亮斑扁角水虻卵共生菌Bacillus velezensis的鉴定和产酶特征及其对幼虫分解餐厨垃圾效率的影响  

Identification and enzyme profiling of black soldier fly egg commensal Bacillus velezensis and its effect on food waste bioconversion

作　　者：陈想 赵思捷 梁婷婷 张继冉 陈红歌 杨森 Xiang Chen;Sijie Zhao;Tingting Liang;Jiran Zhang;Hongge Chen;Sen Yang(Key Laboratory of Microbial Engineering,Ministry of Agriculture,College of Life Sciences of Henan Agricultural University,Zhengzhou 450002,Henan Province,China)

机构地区：[1]河南农业大学生命科学学院,农业部农业微生物酶工程重点实验室,河南郑州450002

出　　处：《微生物学报》2021年第7期2121-2135,共15页Acta Microbiologica Sinica

基　　金：河南省重点研发与推广专项(科技攻关192102310240)。

摘　　要：【目的】从亮斑扁角水虻(Hermetia illucens L.)卵表筛选得到一株产多种酶的卵表共生菌,对该菌进行鉴定,并探究其最适生长条件、产酶特性及其对幼虫分解餐厨垃圾效率的影响。【方法】通过多种选择性培养基筛选得到产多种酶的菌株。通过单因素实验方法确定其最适生长条件、产酶特性及其对幼虫分解餐厨垃圾效率的影响。【结果】经过形态学观察、生理生化鉴定和16S rDNA序列分析,将该株亮斑扁角水虻卵表共生菌命名为贝莱斯芽胞杆菌(Bacillus velezensis EEAM 10B)。最适摇瓶培养条件为:40℃,200 r/min,pH 7.0,酵母浸粉10 g/L,葡萄糖10 g/L,培养16 h活菌数达3.1×10^(9) CFU/mL。进入稳定期后开始形成单端生芽胞,24 h后芽胞形成率95.8%。使用产酶筛选培养基培养结果表明:B.velezensis EEAM 10B菌株产木聚糖酶活性最强,其次是蛋白酶、纤维素酶、果胶酶、淀粉酶和植酸酶。按照1×10^(6) CFU/g用量添加B.velezensis EEAM 10B芽胞制剂到餐厨垃圾中饲养亮斑扁角水虻,B.velezensis EEAM 10B芽胞菌剂能够显著(P<0.05)提高亮斑扁角水虻幼虫对灭菌和非灭菌餐厨垃圾的转化效率,分别为13.4%和13.54%,但物料减少率没有显著差异(P>0.05);显著提高灭菌餐厨垃圾中幼虫存活率至95%,提高非灭菌餐厨垃圾饲养幼虫的预蛹单重0.1437 g/只,化蛹率92.57%。【结论】B.velezensis EEAM 10B菌株能够产多种酶,且在亮斑扁角水虻处理餐厨废弃物中有潜在应用价值。[Objective]A strain producing multiple enzymes was isolated from the egg surface of Hermetia illucens L.Further,the optimal growth conditions,enzyme-producing characteristics,and the effects of the strain on the food waste decomposition efficiency of Hermetia illucens L.were explored.[Methods]Enzyme-producing strains were obtained from various selective media.The optimal growth conditions,enzyme-producing characteristics,and the effects of it on the food waste treatment decomposition efficiency of Hermetia illucens L.were determined by the single-factor experiment.[Results]In this study,a Hermetia illucens L.egg-surface commensal bacterium was named as Bacillus velezensis EEAM 10B by morphological observation,biophysiological and biochemical tests,and 16S rDNA sequence analysis.The bacteria number was 3.1×10^(9) CFU/mL under the optimum growth conditions:40°C,200 r/min,pH 7.0,10 g/L yeast extract,10 g/L glucose for 16 h.Approximately 95.8%single-terminal spores were formed after 24 h cultivation.Enzymes producing results are as follows:xylanase activity was the highest,followed by the protease,cellulase,pectinase,amylase,and phytase.Adding the Bacillus velezensis EEAM 10B spores to the food waste at a dosage of 1×10^(6) CFU/g significantly(P<0.05)improved the food conversion efficiency of black soldier fly larvae in both sterilized and non-sterilized food waste groups,reaching 13.4%and 13.54%,respectively,whereas the mass reduction rates did not present differences(P>0.05).Furthermore,in the sterilized food waste group,the survival rate of larvae and the prepupae weight were significantly increased to 95%and 0.1437 g per one,while the pupation rate reached 92.57%in the non-sterilized food waste group.[Conclusion]B.velezensis EEAM 10B strain can produce a variety of enzymes,and it has potential application value in the treatment of food waste with the black soldier fly.

关 键 词：亮斑扁角水虻 卵表共生微生物 贝莱斯芽胞杆菌 产酶特征 餐厨垃圾 

分 类 号：S899.9[农业科学—特种经济动物饲养]