毛栓菌27-1漆酶对酚酸类化合物降解的效能分析

doi:10.11924/j.issn.1000-6850.casb2020-0770

中国农学通报 ›› 2021, Vol. 37 ›› Issue (13): 35-41.doi: 10.11924/j.issn.1000-6850.casb2020-0770

所属专题：生物技术；园艺

毛栓菌27-1漆酶对酚酸类化合物降解的效能分析

周鑫(), 缪晓磊, 余学军()

浙江农林大学/省部共建亚热带森林培育国家重点实验室,杭州 311300

收稿日期:2020-12-10 修回日期:2021-03-15 出版日期:2021-05-05 发布日期:2021-05-18
通讯作者: 余学军
作者简介:周鑫,男,1995年出生,浙江丽水人,硕士研究生,研究方向为竹林培育与利用。通信地址：311300 浙江省杭州市临安区武肃街666号浙江农林大学省部共建亚热带森林培育国家重点实验室,E-mail：305859941@qq.com。
基金资助:
浙江省省院合作重点项目“竹笋加工剩余物综合利用技术研究”(2017SY05)

The Degradation Efficiency of Trametes hirsuta 27-1 Laccase on Phenolic Acids

Zhou Xin(), Miao Xiaolei, Yu Xuejun()

State Key Laboratory of Subtropical Silviculture/ Zhejiang A&F University, Hangzhou 311300

Received:2020-12-10 Revised:2021-03-15 Online:2021-05-05 Published:2021-05-18
Contact: Yu Xuejun

摘要/Abstract

摘要：

探究毛栓菌27-1漆酶对酚酸类化合物的降解效能,为竹林地酚酸类自毒物质降解提供依据。采用超高效液相色谱检测法,通过测定菌液处理后酚酸含量变化,研究毛栓菌27-1漆酶对10种酚酸类化合物的降解特性。结果表明,毛栓菌27-1漆酶能完全降解酚酸类化合物中的香草酸、咖啡酸和芥子酸;丁香酸、阿魏酸和龙胆酸的降解效果一般,降解率50%左右;原儿茶酸的降解效果较差,降解率达35%;不能降解香豆酸、对羟基苯甲酸和绿原酸。研究不同条件下毛栓菌27-1漆酶降解酚酸类化合物丁香酸的效能,发现在pH 5、温度30℃、丁香酸初始浓度小于100 mg/L时,菌株对丁香酸具有高效降解能力,ABTS溶液作为介体物质加入降解体系后降解效果更好,在浓度为0.5 mmol/L时,降解率可达到100%。毛栓菌27-1产生的漆酶对酚酸类化合物具有良好降解效能。

关键词: 酚酸类化合物, 毛栓菌27-1, 漆酶, 竹林, 自毒物质, 降解效能, 丁香酸

Abstract:

The study aims to investigate the degradation efficiency of phenolic acid compounds by Trametes hirsuta 27-1 laccase so as to provide a basis for the degradation of phenolic acids in bamboo forest. Ultra-high performance liquid chromatography was used. By measuring the change of phenolic acid content after bacterial solution treatment, we explored the degradation properties of 10 kinds of phenolic acid compounds by Trametes hirsuta 27-1 laccase. The results show that laccase could completely degrade vanillic acid, caffeic acid and sinapic acid; the degradation effect of syringic acid, ferulic acid and gentian acid are general and the degradation rate is about 50%; the degradation effect of protocatechuic acid is poor, the degradation rate could be 35%, and coumaric acid, p-hydroxybenzoic acid and chlorogenic acid could not be degraded. By studying the efficacy of Trametes hirsuta 27-1 laccase in the degradation of syringic acid under different conditions, it is found that, under the condition of pH 5, temperature of 30℃ and the initial concentration of syringic acid less than 100 mg/L, the strain could have high degradation ability of syringic acid. After ABTS solution is added to the degradation system as a mediator, the degradation effect is much better. At a concentration of 0.5 mmol/L, the degradation rate could reach 100%. In conclusion, the laccase produced by Trametes hirsuta 27-1 has good degradation efficiency on phenolic acids.

Key words: phenolic acid, Trametes hirsuta 27-1, laccase, bamboo forest, autotoxins, degradation efficiency, syringic acid

中图分类号:

周鑫, 缪晓磊, 余学军. 毛栓菌27-1漆酶对酚酸类化合物降解的效能分析[J]. 中国农学通报, 2021, 37(13): 35-41.

Zhou Xin, Miao Xiaolei, Yu Xuejun. The Degradation Efficiency of Trametes hirsuta 27-1 Laccase on Phenolic Acids[J]. Chinese Agricultural Science Bulletin, 2021, 37(13): 35-41.

图/表 8

图1. 10种酚酸标准样品和供式菌液处理样品色谱图 1.原儿茶酸,2.龙胆酸,3.对羟基苯甲酸,4.绿原酸,5.香草酸,6.咖啡酸,7.丁香酸,8.对香豆酸,9.阿魏酸,10.芥子酸

化合物	标准曲线	R²	线性范围/(μg/L)
原儿茶酸	Y=4.2654X-1.9390	0.99985	0.064~1.280
龙胆酸	Y=2.1431X-2.3371	0.99984	0.021~0.820
对羟基苯甲酸	Y=3.9147X-3.6120	0.99985	0.032~0.640
绿原酸	Y=2.2957X-2.3821	0.99983	0.026~0.520
香草酸	Y=8.9511X-4.6955	0.99987	0.028~1.560
咖啡酸	Y=10.5631X-12.4554	0.99981	0.065~2.598
丁香酸	Y=2.4513X-2.6354	0.99984	0.015~0.542
对香豆酸	Y=6.3682X-4.7442	0.99985	0.047~1.231
阿魏酸	Y=2.9674X-4.3961	0.99986	0.021~1.328
芥子酸	Y=8.6588X-4.5453	0.99986	0.043~0.965

表1. 化合物的标准曲线、相关系数、线性范围

图2. 毛栓菌27-1漆酶对不同酚酸底物的降解效能

液体	0 h	4 h	8 h	12 h	24 h
灭活酶液	100.00	100.00	99.84	99.62	98.41
粗酶液	100.00	98.44	54.79	47.41	47.32
等体积超纯水	100.00	100.00	100.00	100.00	100.00

表2. 不同液体对丁香酸降解效能（残留率） %

图3. 不同温度对毛栓菌27-1漆酶降解效能的影响

图4. 不同初始pH对毛栓菌27-1漆酶降解效能的影响

图5. 不同初始浓度丁香酸对毛栓菌27-1漆酶降解效能的影响

图6. 不同浓度介体ABTS溶液对毛栓菌27-1降解效能的影响图中不同小写字母表示差异显著性(P<0.05)

参考文献 26

[1]	高贵宾, 潘雁红, 吴良如, 等. 不同覆盖栽培年限雷竹林生物量分配格局研究[J]. 江西农业大学学报, 2015,37(4):663-669.
[2]	王海霞, 曾庆南, 程平. 不同覆盖材料及厚度对雷竹笋生产的影响研究[J]. 南方林业科学, 2020,48(4):34-39.
[3]	杨开, 杨振寰, 吴伟杰, 等. 雷笋膳食纤维酶法改性及其理化性能和结构变化[J]. 食品与发酵工业, 2019,45(4):36-41.
[4]	袁娜. 覆盖雷竹林土壤生化性质及其变化动态[D]. 北京:中国林业科学研究院, 2009.
[5]	Dashtban M, Schraft H, Syed, et al. Fungal biodegradation and enzymatic modification of lignin[J]. International Journal of Biochemistry and Molecular Biology, 2010,1(1):36-50. pmid: 21968746
[6]	Pooja U, Rahul S, Pavan K A. Bioprospecting and biotechnological applications of fungal laccase[J]. Biotech, 2016,6(1):15.
[7]	Peng X, Yuan X Z, Liu H, et al. Degradation of Polycyclic Aromatic Hydrocarbons (PAHs) by Laccase in Rhamnolipid Reversed Micellar System[J]. Applied Biochemistry and Biotechnology, 2015,176(1). 45-55. doi: 10.1007/s12010-015-1508-3 pmid: 25637508
[8]	罗爽, 谢天, 刘忠川, 等. 漆酶/介体系统研究进展[J]. 应用与环境生物学报, 2015,21(6):987-995.
[9]	沈琦, 骆镜, 陈旖凡, 等. 生物炭固定杏鲍菇漆酶降解废水中的苯胺探究[J]. 南方农业, 2019,13(30):125-126.
[10]	谭叶林, 杜全能, 朱文娟, 等. 产漆酶细菌筛选鉴定及固体发酵条件研究[J]. 安徽农业科学, 2020,48(1):1-6.
[11]	Zhang J, Ren X, Chen W Q, et al. Biological pretreatment of corn stover by solid state fermentation of Phanerochaete chrysosporium[J]. Frontiers of Chemical Science and Engineering, 2012,6(2):146-151. doi: 10.1007/s11705-012-1220-6 URL
[12]	Hilgers R, Kabel M A, Vincken J P. On the reactivity of p-coumaroyl groups in lignin upon laccase and laccase/HBT treatments[J]. ACS Sustainable Chemistry & Engineering, 2020,8(23):8723-8731.
[13]	Ahmet U, Emre Bi, Filiz B, et al. Laccase-conjugated thiolated chitosan-Fe₃O₄ hybrid composite for biocatalytic degradation of organic dyes[J]. International Journal of Biological Macromolecules, 2020,150(1):871-884. doi: 10.1016/j.ijbiomac.2020.02.006 URL
[14]	朱虹, 王宏伟, 杜威, 等. 内生真菌重组漆酶rLACB3修复花生连作土壤[J]. 生态学杂志, 2014,33(7):1920-1927.
[15]	李新鑫, 余学军, 俞暾, 等. 竹林环境中木质素降解菌株的分离鉴定[J]. 竹子学报, 2016,35(2):20-25.
[16]	Arora D S, Gill P K . Effects of various media and supplements on laccase production by some white rot fungi[J]. Bioresource Technology, 2001,77(1):8-91.
[17]	陈芸, 马刘峰. 丁香酸和阿魏酸对大豆种子萌发及幼苗生长的影响[J]. 种子, 2017,36(1):108-111.
[18]	刘忠川, 王刚刚. 真菌漆酶结构与功能研究进展[J]. 生物物理学报, 2013,29(9):629-645.
[19]	林鹏飞, 贾小舟, 祁燕, 等. 酚酸类化合物研究进展[J]. 广东化工, 2017,44(1):50-52.
[20]	朱洪玲, 傅佳佳, 许波, 等. 漆酶催化4-氨基苯酚聚合[J]. 精细化工, 2018,35(4):620-624.
[21]	Eresa A, Jose G, Pazos M, et al. Enhanced production of laccase in Coriolopsis rigida grown on barley bran in flask or expanded-bed bioreactor[J]. World Journal of Microbiology & Biotechnology, 2007,23(8):1189-1194. doi: 10.1007/s11274-006-9334-y URL
[22]	Dedeyan B, Klonowsk A, Tagger S, et al. Biochemical and Molecular Characterization of a Laccase from Marasmius quercophilus[J]. Applied and Environmental Microbiology, 2000,66(3):925-929. doi: 10.1128/AEM.66.3.925-929.2000 URL
[23]	Gu Y H, Xue P, Jia F, et al. Co-immobilization of laccase and ABTS onto novel dual-functionalized cellulose beads for highly improved biodegradation of indole[J]. Journal of Hazardous Materials, 2019,365(1). 118-124 doi: 10.1016/j.jhazmat.2018.10.076 URL
[24]	Yanto D H Y, Auliana N, Anita S H, et al. Decolorization of synthetic textile dyes by laccase from newly isolated Trametes hirsuta EDN084 mediated by violuric acid[J]. IOP Conference Series: Earth and Environmental ence, 2019,374(1):012005.
[25]	Valls C, José F. C, Baffert C, et al. Comparing the efficiency of the laccase-NHA and laccase-HBT systems in eucalyptus pulp bleaching[J]. Biochemical Engineering Journal, 2010,49(3):401-407. doi: 10.1016/j.bej.2010.02.002 URL
[26]	徐鑫, 张国庆, 胡渤洋, 等. 真菌漆酶及其介体系统:来源、机理与应用[J]. 生物技术进展, 2020,10(1):30-39.

毛栓菌27-1漆酶对酚酸类化合物降解的效能分析

The Degradation Efficiency of Trametes hirsuta 27-1 Laccase on Phenolic Acids

RichHTML

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

图/表 8

参考文献 26

相关文章 11

编辑推荐

Metrics

本文评价

关于本刊

作者中心

审者中心

在线期刊

联系我们

[1]	刘海英, 王增, 徐耀文, 葛晓改, 周本智, 蒋仲龙. 不同林分密度毛竹林生态系统碳储量特征[J]. 中国农学通报, 2022, 38(35): 17-21.
[2]	刘晓婷,郭九峰. 低温对蒙古口蘑菌丝体漆酶活性及其基因家族表达的影响[J]. 中国农学通报, 2019, 35(3): 40-46.
[3]	耿贵,杨瑞瑞,吕春华,李任任,王宇光. 作物连作障碍研究进展[J]. 中国农学通报, 2019, 35(10): 36-42.
[4]	金晓婷,张志勇,杨栋,余向阳. 阿魏侧耳(Pleuratus ferulae)产漆酶条件的优化及其酶学性质研究[J]. 中国农学通报, 2016, 32(33): 151-156.
[5]	杨雪梅刘彦中刘志莲赖泳红陈勇能. 产漆酶菌株的筛选及其对烟秆降解效果的研究[J]. 中国农学通报, 2014, 30(7): 52-57.
[6]	何志刚汪仁王秀娟牛世伟李波. 花生自毒物质降解菌的筛选及其降解效果初步研究[J]. 中国农学通报, 2014, 30(21): 224-227.
[7]	谢国雄沈建国王京文张奇春侯昌萍梁颖颖. 不同经营措施下毛竹林土壤有机碳组分研究[J]. 中国农学通报, 2013, 29(31): 53-57.
[8]	官凤英范少辉郜燕芳肖复明蔡华利. 不同分类方法在竹林遥感信息识别中的应用[J]. 中国农学通报, 2013, 29(1): 47-52.
[9]	桂玲孙华陈利. 基于中等分辨率遥感影像的桃源县竹林信息提取研究[J]. 中国农学通报, 2012, 28(1): 85-91.
[10]	高玉千,高方,张世敏,吴坤. 黑木耳漆酶高产菌株的筛选[J]. 中国农学通报, 2009, 25(21): 301-304.
[11]	万善霞,滑静,王文平,张淑萍. 杏鲍菇漆酶部分酶学性质的研究[J]. 中国农学通报, 2009, 25(21): 107-109.