-118-
2014 Vol.33 No.10 Serial No.272
China Brewing
酒曲中羧甲基纤维素酶活力测定影响因素分析
赵爽纪凤娣赵章林'宋吴'郑玉芝*艾金忠”胡军”（1.北京一轻研究院北京1011112.北京红星股份有限公司北京101400）
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摘要对还原糖法测定酒曲中羧甲基纤维素酶活力的影响因素进行了探讨建立了适合酒曲中羧甲基纤维素酶活的测定方法。研上清为原始酶液。酶解反应条件为pH值4.8样品为0.50mL酶液和2.00mL底物空白为0.50mL酶液和2.00mL缓冲液同时50℃反应 30min后添加3.0mLDNS试剂沸水浴显色10min冷却后空白补加2.00mL底物样品管补2.00mL缓冲液分别用水定客至25mL，测定波长540nm
关键词酒曲羧甲基纤维素醇活力提取酶解条件空白选择
中图分类号TS207.3
文献标识码A
doi :10.11882/jissn.02545071.2014.10.028
文章编号0254-5071(2014)10-0118-05
Analysis of affecting factors on determination of carboxymethyl cellulose activity in starter
ZHAO Shuang', JI Fengdi', ZHAO Zhanglin', SONG Hao', ZHENG Yuzhi*, AI Jinzhong', HU Jun
(1.Beijing Industria/ Technology Research Institute, Bejing 101 1 I I, China; 2.Bejing Red Star Co., Ltd., Bejing 101400)
Abstract: The influence factor of reducing sugar method on carboxyl methyl cellulose activity (CMCA) of starter was discussed and a testing method suitable for CMCA determination was established. The research focused on the effect of enzyme extraction method in starter (extraction ratio of buffer and starter, enzyme extraction methods of starter), enzymolysis reaction conditions(enzymatic reaction pH, reaction temperature, enzyme and substrate addition) and blank choice. Experimental results showed that the optimal detemination conditions were overnight cooling at -2o'C: seoooaoesaeoesoe solution. The reaction conditions of enzyme hydrolysis were as follows: 0.50 ml enzyme with 2.00 ml substrate as sample and 0.50 ml enzyme with 2.00 ml buffer solution as blank, reaction time 30 min, temperature 50°C and pH 4.8. Then 3.0 ml DNS reagent was added and water bathed for 10 min for chromogenic reaction. After cooling, the blank was supplemented with 2.00 ml substrate and the sample was supplemented with 2.00 ml buffes
solution, and then water were added to the solution to 25 ml. Finally the absorbance was determined at 540 nm. Key words: starter, carboxymethyl cellulose activity; extract; hydrolysis conditions; blank choice
固态白酒酒糟为白酒酿造过程中的主要副产物我国每年白酒丢糟产量都>2500万"。酒糟含有一定的蛋自质、脂肪以及粗纤维等说明固态自酒酒糟具有利用价值。依照循环经济及可持续发展思路固态白酒酒槽的资源化利用具有现实意义。据统计，白酒丢槽的粗纤维含量为 18%~24%其中包括纤维素和木质素。纤维素分子结构特殊难以被人类充分利用造成巨大的资源浪费和严重的环境污染如酒曲中的纤维素酶能将酒槽的纤维素水解为可发醇糖将提高酒槽的再利用率极有发展前景。
纤维素酶是一种多组分的复合酶近年来在食品工业、饲料工业生产及环保等领域中都有较大的应用价值)。秦广利等(3)应用纤维素酶对酱香型白酒资源化利用进行了研究在线槽中添加纤维素酶10U/g出酒率比原提高
收稿日期2014-07-09
了2.61%。岳思君等"筛选了有良好分解效果的纤维素分解菌。但是不同来源的纤维素酶组成及各组分比例有较大差异同时纤维素作用的底物也比较复杂致使纤维素酶活力的测定方法很多且方法复杂而不统一[5期。并且不同的测定条件对酶活力的最终测定结果有一定影响。目前常用的测定方法有羧甲基纤维素(还原糖法)酶活力测定方法、纸酶活力法和棉花糖化力法等。国内许多单位分别采用了以上客种方法并进行了种种修改使测定的方法更加多样化造成不同产品、不同结果之间无法相互比较的局面。目前尚未见测定酒曲中纤维素酶活力方法的研究。因酒曲实际是多种酶的复合制剂包括糖化酶、淀粉酶、纤维素酶等因此纯的酶制剂的测定方法不适用于酒曲中纤维素酶的测定羧甲基纤维素酶活力car-
作者简介赵爽(1988-）女士研究生研究方向为食品生物技术
*通讯作者郑玉芝（1965-）女教授级高级工程师博士研究方向为食品加工与检测
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