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。
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.
出处
《食品科学》
EI
CAS
CSCD
北大核心
2012年第1期125-130,共6页
Food Science
基金
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)
关键词
废弃食用菌栽培块
纤维素酶
双水相萃取
提取纯化
spent mushroom compost（SMC）
cellulose
aqueous two-phase extraction
extraction and purification
