Please wait a minute...
 
 
食品与发酵工业  2021, Vol. 47 Issue (9): 70-75    DOI: 10.13995/j.cnki.11-1802/ts.025435
  研究报告 本期目录 | 过刊浏览 | 高级检索 |
中性脲酶固定化降解黄酒中尿素
李童1,2, 钱斌3, 周建弟3, 徐岩1,2, 王栋1,2*
1(江南大学 生物工程学院,江苏 无锡,214122)
2(教育部工业生物技术重点实验室(江南大学),江苏 无锡,214122)
3(古越龙山绍兴酒股份有限公司技术中心,浙江 绍兴,312000)
Degradation of urea in Huangjiu by immobilized neutral urease
LI Tong1,2, QIAN Bin3, ZHOU Jiandi3, XU Yan1,2, WANG Dong1,2*
1(School of Biotechnology,Jiangnan University,Wuxi 214122,China)
2(The Key Laboratory of Industrial Biotechnology,Ministry of Education,Jiangnan University,Wuxi 214122,China)
3(Gu Yue Longshan Shaoxing Wine Co.,Ltd.Technology Center,Shaoxing 312000,China)
下载:  HTML  PDF (1886KB) 
输出:  BibTeX | EndNote (RIS)      
摘要 为了探究固定化中性脲酶用于黄酒中尿素降解的可行性,首先对游离中性脲酶的酶学性质进行了测定,并与酸性脲酶进行比较,分析其用于处理黄酒中尿素的潜力;然后以壳聚糖为载体采用共价结合法进行中性脲酶的固定化,将固定化中性脲酶用于黄酒中尿素的降解。结果表明,游离中性脲酶在酸性条件(pH 4.0)下只能保留极低的酶活力,较高的酒精体积分数(24%)对其酶活力的影响不大。经过固定化,中性脲酶的最适pH由7.0降低到5.0,耐酸性也有显著提高。与游离酶相比,温度和酒精对于固定化酶活性和稳定性的影响没有显著改变。将此固定化中性脲酶处理黄酒,在30 ℃,加酶量为1 000 U/L的条件下处理24 h,可减除黄酒中95%以上的尿素。重复利用固定化酶处理黄酒也有一定的效果。该研究结果为今后固定化中性脲酶用于黄酒中尿素的降解提供了一定依据。
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
李童
钱斌
周建弟
徐岩
王栋
关键词:  黄酒  尿素  中性脲酶  固定化酶    
Abstract: In order to explore the feasibility of immobilized neutral urease for the degradation of urea in Huangjiu (Chinese rice wine), the enzymatic properties of free neutral urease were measured and compared with that of the acid urease. The covalent bonding method was used to immobilize neutral urease with chitosan, and the immobilized neutral urease was used for the degradation of urea in Huangjiu. The results showed that the free neutral urease retained very low enzyme activity under acidic conditions (pH=4.0), while high ethanol volume fraction (24%) had little effect on its activity. After immobilization, the optimal pH of neutral urease was lowered down from 7.0 to 5.0, and acid resistance was also significantly improved. Compared with the free enzyme, temperature and alcohol had little effect on the activity and stability of the immobilized enzyme. This immobilized neutral urease was used to treat Huangjiu at 30 ℃ with dosage of 1 000 U/L for 24 hours, which could degrade more than 95% of urea in Huangjiu. Reusing immobilized enzymes also had a certain effect on the treatment of Huangjiu. This study provides a basis for the future application of immobilized neutral urease for the degradation of urea in Huangjiu.
Key words:  Huangjiu(Chinese rice wine)    urea    neutral urease    immobilized urease
收稿日期:  2020-08-21      修回日期:  2020-09-18           出版日期:  2021-05-15      发布日期:  2021-06-03      期的出版日期:  2021-05-15
基金资助: 国家重点研发计划项目(2018YFC1604106);国家轻工技术与工程一流学科自主课题(LITE2 018-12);江苏高校品牌专业建设工程资助项目(TAPP)
作者简介:  硕士研究生(王栋教授为通讯作者,E-mail:dwang@jiangnan.edu.cn)
引用本文:    
李童,钱斌,周建弟,等. 中性脲酶固定化降解黄酒中尿素[J]. 食品与发酵工业, 2021, 47(9): 70-75.
LI Tong,QIAN Bin,ZHOU Jiandi,et al. Degradation of urea in Huangjiu by immobilized neutral urease[J]. Food and Fermentation Industries, 2021, 47(9): 70-75.
链接本文:  
http://sf1970.cnif.cn/CN/10.13995/j.cnki.11-1802/ts.025435  或          http://sf1970.cnif.cn/CN/Y2021/V47/I9/70
[1] FORKER T P G.Mechanisms of lung tumorigenesis by ethyl carbamate and vinyl carbamate[J].Drug Metabolism Reviews,2010,42(2):355-378.
[2] WEBER J V,SHARYPOV V I.Ethyl carbamate in foods and beverages:A review[J].Environmental Chemistry Letters,2009,7(3):233-247.
[3] ZHAO X R,DU G C,ZOU H J,et al.Progress in preventing the accumulation of ethyl carbamate in alcoholic beverages[J].Trends in Food Science&Technology,2013,32(2):97-107.
[4] GOWD V,SU H M,KARLOVSKY P,et al.Ethyl carbamate:An emerging food and environmental toxicant[J].Food Chemistry,2018,248:312-321.
[5] LACHENMEIER D W.Consequences of IARC re-evaluation of alcoholic beverage consumption and ethyl carbamate on food control[J].Deutsche Lebensmittel-Rundschau,2007,103(7):307-311.
[6] JIAO Z H,DONG Y C,CHEN Q H.Ethyl carbamate in fermented beverages:presence,analytical chemistry,formation mechanism,and mitigation proposals[J].Comprehensive Reviews in Food Science and Food Safety,2014,13(4):611-626.
[7] FAN J,LEI J,WANG L,et al.Rapid and high-capacity immobilization of enzymes based on mesoporous silicas with controlled morphologies[J].Chemical Communications,2003,(17):2 140-2 141.
[8] LIU J,XU Y,NIE Y,et al.Optimization production of acid urease by Enterobacter sp.in an approach to reduce urea in Chinese rice wine[J].Bioprocess and Biosystems Engineering,2012,35(4):651-657.
[9] YANG Y Q,KANG Z,ZHOU J L,et al.High-level expression and characterization of recombinant acid urease for enzymatic degradation of urea in rice wine[J].Applied Microbiology and Biotechnology,2015,99(1):301-308.
[10] ANDRICH L,ESTI M,MORESI M.Urea degradation in model wine solutions by free or immobilized acid urease in a stirred bioreactor[J].Journal of Agricultural and Food Chemistry,2009,57(9):3 533-3 542.
[11] ANDRICH L,ESTI M,MORESI M.Urea degradation kinetics in model wine solutions by acid urease immobilised onto chitosan-derivative beads of different sizes[J].Enzyme and Microbial Technology,2010,46(5):397-405.
[12] 查小红,杨广明,田亚平.一种天然材料复合体系固定化酒用双功能酶的研究[J].食品工业科技,2014,35(14):186-191;196.
ZHA X H,YANG G M,TIAN Y P.Study on an immobilized bifunctional enzyme applied in rice wine with natural materials[J].Science and Technology of Food Industry,2014,35(14):186-191;196.
[13] YANG L,LIU X,ZHOU N,et al.Characteristics of refold acid urease immobilized covalently by graphene oxide-chitosan composite beads[J].Journal of Bioscience and Bioengineering,2019,127(1):16-22.
[14] 范文来,王栋.近10年我国传统饮料酒白酒和黄酒品质安全研究现状与展望[J].食品安全质量检测学报,2019,10(15):4 811-4 829.
FAN W L,WANG D.Current practice and future trends of alcoholic beverages safety of China traditional Baijiu and Huangjiu in recent decades[J].Journal of Food Safety & Quality,2019,10(15):4 811-4 829.
[15] LIU Q T,CHEN Y Q,YUAN M L,et al.A Bacillus paralicheniformis iron-containing urease reduces urea concentrations in rice wine[J].Applied and Environmental Microbiology,2017,83(17).DOI:10.1128/aem.01258-17.
[16] MONIER M,EL-SOKKARY A M A.Modification and characterization of cellulosic cotton fibers for efficient immobilization of urease[J].International Journal of Biological Macromolecules,2012,51(1-2):18-24.
[17] LV M M,MA X F,ANDERSON D P,et al.Immobilization of urease onto cellulose spheres for the selective removal of urea[J].Cellulose,2018,25(1):233-243.
[18] ALATAWI F S,MONIER M,ELSAYED N H.Amino functionalization of carboxymethyl cellulose for efficient immobilization of urease[J].International Journal of Biological Macromolecules,2018,114:1 018-1 025.
[19] 高雅.黄酒中氨基甲酸乙酯降低措施研究[D].合肥:合肥工业大学,2017.
GAO Y.Study on reduction measures of ethyl carbamate in Chinese rice wine[D].Hefei:Hefei University of Technology,2017.
[20] KUMAR S,DWEVEDI A,KAYASTHA A M.Immobilization of soybean (Glycine max) urease on alginate and chitosan beads showing improved stability:Analytical applications[J].Journal of Molecular Catalysis B-Enzymatic,2009,58(1-4):138-145.
[21] WEATHERBURN M W.Phenol-hypochlorite reaction for determination of ammonia[J].Analytical Chemistry,1967,39(8):971-974.
[22] CLARK S,FRANCIS P S,CONLAN X A,et al.Determination of urea using high-performance liquid chromatography with fluorescence detection after automated derivatisation with xanthydrol[J].Journal of Chromatography A,2007,1 161(2):207-213.
[23] 郭双丽.中国黄酒贮存过程中尿素对氨基甲酸乙酯形成的影响及其物理减除技术研究[D].无锡:江南大学,2016.
GUO S L.The effect of urea on ethyl carbamate formation during rice wine storage and its removal by resin adsorption[D].Wuxi:Jiangnan University,2016.
[24] 国家市场监督管理总局,中国国家标准化管理委员会.GB/T 13662—2018 黄酒[S].北京:中国标准出版社,2018.
State Administration for Market Regulation,Standardization Administration.GB/T 1366—2018 Huangjiu[S].Beijing:China Standards Press China Standards Press,2018.
[25] 赵凯,许鹏举,谷广烨.3,5-二硝基水杨酸比色法测定还原糖含量的研究[J].食品科学,2008,29(8):534-536.
ZHAO K,XU P J,GU G Y.Study on determination of reducing sugar content using 3,5-dinitrosalicylic acid method[J].Food Science,2008,29(8):534-536.
[26] NARENDRANATH N V,THOMAS K C,INGLEDEW W M.Effects of acetic acid and lactic acid on the growth of Saccharomyces cerevisiae in a minimal medium[J].Journal of Industrial Microbiology and Biotechnology,2001,26(3):171-177.
[1] 刘梦, 缪礼鸿, 刘蒲临, 王霜, 高瑞杰. 马克斯克鲁维酵母与酿酒酵母混合发酵对液态法黄酒风味的影响[J]. 食品与发酵工业, 2021, 47(9): 160-167.
[2] 史瑛, 冯欣静, 周志磊, 姬中伟, 徐岳正, 毛健. 黄酒多糖对炎症性肠病及便秘作用机制的研究进展[J]. 食品与发酵工业, 2021, 47(9): 275-283.
[3] 张波, 谢广发, 李国龙, 孙国昌, 金建明, 朱炜俊, 刘菊. 黄酒生物酸化浸米与浸米浆水的利用[J]. 食品与发酵工业, 2021, 47(7): 168-174.
[4] 蒋彰, 周志磊, 姬中伟, 韩吉臣, 毛健, 周哲敏. 即墨黄酒煮糜工艺对挥发性物质的影响[J]. 食品与发酵工业, 2021, 47(5): 86-91.
[5] 宁亚维, 侯琳琳, 于同月, 刘茁, 杨正, 王志新, 贾英民. UPLC-MS/MS法快速测定乳酸菌发酵食品中的苯乳酸[J]. 食品与发酵工业, 2021, 47(5): 174-179.
[6] 宗原, 刘登峰, 刘以安. 基于改进蚁狮优化算法的黄酒发酵过程模型的参数辨识[J]. 食品与发酵工业, 2021, 47(2): 153-159.
[7] 马瑞娟, 林煌华, 谢友坪, 陈剑锋. 固定化酶制备鳀鱼蒸煮液蛋白肽及其性能表征[J]. 食品与发酵工业, 2020, 46(9): 122-127.
[8] 胡武瑶, 杨昳津, 窦慧, 阿依妮尕尔·约麦尔, 陈俊达, 王昭灵, 艾连中, 俞剑燊, 夏永军. 不同麦曲酿造黄酒中挥发性风味物质的代谢差异[J]. 食品与发酵工业, 2020, 46(8): 226-233.
[9] 白静, 王会. 红外光谱法测定奶粉中三聚氰胺和尿素[J]. 食品与发酵工业, 2020, 46(8): 267-272.
[10] 吴殿辉, 李晓敏, 蔡国林, 孙军勇, 谢广发, 陆健. 低产尿素黄酒酵母工程菌的酿造特性[J]. 食品与发酵工业, 2020, 46(3): 1-7.
[11] 郑瑞龙, 索婧怡, 钮成拓, 郑飞云, 易崇华, 圣弟青, 王金晶, 李崎, 刘春凤. 紫(黑)米在酿酒行业中的应用[J]. 食品与发酵工业, 2020, 46(23): 263-268.
[12] 刘蓉, 栾春光, 王德良, 覃思, 郝飞克. 基于高通量测序分析黄酒对D-半乳糖致衰老小鼠模型肠道微生物菌群的影响[J]. 食品与发酵工业, 2020, 46(2): 32-39.
[13] 孔令华, 夏小乐, 辛瑜. 甜型黄酒陈酿过程中5-羟甲基糠醛的生成规律[J]. 食品与发酵工业, 2020, 46(12): 258-263.
[14] 谢广发, 蔡际豪, 钱斌, 王兰, 鲁振东, 吕飞, 丁玉庭. 机械化大罐发酵干型黄酒酿造过程中风味物质的变化[J]. 食品与发酵工业, 2020, 46(11): 157-164.
[15] 彭立影, 刘功良, 许莹莹, 余洁瑜, 白卫东, 贾爱娟, 何松贵. 蜂蜜接合酵母协同传统曲种发酵广东黄酒的工艺研究[J]. 食品与发酵工业, 2019, 45(8): 103-108.
No Suggested Reading articles found!
Viewed
Full text


Abstract

Cited

  Shared   
  Discussed   
版权所有 © 《食品与发酵工业》编辑部
地址:北京朝阳区酒仙桥中路24号院6号楼111室
本系统由北京玛格泰克科技发展有限公司设计开发  技术支持:support@magtech.com.cn