CAJ | 학술논문

本文在对峙法验证贝莱斯芽孢杆菌B. velezensis YB15具抑菌作用的基础上，用透明圈法、DNS法研究其产β-葡聚糖酶特性，利用对峙法验证该酶抑菌作用，通过PCR法获得目的基因，分析克隆序列并预测其蛋白质结构与功能。结果表明，该菌株对多种病原真菌有拮抗作用，杨树紫纹羽病菌拮抗带达11.0 mm，该菌提取的葡聚糖酶粗酶液对杨树紫纹羽病菌抑菌带为10.6 mm，说明葡聚糖酶在菌株YB15抑菌中有重要作用。不同接种方法影响菌株YB15葡聚糖酶水解透明圈形成，点种法水解圈与菌落直径之比在72 h可达14.1，效果最好。克隆所得菌株YB15葡聚糖酶基因命名为Bglu1，该基因序列长732 bp，编码243个氨基酸，此酶蛋白氨基酸序列与解淀粉芽孢杆菌TB2β-1,3-1,4-葡聚糖酶同源性较高，属糖基水解酶16家族，N端疏水区存在信号肽并具跨膜区域，推测其为分泌蛋白。
본문재대치법험증패래사아포간균B. velezensis YB15구억균작용적기출상，용투명권법、DNS법연구기산β-포취당매특성，이용대치법험증해매억균작용，통과PCR법획득목적기인，분석극륭서렬병예측기단백질결구여공능。결과표명，해균주대다충병원진균유길항작용，양수자문우병균길항대체11.0 mm，해균제취적포취당매조매액대양수자문우병균억균대위10.6 mm，설명포취당매재균주YB15억균중유중요작용。불동접충방법영향균주YB15포취당매수해투명권형성，점충법수해권여균락직경지비재72 h가체14.1，효과최호。극륭소득균주YB15포취당매기인명명위Bglu1，해기인서렬장732 bp，편마243개안기산，차매단백안기산서렬여해정분아포간균TB2β-1,3-1,4-포취당매동원성교고，속당기수해매16가족，N단소수구존재신호태병구과막구역，추측기위분비단백。
On the basis of fungus-inhibitory activity tested with dual culture method, authors studied characteristics of β-glucanase produced by B. velezensis YB15. Results showed that the strain played an antagonistic role against many pathogenic fungi. Width of inhibition zone against Helicobasidium purpureum was 11.0 and 10.6 mm by YB15 and itsβ-glucanase crude enzyme, respectively. Different inoculation methods could impose impacts on the formation of the hydrolytic transparent circle of the glucanase of YB15. The proportion of hydrolytic circle and colony diameter by dibbling could be up to 14.1 at 72 h. The cloned gene of YB15β-glucanase named as Bglu1 was 732 bp in length and encoded 243 amino acids. The protein showed high homology with β-1,3-1,4-glucanase of B. amyloliquefaciens TB2 belonging to glycosyl hydrolase-16 family. The N-terminal hydrophobic domain had signal peptide and transmembrance domain.