摘要
为深入研究堆肥过程中主要微生物的功能,以番茄秸秆为原料,对其反应器堆肥过程进行研究。通过反应器自动温度传感器记录堆肥温度,利用分光光度法测定堆肥过程中植物生物质降解相关酶活力,并对酶活力最高时期的典型代表性样品进行高通量Orbitrap质谱鉴定。结果表明,反应器堆肥共持续35天,其中50℃以上的高温期10天以上,发酵至第7天时,内切纤维素酶和木聚糖酶活力最高。对该时期样品进行质谱鉴定发现,Thermobifida是主要的功能细菌,且是唯一的纤维素降解者,产生糖苷水解酶(GH)5家族和GH9家族的5种内切纤维素酶和3种GH48家族的外切纤维素酶。Planifilum产生GH3家族的β-葡萄糖苷酶,与Thermobifida一起将纤维素降解成葡萄糖。Thermobifida、Planifilum、Saccharomonospora、Aspergillus和Thermomyces共同参与木聚糖的降解,前三者为功能细菌,主要分泌GH10家族的木聚糖酶,后二者为功能真菌,主要分泌GH11家族的木聚糖酶。
To further identify functions of key microorganisms in composting,the reactor composting process using tomato straw as raw material was studied.The compost temperature was auto-monitored by the temperature sensor,and the spectrophotometric method was applied in determining activities of the lignocellulosic enzymes.The sample with the highest activity was selected as the typical representative for further identification by high-throughput Orbitrap-LC-MS/MS.The results showed that,the thermophilic phase with temperature>50℃lasted for more than 10 days during the whole period of 35 days,and the activities of both endoglucanase and xylanase peaked on the 7th day.The Orbitrap analysis revealed that Thermobifida was the only cellulose degrader secreting five endoglucanases including members of glycoside hydrolase(GH)family 5 and GH family 9,and three cellobiohydrolases from GH48.Planifilum cooperated with Thermobifida in cellulose degrading to glucose by secretingβ-glucosidase from GH family 3.Thermobifida,Planifilum,Saccharomonospora,Aspergillus and Thermomyces participated in xylan degrading.Xylanases produced by the former three bacteria were members of GH10,and those secreted by the latter two fungi were from GH11.
作者
张小梅
咸洪泉
李雅华
陈清华
Zhang Xiaomei;Xian Hongquan;Li Yahua;Chen Qinghua(College of Resources and Environment,Qingdao Agricultural University,Qingdao 266109,China;Qingdao Engineering Research Center for Rural Environment,Qingdao 266109,China;College of Life Sciences,Qingdao Agricultural University,Qingdao 266109,China)
出处
《山东农业科学》
2020年第4期73-78,共6页
Shandong Agricultural Sciences
基金
国家自然科学基金项目(31600084)
山东省高等学校精品课程(2013BK209)
青岛农业大学名校工程课程建设项目(XJG2013138,XDSJP2013031)
青岛农业大学实验技术研究课题(SYJK15-24,SYJK16-25)。
关键词
堆肥
糖苷水解酶
功能微生物
高通量质谱
Composting
Glycoside hydrolase
Functional microorganism
High-throughput mass spec-trometer
