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纤维素降解菌株的分离与筛选
赖国栋, 秦长生, 赵丹阳, 邱华龙, 杨华, 田龙艳, 练涛, 徐金柱
广东省林业科学研究院
摘要:
从实验室已有的木腐菌与土壤中分离获得的菌株中筛选出具有较高纤维素降解能力的菌株。 利用羧甲基纤维素钠为唯一碳源的选择培养,利用刚果红染色进行初筛,选择透明圈与菌落直径比值较 大的菌落进行酶活测定再次进行复筛。同时,利用分子学手段对高效产胞外纤维素酶的菌株进行鉴定。 最终,筛选出 4 株高效产胞外纤维素酶的菌株,其中菌株 HF-3 在 28 ℃、 180 r/min 条件下发酵培养 5 d 后,胞外纤维素酶活性最高,高达 67.772 U/mL,经鉴定为密褐褶孔菌 Gloeophyllum trabeum;另外 Y5、 Y6、 Y2 共 3 株菌株酶活较高,分别为 37.714 U/mL、 30.694 U/mL、 25.336 U/mL,属于链霉菌,依次鉴 定为浅灰白链霉菌 Streptomyces griseoloalbus、丝状链霉菌 Streptomyces filamentosuss 和威德摩尔链霉菌 Streptomyces wedmorensis。试验筛选得到 HF-3、 Y5、 Y6 以及 Y2 菌株具有较高纤维素降解能力,可为复 合菌系的研制提供材料。
关键词:  纤维素  纤维素降解菌  鉴定  筛选
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基金项目:广东省林业科技计划项目(编号:2019-16)
Isolation and Screening of Cellulose Degrading Strains
laiguodong, qinchangsheng, zhaodanyang, qiuhualong, yanghua, tianlongyan, liantao, xujinzhu
Guangdong Academy of Forestry
Abstract:
Screening strains with high cellulose degradation ability from wood rotting fungi in laboratory and humus layer available.Sodium carboxymethyl cellulose was used as the sole carbon source for selective culture, and Congo red staining was used for primary screening. Colonies with larger ratio of transparent circle to colony diameter were selected for enzyme activity determination for re-screening. Finally, strains with high enzyme activity were identified by molecular method. Four strains with strong cellulose-degrading ability were screened out from the existing wood rotting fungi and strains from humus layer. Among them, HF-3 strain had CMC enzyme activity of 67.772 U/mL after 5 days of fermentation at 28 oC for 180 r/min, which was identified as Gloeophyllum trabeum. The other three strains with higher enzyme activities were identified as Streptomyces griseoloalbus, Streptomyces filamentosus, and Streptomyces wedmorensis, which were 37.714,30.694,and 25.336 U/mL, respectively. In this experiment, HF-3, Y5, Y6, and Y2 strains showed high cellulose degradation ability, which could provide materials for the development of composite strains. The results of this study are of great significance for accelerating the decomposition of green garbage.
Key words:  cellulase  fungi  identification  screen