诱导剂对乳白耙齿菌产MnP活性影响研究
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摘要
为显著提高白腐菌(Irpex lacteus)产锰过氧化物酶(MnP)的活性。在液体培养条件下用分光光度计法检测12种诱导剂对乳白耙齿菌MnP活性的诱导效果。其中化学诱导剂包括愈创木酚、苯酚、香兰素、藜芦醇、苯甲酸、没食子酸、酪氨酸、二甲苯、苯胺蓝和ABTS,生物诱导剂包括麦麸和木屑。乳白耙齿菌在28℃下分别加入12种诱导剂进行培养,得到了不同时间的培养液,然后检测培养液中Mn P活性,从而得到乳白耙齿菌产MnP活性与诱导剂之间的关系。在初始培养基中加入0.2 mmol/L Mn~(2+)的培养条件下,诱导剂麦麸和ABTS对Mn P的诱导效果极为明显,二者在添加量分别为1.5%和0.2 mmol/L时,MnP活力分别为104.2 U/L和145.2 U/L,是初始产酶培养基酶活的8.6倍和11倍;而两种诱导剂组合,即同时添加1.5%麦麸与0.2 mmol/L ABTS,MnP活力为203.2 U/L,是初始培养条件MnP活性的21.9倍,实验表明诱导剂麦麸和ABTS能大幅度地提高白腐菌产MnP的活性。
In order to significantly increase the activity of Mn-P-producing bacteria( Mn P) in white rot fungi Irpex lacteus,12 kinds of inducers were detected by spectrophotometry under liquid culture conditions.The chemical inducers were guaiacol,phenol,vanillin,resveratrol,benzoic acid,gallic acid,tyrosine,xylene,aniline blue,ABTS,and the bio-inducer were wheat bran,wood chips.The activity of Mn P in the culture medium was measured by adding 12 kinds of inducers at 28 ℃,and the relationship between the activity of MnP and the inducer was obtained.0.2 mmol/L Mn~(2+) was added to the initial medium,and the effect of wheat bran and ABTS on MnP were obvious.The optimum additions were 1.5% and 0.2 mmol/L,respectively.Under these conditions the activity of MnP were 8.6 and 11 times that of the initial enzyme-producing medium.The two inducers were added with 1.5% of wheat bran and 0.2 mmol/L ABTS,the activity of the enzyme was 203.2 U/L,which was 21.9 times that under the initial culture condition.The results showed that the inducible wheat bran and ABTS could significantly increase the MnP activity of Irpex lacteus.
In order to significantly increase the activity of Mn-P-producing bacteria( Mn P) in white rot fungi Irpex lacteus,12 kinds of inducers were detected by spectrophotometry under liquid culture conditions.The chemical inducers were guaiacol,phenol,vanillin,resveratrol,benzoic acid,gallic acid,tyrosine,xylene,aniline blue,ABTS,and the bio-inducer were wheat bran,wood chips.The activity of Mn P in the culture medium was measured by adding 12 kinds of inducers at 28 ℃,and the relationship between the activity of MnP and the inducer was obtained.0.2 mmol/L Mn~(2+) was added to the initial medium,and the effect of wheat bran and ABTS on MnP were obvious.The optimum additions were 1.5% and 0.2 mmol/L,respectively.Under these conditions the activity of MnP were 8.6 and 11 times that of the initial enzyme-producing medium.The two inducers were added with 1.5% of wheat bran and 0.2 mmol/L ABTS,the activity of the enzyme was 203.2 U/L,which was 21.9 times that under the initial culture condition.The results showed that the inducible wheat bran and ABTS could significantly increase the MnP activity of Irpex lacteus.
引文
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[13]李鑫,李建科,董攀,等.白腐菌发酵培养及诱导剂对漆酶合成的影响[J].食品工业科技,2013,3(5):160-168.Li X,Li J K,Dong P,et al.White-rot fungus fermentation cultivation and effect of inducer on laccase production[J]. Science and Technology of Food Industry,2013,3(5):160-168.
[14]Feng X Y,Chen H Y,Xue D S,et al.Enhancement of Laccase activity by marine-derived Deuteromycete pestalotiopsis sp. J63with agricultural residues and inducers[J]. Biotechnology and Bioengineering,Chinese Journal of Chemical Engineering,2013,21(10):1182-1189.
[15]王忠燕,于萍,陈艳秋.白耙齿菌最佳培养基配方[J].食用菌学报,2013,20(3):15-17.Wang Z Y,Yu P,Chen Y Q.Optimization of selected nutritional parameters affecting the vegetative growth of Irpex lacteus[J].Acta Edulis Fungi,2013,20(3):15-17.
[16]杨晓宽,杜连祥,路福平.白腐菌产锰过氧化物酶培养基的优化[J].生物技术,2004,14(3):49-50.Yang X K,Du L X,Lu F P.Optimization of medium for manganese peroxidase production by white rot fungi[J].Biotechnology,2004,14(3):49-50.
[2]李玉,图力古尔.中国长白山蘑菇[M].北京:科学出版社,2003:221.Li Y,Tu L G.Changbai mountain mushroom[M].Beijing:Science Press,2003:221.
[3]图力古尔.多彩的蘑菇世界:东北亚地区原生态蘑菇图谱[M].上海:上海科学普及出版社,2013:182.Tu L G.Colorful mushroom world:map of native agaricus officinalis in northeast Asia[M].ShangHai:Shanghai Popular Science Press,2013:182.
[4]池玉杰,伊洪伟.木材白腐菌分解木质素的酶系统-锰过氧化物酶、漆酶和木质素过氧化物酶催化分解木质素的机制[J].菌物学报,2007,26(1):153-160.Chi Y J,Yi H W. Lignin degradation mechanisms of ligninolytic enzyme system,manganese peroxidase,laccase and lignin peroxidase,produced by wood white rot fungi[J].Mycosystema,2007,26(1):153-160.
[5]陈兆林,胡大烽,李慧蓉.白腐真菌锰过氧化物酶的研究[J].江苏工业学院学报,2006,18(1):14-17.Chen Z L,Hu D F,Li H R. Study of manganese peroxid of white rot fungi[J]. Journal of Jiangsu Polytechnic University,2006,18(1):14-17.
[6]张连慧,刘卫晓,葛克山,等.变色栓菌产锰过氧化物酶的条件优化[J].微生物学通报,2005,32(5):98-102.Zhang L H,Liu W X,Ge K S,et al.The optmiization of manganese peroxidase production by trametes versicolor[J].Microbiology China,2005,32(5):98-102.
[7]尹亮,谭龙飞.黄孢原毛平革菌生产锰过氧化物酶的发酵条件研究[J].生物技术,2004,14(4):40-42.Yin L,Tan L F.Studies on manganese peroxidases fermentation conditions by phanerochaete chrysosporium[J].Biotechnology,2004,14(4):40-42.
[8]池玉杰,闫洪波.红平菇木质素降解酶系统漆酶、锰过氧化物酶及木质素过氧化物酶的检测[J].林业科学,2009,45(12):154-158.Chi Y J,Yan H B. Detection on laccase,manganese peroxidase and lignin peroxidase in ligninolytic enzymes of Pleurotus djamor[J].Scientia Sincae Sinicae,2009,45(12):154-158.
[9]张玉龙,池玉杰,闫洪波.偏肿栓菌产锰过氧化物酶条件优化[J].林业科学,2011,47(8):88-94.Zhang Y L,Chi Y J,Yan H B. Optimization of ferment conditions of manganese peroxidase produced by Trametes gibbosa[J].Scientia Sincae Sinicae,2011,47(8):88-94.
[10]王佳玲,付时雨,余惠生,等.氮源浓度和芳香化合物对白腐菌产木素降解酶的影响[J].纤维素科学与技术,1997,5(4):25-32.Wang J L,Fu S Y,Yu H S,et al.Effects of nitrogen concentration and aromatic compounds on lignin degrading enzymes produced by white rot fungi[J].Journal of Cellulose Science and Technology,1997,5(4):25-32.
[11]侯红漫,蒋娇娇.白腐菌漆酶的生产及其最佳诱导条件[J].大连轻工业学院学报,2003,22(1):28-31.Hou H M,Jiang J J.Production of laccase from white rot fungi and its optimal inducing conditions[J].Journal of Dalian Polytechnic University,2003,22(1):28-31.
[12]王岁楼,王琼波.漆酶在食品工业中的应用及其产生菌的研究[J].食品科学,2005,26(2):260-263.Wang S L,Wang Q B.Application of laccase in the food industy and its producer[J].Food Science,2005,26(2):260-263.
[13]李鑫,李建科,董攀,等.白腐菌发酵培养及诱导剂对漆酶合成的影响[J].食品工业科技,2013,3(5):160-168.Li X,Li J K,Dong P,et al.White-rot fungus fermentation cultivation and effect of inducer on laccase production[J]. Science and Technology of Food Industry,2013,3(5):160-168.
[14]Feng X Y,Chen H Y,Xue D S,et al.Enhancement of Laccase activity by marine-derived Deuteromycete pestalotiopsis sp. J63with agricultural residues and inducers[J]. Biotechnology and Bioengineering,Chinese Journal of Chemical Engineering,2013,21(10):1182-1189.
[15]王忠燕,于萍,陈艳秋.白耙齿菌最佳培养基配方[J].食用菌学报,2013,20(3):15-17.Wang Z Y,Yu P,Chen Y Q.Optimization of selected nutritional parameters affecting the vegetative growth of Irpex lacteus[J].Acta Edulis Fungi,2013,20(3):15-17.
[16]杨晓宽,杜连祥,路福平.白腐菌产锰过氧化物酶培养基的优化[J].生物技术,2004,14(3):49-50.Yang X K,Du L X,Lu F P.Optimization of medium for manganese peroxidase production by white rot fungi[J].Biotechnology,2004,14(3):49-50.