硬毛粗盖孔菌产漆酶条件优化及其对日落黄的降解作用
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摘要
以硬毛粗盖孔菌(Funalia trogii)为研究对象,考察培养基组成及培养条件等对其发酵产漆酶的影响,并进一步研究漆酶对食用合成色素日落黄的降解脱色效果。结果表明,最适培养基组成及培养条件为:玉米淀粉25 g/L,蛋白胨2 g/L,Cu~(2+)1.5 mmol/L,香兰素1 mmol/L,培养温度30℃、初始p H6.0,摇床转速200 r/min,硬毛粗盖孔菌培养8 d后发酵液漆酶活力为(329±12) U/m L,是初始培养条件下的11.96倍。漆酶粗酶液对日落黄呈现出较高的降解脱色作用并可耐受高浓度底物(800 mg/L),50～200 mg/L日落黄脱色率均可达90%以上,漆酶介体ABTS可提升对日落黄脱色效率及速率,并降低高浓度底物对漆酶的抑制作用,曲酸的加入则抑制了漆酶活性,导致漆酶对日落黄的脱色率仅为7.14%。本研究为漆酶资源在食品行业的深入开发与应用提供理论依据。
The effect of composition of medium and culture conditions on the laccase production by Funalia trogii was evaluated,and the degradation of edible synthetic pigment sunset yellow by crude laccase was further studied. The results showed that the optimum culture conditions were as follows: 25 g/L corn flour,2 g/L peptone,Cu~(2+) 1.5 mmol/L,1 mmol/L vanillin,culture temperature 30 ℃,initial pH6.0,rotation speed 200 r/min.Under the conditions of fermentation,the activity of laccase from F.trogii after cultured for eight days was( 329 ± 12) U/m L,which was 11.96-fold compared to the control.Crude laccase showed high efficiency degradation of sunset yellow and could tolerate high concentration of substrate( 800 mg/L).Decolorization efficiency for 50～200 mg/L sunset yellow more than 90%.The addition of laccase mediator could significantly increase the efficiency and rate of sunset yellow decolorization,meanwhile,decrease the inhibition of laccase by high concentration substrates.The addition of kojic acid inhibited the activity of laccase,then the decolorization rate of sunset yellow by laccase was only 7.14%.This study provides a theoretical basis for further development and application of laccase resources in the food industry.
The effect of composition of medium and culture conditions on the laccase production by Funalia trogii was evaluated,and the degradation of edible synthetic pigment sunset yellow by crude laccase was further studied. The results showed that the optimum culture conditions were as follows: 25 g/L corn flour,2 g/L peptone,Cu~(2+) 1.5 mmol/L,1 mmol/L vanillin,culture temperature 30 ℃,initial pH6.0,rotation speed 200 r/min.Under the conditions of fermentation,the activity of laccase from F.trogii after cultured for eight days was( 329 ± 12) U/m L,which was 11.96-fold compared to the control.Crude laccase showed high efficiency degradation of sunset yellow and could tolerate high concentration of substrate( 800 mg/L).Decolorization efficiency for 50～200 mg/L sunset yellow more than 90%.The addition of laccase mediator could significantly increase the efficiency and rate of sunset yellow decolorization,meanwhile,decrease the inhibition of laccase by high concentration substrates.The addition of kojic acid inhibited the activity of laccase,then the decolorization rate of sunset yellow by laccase was only 7.14%.This study provides a theoretical basis for further development and application of laccase resources in the food industry.
引文
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[19]吴香波,谢益民,冯晓静.白腐菌Coriolus versicolor的培养及产漆酶条件的研究[J].纤维素科学与技术,2009,17(2):12-19.
[20]李平,易弋,邓春,等.漆酶的结构及其应用研究进展[J].中国酿造,2016,36(5):10-15.
[21]柴新义,安双登,盛硕,等.产漆酶真菌筛选及其产酶条件的优化[J].西北农林科技大学学报:自然科学版,2015,43(3):205-210.
[22]廖海君,李蕊伽,陶敏,等.杂优-2平菇漆酶的分离纯化及酶学性质[J].食品科学,2016,37(19):147-153.
[23]李哲,艾柳英,李钊锋.金针菇生物降解与染料脱色作用研究[J].中国农学通报,2016,32(27):99-104.
[24]Emre B,Ozfer Y. Shirkabvand E. Enhanced production of laccase in repeated-batch cultures of Funalia trogii and Trametes versicolor[J].Biochemical Engineering Journal,2010,52:33-37.
[25]杨咏洁,吕蕾,王旻乾,等.火红密孔菌产漆酶培养基的优化[J].延边大学农学学报,2015,37(3):231-235.
[26]王寒,彭林,丁重阳,等.阿魏菇与胶红酵母共培养产漆酶对活性艳蓝W-RV的脱色[J].生物加工过程,2015,13(5):74-78.
[27]郑苗苗,焦战战,张令昂,等.红平菇产漆酶条件优化及对孔雀绿染料脱色的研究[J].微生物学杂志,2014,34(4):58-61.
[28]陶国翔,李爱秀,张敏,等.漆酶与ABTS间的结合模式与作用机制研究[J].化学与生物工程,2017,34(11):23-26.
[29]Afreen S,Bano F,Ahmad N,etal.Screening and optimization of laccase from cyanobacteria with its potential in ecolorization of anthraquinonic dye Remazol Brilliant Blue R[J].Biocatalysis and Agricultural Biotechnology.2017,10:403-410.
[2]Riva S. Laccases:Blue enzymes for green chemistry[J].TRENDS in Biotechnology,2006,24(5):219-226.
[3]袁海生,戴玉成,曹云,等.白腐真菌染料脱色菌株的筛选及一色齿毛菌脱色条件的研究[J].菌物学报,2010,29(3):429-436.
[4]张昱,赵倩,李冠华.漆酶在食品领域的应用研究进展[J].食品工业科技,2016,37(17):385-394.
[5]Giacobbe S,Pezzella C,Lettera V,et al.Laccase pretreatment for agrofood wastes valorization[J].Bioresource Technology,2018,265:59-65.
[6]安琪,司静,戴玉成.不同诱导培养基对糙皮侧耳液体发酵产漆酶活性的影响[J].菌物学报,2018,37(3):361-370.
[7]刘文华,蔡宇杰,范晶晶,等.毛栓菌产漆酶条件优化及该酶对合成染料脱色的特性[J].微生物学通报,2013,40(5):727-738.
[8]刘宇.食用合成色素日落黄和胭脂红的分析方法研究[D].西安:西北大学,2013.
[9]Fazeli S,Sohrabi B,Bagha A R.The study of Sunset Yellow anionic dye interaction with gemini and conventional cationic surfactants in aqueous solution[J]. Dyes and Pigments.2012,95:768-775.
[10]薛虎寅,尹永梅,张太昌,等.偶氮类合成色素检测技术的研究进展[J].生物技术进展,2012,2(3):171-176.
[11]戴玉成.中国储木及建筑木材腐朽菌图志[M].北京:科学出版社,2009:78-90.
[12]Li G H,Liu X X,Yuan L.Improved laccase production by Funalia trogii in absorbent fermentation with nutrient carrier[J].Journal of Bioscience and Bioengineering,2017,124(4):381-385.
[13]Wang S N,Chen Q J,Zhu M J.et al.An extracellular yellow laccase from white rot fungus Trametessp. F1635 and its mediator systems for dye decolorization[J].Biochimie,2018,148:46-54.
[14]孙瑞雪,杨萍,刘琦,等.虎皮香菇漆酶的发酵优化[J].食品研究与开发,2015,36(21):156-161.
[15]徐春燕,高迎荣,王晓瑜,等.胶质射脉革菌漆酶的复合诱导及其脱色性能[J].生物技术,2015,25(2):196-200.
[16]Kameshwar A,Barber R,Qin W S.Comparative modeling and molecular docking analysis of white,brown and soft rot fungal laccases using lignin model compounds for understanding the structural and functional properties of laccases[J]. Journal of Molecular Graphics and Modelling.2018,79:15-26.
[17]马瑛,贾红华,周华,等.高活性漆酶产生菌的发酵条件优化[J].南京工业大学学报:自然科学版,2012,32(2):82-86.
[18]陶君,马爱民,郭爱玲.灵芝漆酶活性的测定及其产漆酶条件的优化[J].食品科学,2008(3):310-313.
[19]吴香波,谢益民,冯晓静.白腐菌Coriolus versicolor的培养及产漆酶条件的研究[J].纤维素科学与技术,2009,17(2):12-19.
[20]李平,易弋,邓春,等.漆酶的结构及其应用研究进展[J].中国酿造,2016,36(5):10-15.
[21]柴新义,安双登,盛硕,等.产漆酶真菌筛选及其产酶条件的优化[J].西北农林科技大学学报:自然科学版,2015,43(3):205-210.
[22]廖海君,李蕊伽,陶敏,等.杂优-2平菇漆酶的分离纯化及酶学性质[J].食品科学,2016,37(19):147-153.
[23]李哲,艾柳英,李钊锋.金针菇生物降解与染料脱色作用研究[J].中国农学通报,2016,32(27):99-104.
[24]Emre B,Ozfer Y. Shirkabvand E. Enhanced production of laccase in repeated-batch cultures of Funalia trogii and Trametes versicolor[J].Biochemical Engineering Journal,2010,52:33-37.
[25]杨咏洁,吕蕾,王旻乾,等.火红密孔菌产漆酶培养基的优化[J].延边大学农学学报,2015,37(3):231-235.
[26]王寒,彭林,丁重阳,等.阿魏菇与胶红酵母共培养产漆酶对活性艳蓝W-RV的脱色[J].生物加工过程,2015,13(5):74-78.
[27]郑苗苗,焦战战,张令昂,等.红平菇产漆酶条件优化及对孔雀绿染料脱色的研究[J].微生物学杂志,2014,34(4):58-61.
[28]陶国翔,李爱秀,张敏,等.漆酶与ABTS间的结合模式与作用机制研究[J].化学与生物工程,2017,34(11):23-26.
[29]Afreen S,Bano F,Ahmad N,etal.Screening and optimization of laccase from cyanobacteria with its potential in ecolorization of anthraquinonic dye Remazol Brilliant Blue R[J].Biocatalysis and Agricultural Biotechnology.2017,10:403-410.