生物技术通报 ›› 2021, Vol. 37 ›› Issue (11): 158-165.doi: 10.13560/j.cnki.biotech.bull.1985.2021-1120

• 食用菌生物技术专题(专题主编: 黄晨阳) • 上一篇    下一篇

香菇菌渣高效纤维素降解菌的筛选及产酶优化

陶治东1(), 何艳慧1, 邓子禾2, 孙琳琳1, 武占省1()   

  1. 1.西安市纺织化工助剂重点实验室 西安工程大学环境与化学工程学院,西安 710048
    2.石河子大学化学化工学院,石河子 832003
  • 收稿日期:2021-08-31 出版日期:2021-11-26 发布日期:2021-12-03
  • 作者简介:陶治东,男,硕士,研究方向:固体废物资源化;E-mail: 3188937343@qq.com
  • 基金资助:
    陕西省教育厅科研计划项目(20JC014)

Screening of High-efficiency Cellulose-degrading Microorganism from Spent Lentinula edodes Substrate and Optimization of Its Enzyme Production

TAO Zhi-dong1(), HE Yan-hui1, DENG Zi-he2, SUN Lin-lin1, WU Zhan-sheng1()   

  1. 1. Xi’an Key Laboratory of Textile Chemical Engineering Auxiliaries,School of Environmental and Chemical Engineering,Xi’an Polytechnic University,Xi’an 710048
    2. School of Chemistry and Chemical Engineering,Shihezi University,Shihezi 832003
  • Received:2021-08-31 Published:2021-11-26 Online:2021-12-03

摘要:

利用香菇菌渣开发生物有机肥目前是解决废弃菌渣资源化的主要趋势,为了更好地腐熟香菇菌渣,从废弃香菇菌渣中筛选出1株高效纤维素降解菌株DGW1,并优化产纤维素酶条件及评估降解菌渣能力,以期为后续利用香菇菌渣进行堆肥发酵生产生物有机肥奠定基础。采用CMC-Na平板法和刚果红染色法初步筛选,结合滤纸条崩解试验和纤维素酶活性测定进行复筛,并通过形态学观察和18S rDNA分子生物学鉴定菌株,优化菌株DGW1的产纤维素降解酶条件。结果表明:获得的高效纤维素降解菌株DGW1鉴定为里氏木霉,7 d内可将滤纸条降解为糊状。菌株DGW1最佳产酶条件为温度30℃、时间4 d,滤纸酶的酶活(FPase)为112.71 U/g,并在pH 4-7都能达到较高酶活且不低于96 U/g。菌株DGW1对香菇菌渣的降解率为31.14%(14 d)。因此,获得的菌株DGW1能够高效降解香菇菌渣,其纤维素酶活较高,对香菇菌渣发酵堆肥,制备生物有机肥具有较大应用潜力。

关键词: 香菇菌渣, 里氏木霉DGW1, 酶活力, 产酶条件

Abstract:

Utilizing spent Lentinula edodes substrate to develop organic fertilizer is major trend of converting waste bacterial residue into resources. In order to compost the spent L. edodes substrate,a high efficient cellulose-degrading strain DGW1 was isolated from the spent L. edodes substrate,the conditions of cellulase production were optimized,and the degradation capacity of the spent L. edodes substrate was evaluated,which lays a foundation for the subsequent composting fermentation of spent L. edodes substrate to produce bio-organic fertilizer. The strains were preliminarily screened by CMC-Na culturing method and Congo red dyeing and re-screened in combination with filter paper disintegration test and cellulase activity test. The strain was identified by morphological observation and 18S rDNA molecular biology,and the conditions of producing cellulose-degrading enzyme by the strain DGW1 were optimized. The results showed that the highly cellulolytic strain DGW1 was identified as Trichoderma reesei. The filter paper strips were degraded to paste in the 7 d. The optimal conditions of the strain DGW1 were temperature 30℃,time 4 d,the filter paper activity(FPase)was 112.71 U/g,and the high enzyme activity at pH 4-7 was not lower than 96 U/g. The degradation rate of the strain DGW1 on spent L. edodes substrate was 31.14%(14 d). Therefore,the strain DGW1 can effectively degrade spent L. edodes substrate,and its cellulase activity was high,which has great application potential for composting of spent L. edodes substrate and preparation of bio-organic fertilizer.

Key words: spent Lentinula edodes substrate, Trichoderma reesei DGW1, enzyme activity, enzyme production condition