环境温度和酸碱变化适应性广的纤维素降解菌复合系筛选  

Screening of Cellulose Degradation Bacteria Composite System withWide Adaptability of Temperature and Acid Base Changes in Surroundings

作　　者：肖艳萍[1,2,3] 张仕颖 包立[1,2] 赵丹丹 张乃明[1,2] XIAO Yan-ping;ZHANG Shi-ying;BAO Li;ZHAO Dan-dan;ZHANG Nai-ming(College of Plant Protection,Yunnan Agricultural University,Yunnan Kunming 650201,China;Yunnan Engineering Laboratory of Soil Fertility and Pollution Remediation,Yunnan Soil Resources Protection and Utilization Innovation Team,Yunnan Kunming 650201,China;Economic and Trade School in Yunnan Province,Yunnan Kunming 650204,China;Institute of Agricultural Economics and Information,Yunnan Academy of Agricultural Sciences,Yunnan Kunming 650231,China)

机构地区：[1]云南农业大学植物保护学院,云南昆明650201 [2]云南省土壤培肥与污染修复工程实验室/云南省土壤资源利用与保护创新团队,云南昆明650201 [3]云南省贸易经济学校,云南昆明650204 [4]云南省农业科学院农业经济与信息研究所,云南昆明650231

出　　处：《西南农业学报》2020年第5期1019-1027,共9页Southwest China Journal of Agricultural Sciences

基　　金：科技创新人才计划(2015HC018);水污染防治重大专项(2012ZX01702-003);对外科技合作计划-院士专家工作站(2015IC022)。

摘　　要：【目的】外源微生物在生物堆肥过程中的作用是否稳定对生物堆肥意义重大,本研究旨在获得具有较好环境耐受性的复合微生物菌,增强菌剂在生物堆肥中的竞争力和实用效果。【方法】将前期从自然环境中分离到的具有纤维素分解能力的20株菌,构建25组复合微生物菌剂,经过拮抗关系试验筛选到21组复合微生物剂,并通过液体发酵培养,采用DNS法测定复合菌的纤维素酶活:包括滤纸酶活(FPA)和不同环境条件下(温度、酸碱)的羧甲基纤维素钠酶活(CMC酶活)。【结果】筛选得到的复合微生物CM17,该菌系在35、45、55℃,pH值6、7.5、10,都能正常分解纤维素,具有良好的环境耐受性。16SrDNA分子鉴定表明,CM17复合菌剂中包含有链格孢菌(Alternaria alternate)、链霉菌(Streptomyces sp.)、厄氏菌(Oerskovia sp.)和短梗霉菌(Aureobasidium sp.)等。CM17复合菌剂的纤维素酶活远远比单菌株强,当4种菌的混合体积配比为1∶1∶1∶1时,其分解纤维素的酶活功能表现最佳。【结论】CM17复合菌剂具有较强的环境耐受性,对复杂环境下生物堆肥具有一定的理论和应用价值。【Objective】Whether the effect of exogenous microorganism was stable was a great significance to biological composting.The present paper aimed to obtain compound microbial inoculum with better environmental tolerance and enhance the competitiveness and effect of the microorganism bacteria in biological composting.【Method】25 groups of compound microbes were constructed from 20 strains with cellulose decomposing ability isolated from the natural environment in the previous research work.21 groups of compound microbes were screened out through the antagonistic relationship test,and the cellulase activities of 21 groups were determined by the method of liquid fermentation and DNS,which included filter paper enzyme activity(FPase)and carboxymethyl cellulose sodium enzyme activity(CMCase)under different temperature conditions(35,45 and 55℃)and different acid and alkali conditions(pH value 6,7.5 and 10).【Result】Finally,CM17 with wide temperature adaptability and acid-base adaptability was screened out,and the compound bacteria could decompose cellulose normally within the temperature from 35 to 55℃and the pH value 6 to 10.16SrDNA molecular identification showed that the 4 strains of CM17 were Alternaria alternate,Streptomyces sp.,Oerskovia sp.and Aureobasidium sp..CM17 had better cellulase activity than single strain,and when the ratio of 4 bacteria was 1∶1∶1∶1,the compound had the best cellulolytic enzyme activity.【Conclusion】CM17 was stable and had better environmental tolerance.This study had certain theoretical and applied value for biological composting in complex environment.

关 键 词：生物堆肥 复合微生物 筛选 酶活 

分 类 号：O636[理学—高分子化学]