用于烟梗木质素降解的漆酶的分离纯化研究

张锐; 李元实; 寇霄腾; 朱良华; 余翔; 马林

doi:10.3969/j.issn.2095-476X.2014.06.002

CN 41-1437/TS ISSN 2096-1553

用于烟梗木质素降解的漆酶的分离纯化研究

张锐 , 李元实 , 寇霄腾 , 朱良华 , 余翔 , 马林

张锐, 李元实, 寇霄腾, 等. 用于烟梗木质素降解的漆酶的分离纯化研究[J]. 轻工学报, 2014, 29(6): 5-8,12. doi: 10.3969/j.issn.2095-476X.2014.06.002

引用本文: 张锐, 李元实, 寇霄腾, 等. 用于烟梗木质素降解的漆酶的分离纯化研究[J]. 轻工学报, 2014, 29(6): 5-8,12. doi: 10.3969/j.issn.2095-476X.2014.06.002

ZHANG Rui, LI Yuan-shi, KOU Xiao-teng, et al. Separation and purification of laccase used for tobacco stem lignin degradation[J]. Journal of Light Industry, 2014, 29(6): 5-8,12. doi: 10.3969/j.issn.2095-476X.2014.06.002

Citation: ZHANG Rui, LI Yuan-shi, KOU Xiao-teng, et al. Separation and purification of laccase used for tobacco stem lignin degradation[J]. Journal of Light Industry, 2014, 29(6): 5-8,12. doi: 10.3969/j.issn.2095-476X.2014.06.002

用于烟梗木质素降解的漆酶的分离纯化研究

张锐 ^1,4 ,
李元实 ² ,
寇霄腾 ³ ,
朱良华 ² ,
余翔 ¹ ,
马林 ^{1

,}

郑州轻工业学院烟草科学与工程学院, 河南郑州 450001;
吉林烟草工业有限责任公司生产技术中心, 吉林延吉 133000;
安徽中烟工业有限责任公司原料研究部, 安徽蚌埠 233000;
云南中烟再造烟叶有限责任公司技术中心, 云南昆明 650000

通讯作者: 马林

中图分类号: TS41⁺3

Separation and purification of laccase used for tobacco stem lignin degradation

College of Tobacco Science and Engineering, Zhengzhou University of Light Industry, Zhengzhou 450001, China;
Production and Technical Center, Jilin Tobacco Industry Co., Ltd., Yanji 133000, China;
Department of Materials Research, China Tobacco Anhui Industry Co., Ltd., Bengbu 233000, China;
Technology Center, China Tobacco Yunnan Reconstituted Tobacco Co., Ltd., Kunming 650000, China

Corresponding author: MA Lin,

Received Date: 2014-04-22
Available Online: 2014-11-15
CLC number: TS41⁺3

摘要: 通过云芝菌发酵得到的漆酶粗酶液经超滤浓缩后,以硫酸铵为盐析剂进行分级盐析沉淀,利用DEAE-Sepharose-F.F.柱层析对漆酶的发酵液进行分离纯化.SDS-PAGE电泳分析证明获得了电泳纯漆酶,其分子量约为64.4 kD.分离纯化过程中酶被纯化了22.80倍,酶活的回收率为36.32%,表明云芝菌产漆酶得到了很好的分离纯化.将漆酶液用于烟梗木质素的降解,实验结果表明,经此降解处理过的烟梗其内在品质得到了一定的提高:杂气和刺激性明显降低,木质气明显减弱,香气和余味均有改善.
- 烟梗木质素 /
- 漆酶 /
- 分离纯化 /
- 云芝菌
Abstract: After the laccase crude enzyme which was obtained by fermenting Coriolus versicolor had been concentrated and hyperfiltrated,grading salting precipitation and DEAE-Sepharose-F.F.column chromatography had been taken to separate and purify the laccase fermented liquid with ammonium sulfate as the salting-out agent.SDS-PAGE electrophoresis results showed that pure laccase could be obtained,the molecular weight was about 64.4 kD,enzyme was purified for 22.80 times by the purification process,the enzyme activity recovery rate was 36.32%.These proved that the laccase which was produced by Coriolus versicolor could be well purified.Using the enzyme solution in the experiment stem lignin degradation,the results showed that the intrinsic quality of the processed tobacco stems had been improved to some extent,miscellaneous gases and irritating significantly reduced,wood gas also significantly reduced,aroma and aftertaste slightly improved.
- tobacco stem lignin /
- laccase /
- separation and purification /
- Coriolus versicolor
1. [1]
  刘淑珍,钱世钧.担子菌漆酶的分离纯化及其性质研究[J].微生物学报,2003,43(1):73.
2. [2]
  Yosiiida H.Chemistry of lacquer (Urusht)[J].Chem Soc,1883,43:172.
3. [3]
  Bertrand G.Sur la presence simultanee de la laccase et de la tyrosinase dans le suc de quelques champignons[J].C R Hebd Seances Acad Sci,1987,123:463.
4. [4]
  Laborde J.Sur lacasse des vins[J].C R Hebd Seances Acad Sci,1896,123:1074.
5. [5]
  Reinhannar B.Laccase in Copper Proteins and Copper Enzymes[M].Boca Raton Ela:CRC Press,1984.
6. [6]
  汪家政,范明.蛋白质技术手册[M].北京:科学出版社,2000:243-260.
7. [7]
  Ryan S, Schnitzhofer W,Tzanov T,et al.An acid-stable laccase from Sclerotium rolfsii with potential for wool dye decolourization[J].Enzyme and Microbial Technology,2003, 33:766.
1. [1]
  贾尚羲 , 张怡雪 , 石盼盼 , 王昱 , 李可 . 不同时长超声波处理对鹰嘴豆分离蛋白乳化液稳定性的影响. 轻工学报, 2024, 39(5): 40-49. doi: 10.12187/2024.05.005
2. [2]
  张嫚 , 张国治 , 张康逸 , 何梦影 . 超声辅助酶解法制备小麦ACE抑制肽及其稳定性研究. 轻工学报, 2024, 0(0): -.
3. [3]
  张嫚 , 张国治 , 张康逸 , 何梦影 . 超声辅助酶解法制备小麦ACE抑制肽及其稳定性研究. 轻工学报, 2024, 39(5): 29-39. doi: 10.12187/2024.05.004
4. [4]
  卢晓波 , 徐海 , 朱俊召 , 张宇 , 谭健 , 高冠男 , 胡军华 , 林龙 . 基于机器视觉的加热卷烟烟支端部质量检测系统设计. 轻工学报, 2024, 0(0): -.

点击查看大图

计量

PDF下载量: 25
文章访问数: 821
引证文献数: 0

通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

张锐, 李元实, 寇霄腾, 等. 用于烟梗木质素降解的漆酶的分离纯化研究[J]. 轻工学报, 2014, 29(6): 5-8,12. doi: 10.3969/j.issn.2095-476X.2014.06.002

引用本文: 张锐, 李元实, 寇霄腾, 等. 用于烟梗木质素降解的漆酶的分离纯化研究[J]. 轻工学报, 2014, 29(6): 5-8,12. doi: 10.3969/j.issn.2095-476X.2014.06.002

用于烟梗木质素降解的漆酶的分离纯化研究

通讯作者: 马林

郑州轻工业学院烟草科学与工程学院, 河南郑州 450001;
吉林烟草工业有限责任公司生产技术中心, 吉林延吉 133000;
安徽中烟工业有限责任公司原料研究部, 安徽蚌埠 233000;
云南中烟再造烟叶有限责任公司技术中心, 云南昆明 650000

收稿日期: 2014-04-22
网络出版日期: 2014-11-15

关键词:

摘要: 通过云芝菌发酵得到的漆酶粗酶液经超滤浓缩后,以硫酸铵为盐析剂进行分级盐析沉淀,利用DEAE-Sepharose-F.F.柱层析对漆酶的发酵液进行分离纯化.SDS-PAGE电泳分析证明获得了电泳纯漆酶,其分子量约为64.4 kD.分离纯化过程中酶被纯化了22.80倍,酶活的回收率为36.32%,表明云芝菌产漆酶得到了很好的分离纯化.将漆酶液用于烟梗木质素的降解,实验结果表明,经此降解处理过的烟梗其内在品质得到了一定的提高:杂气和刺激性明显降低,木质气明显减弱,香气和余味均有改善.

English Abstract

Separation and purification of laccase used for tobacco stem lignin degradation

Corresponding author: MA Lin,

College of Tobacco Science and Engineering, Zhengzhou University of Light Industry, Zhengzhou 450001, China;
Production and Technical Center, Jilin Tobacco Industry Co., Ltd., Yanji 133000, China;
Department of Materials Research, China Tobacco Anhui Industry Co., Ltd., Bengbu 233000, China;
Technology Center, China Tobacco Yunnan Reconstituted Tobacco Co., Ltd., Kunming 650000, China

Received Date: 2014-04-22
Available Online: 2014-11-15

Abstract: After the laccase crude enzyme which was obtained by fermenting Coriolus versicolor had been concentrated and hyperfiltrated,grading salting precipitation and DEAE-Sepharose-F.F.column chromatography had been taken to separate and purify the laccase fermented liquid with ammonium sulfate as the salting-out agent.SDS-PAGE electrophoresis results showed that pure laccase could be obtained,the molecular weight was about 64.4 kD,enzyme was purified for 22.80 times by the purification process,the enzyme activity recovery rate was 36.32%.These proved that the laccase which was produced by Coriolus versicolor could be well purified.Using the enzyme solution in the experiment stem lignin degradation,the results showed that the intrinsic quality of the processed tobacco stems had been improved to some extent,miscellaneous gases and irritating significantly reduced,wood gas also significantly reduced,aroma and aftertaste slightly improved.