SO4 饱和度条件下可回收65.54% 的纤维素酶总活力,制备成酶活力达到2865.47U/mL 的工业级纤维素酶制剂;再经质量分数22% 聚乙二醇6000(PEG6000)-20%(NH4)2SO4 双水相体系在pH5.5、7mmol/L NaCl 条件下常温萃取,可获得酶活力为10208.46U/mL,酶比活力106.62U/mg 的高纯度纤维素酶,酶总活力得率为51.22%,纯度是浸提液的12.85 倍。

关键词: 废弃食用菌栽培块, 纤维素酶, 双水相萃取, 提取纯化

Abstract:

Spent mushroom compost (SMC) was used to prepare industrial grade cellulase by solvent extraction, membrane
filtration and salting out and high-purity cellulase by further aqueous two-phase extraction. The results showed that crude
cellulase solution with an enzymatic activity of 102.64 U/mL was obtained after 3 h of extraction at 25 ℃ with an extraction solvent
composed of 0.2 mol/L acetic-acetate sodium buffer at pH 5.0 and 0.2% Triton X-100. Subsequent salting out at 60% (NH4)2SO4
saturation could recover 65.54% of the total cellulase activity, and industrial grade cellulase with an enzymatic activity of
2865.47 U/mL was obtained. After further aqueous two-phase extraction with a system made up of 22% polyethylene glycol
6000 (PEG6000) and 20% (NH4)2SO4 under the conditions of 7 mmol/L NaCl, pH 5.5 and normal temperature, the cellulase activity
and specific activity were 10208.46 U/mL and 106.62 U/mg, respectively and a total activity yield of 51.22% was achieve,
revealing a 12.85-fold increase when compared with that of crude enzyme solution. As a result, high-purity cellulase was
obtained.

 

Key words: spent mushroom compost(SMC), cellulose, aqueous two-phase extraction, extraction and purification

中图分类号: