阿魏侧耳液体发酵及产木质素降解酶培养基的优化
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摘要
以阿魏侧耳NB-1为供试菌株,通过液体发酵培养,测定最佳发酵时间,以生物量、漆酶活力、锰过氧化物酶活力为指标,旨在对培养基中碳源、氮源、生长因子进行优化。结果表明:阿魏侧耳液体发酵最佳培养时间为7 d;4个碳源因素中,可溶性淀粉、蔗糖组生物量极显著高于葡萄糖、麦芽糖组(P<0.01),可溶性淀粉组漆酶活力极显著高于其它3个碳源组(P<0.01),麦芽糖组锰过氧化物酶活力极显著高于其它3个碳源组(P<0.01);4个氮源因素中,牛肉膏、蛋白胨组生物量极显著高于尿素、硫酸铵组(P<0.01),牛肉膏组漆酶活力极显著高于其它3个氮源组(P<0.01),硫酸铵、牛肉膏、尿素组锰过氧化物酶活力极显著高于蛋白胨组(P<0.01);4个生长因子组生物量均极显著优于基础培养基对照组(P<0.01),维生素C组最佳,维生素C组锰过氧化物酶活力极显著高于维生素B6组和基础培养基对照组(P<0.01)。综上所述,阿魏侧耳液体发酵最佳培养时间为7 d,阿魏侧耳液体发酵降解木质素培养基最优碳源为可溶性淀粉,最优氮源为牛肉膏,最优生长因子为维生素C。
Pleuratus ferulae NB-1 was used as donor strain,with liquid fermentation to determine the optimum fermentation time. Besides,this experiment optimized carbon source,nitrogen source and growth factor in medium using biomass,laccase activity and the activity of manganese-dependent peroxidase as index. The results showed that the optimum incubation time of liquid fermentation in Pleuratus ferulae was 7 days. For carbon source,the contents of biomass in soluble starch group and sucrose group were significantly higher than those in glucose group and maltose group( P < 0. 01). The laccase activity in soluble group was significantly higher than that in other groups( P < 0. 01). The highest activity of manganese-dependent peroxidase was maltose group( P < 0. 01). When it came to nitrogen source factors,the contents of biomass in beef extract group and peptone group were remarkably higher than those in urea group and ammonium sulfate group( P <0. 01). Beef extract group' s laccase activity was significantly higher than that in other three groups( P <0. 01). The activity of manganese-dependent peroxidase in peptone group was minimum,and significantly lower than ammonium sulfate group,beef extract group and urea group( P < 0. 01). All the four growth factor group's biomass were remarkably higher than those in control group( P < 0. 01),and vitamin C group was the best.The activity of manganese-dependent peroxidase in vitamin C group was significantly higher than that in vitamin B group and control group( P < 0. 01). It was concluded that the best incubation time of liquid fermentation in Pleuratus ferulae was 7 days,and the optimal carbon source,nitrogen source and growth factor in medium were soluble starch,beef extract and vitamin C respectively.
Pleuratus ferulae NB-1 was used as donor strain,with liquid fermentation to determine the optimum fermentation time. Besides,this experiment optimized carbon source,nitrogen source and growth factor in medium using biomass,laccase activity and the activity of manganese-dependent peroxidase as index. The results showed that the optimum incubation time of liquid fermentation in Pleuratus ferulae was 7 days. For carbon source,the contents of biomass in soluble starch group and sucrose group were significantly higher than those in glucose group and maltose group( P < 0. 01). The laccase activity in soluble group was significantly higher than that in other groups( P < 0. 01). The highest activity of manganese-dependent peroxidase was maltose group( P < 0. 01). When it came to nitrogen source factors,the contents of biomass in beef extract group and peptone group were remarkably higher than those in urea group and ammonium sulfate group( P <0. 01). Beef extract group' s laccase activity was significantly higher than that in other three groups( P <0. 01). The activity of manganese-dependent peroxidase in peptone group was minimum,and significantly lower than ammonium sulfate group,beef extract group and urea group( P < 0. 01). All the four growth factor group's biomass were remarkably higher than those in control group( P < 0. 01),and vitamin C group was the best.The activity of manganese-dependent peroxidase in vitamin C group was significantly higher than that in vitamin B group and control group( P < 0. 01). It was concluded that the best incubation time of liquid fermentation in Pleuratus ferulae was 7 days,and the optimal carbon source,nitrogen source and growth factor in medium were soluble starch,beef extract and vitamin C respectively.
引文
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[19]金晓婷,张志勇,杨栋,等.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.
[20]闫训友,史振霞,唐建国,等.不同氮源对阿魏菇菌丝生长量及多糖含量的影响[J].安徽农业科学,2006,34(20):5206-5207.
[21]崔艳红,韩庆功,常魁珍,等.木质层孔菌产锰过氧化物酶条件的优化及酶学性质研究[J].饲料工业,2012(12):55-59.
[2]王宜磊.侧耳液体培养特性及胞外酶活性研究[J].中国食用菌,2000,19(4):33-35.
[3]李燕荣,周国英,胡清秀,等.食用菌生物降解木质素的研究现状[J].中国食用菌,2009(5):3-6.
[4]AKIN D E,BARTON F E.Rumen microbial attachment and degradation of plant cell walls[J].Fedeation Proceedings,1983,42(1):114-121.
[5]AKIN D E,RIGSBY L L,THEODOROVE M K,et al.Population changes of fibrolytic rumen bacteria in the presence of phenolic acids and plant extracts[J].Animal Feed Science and Technology,1988(19):261-275.
[6]DEVENDRA C.Nutritional ecology of the ruminant[J].Livestock Production Science,1995,43(2):179-180.
[7]王海磊,李宗义.三种重要木质素降解酶研究进展[J].生物学杂志,2003,20(5):9-11,33.
[8]陈友枝.Pleurotus ferulae发酵产漆酶的研究[D].无锡:江南大学,2013.
[9]徐建中,胡开辉,孙淑静,等.不同类型食用菌产木素氧化酶系能力比较与分析[J].江西农业大学学报,2011(2):375-380.
[10]宫志远,于淑芳,曲玲.碳、氮营养对杏鲍菇菌丝生长的影响[J].中国食用菌,2002,24(3):24-26.
[11]KOBAYASHI T,VEN D G,MOO-YOUNG M.Oxygen transfer into mycelial pellets[J].Biotechnology and Bioengineering,1973,15(1):27-45.
[12]胡开辉,刘建忠,孙淑静,等.斑玉覃育种中漆酶转化体系建立的初步研究[J].菌物学报,2010,4(12):56-61.
[13]WANG N,REN K,JIA R,et al.Expression of a fungal manganese peroxidase in Escherichia coli:A comparison between the soluble and refolded enzymes[J].BMC Biotechnology,2016(16):87-103.
[14]何国庆,丁立孝.食品微生物学[M].北京:中国农业大学出版社,2009.
[15]朱妍梅,崔小平.阿魏菇液体培养基的筛选试验[J].食用菌,2006(5):34.
[16]闫训友,祖桂芳,史振霞,等.不同碳源对阿魏菇菌丝生长量及多糖产量的影响[J].食品科技,2007,14(2):38-40.
[17]李永泉,吴炬,杨全珍,等.白阿魏菇深层发酵工艺的研究[J].西北民族大学学报(自然科学版),2003(4):14-18.
[18]张锁峰,柴美清,宋卓琴,等.不同碳源对白灵菇菌丝生长的影响[J].农学学报,2014,4(10):84-87.
[19]金晓婷,张志勇,杨栋,等.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.
[20]闫训友,史振霞,唐建国,等.不同氮源对阿魏菇菌丝生长量及多糖含量的影响[J].安徽农业科学,2006,34(20):5206-5207.
[21]崔艳红,韩庆功,常魁珍,等.木质层孔菌产锰过氧化物酶条件的优化及酶学性质研究[J].饲料工业,2012(12):55-59.