ProBacLab, Advanced Convergence, Handong Global University, Pohang, Gyeongbuk, South Korea.
J Appl Microbiol. 2021 Sep;131(3):1226-1239. doi: 10.1111/jam.15042. Epub 2021 Feb 26.
The objective of this study was to isolate a bacteriocin-producing strain and to characterize the expressed bacteriocin for the control of Listeria monocytogenes with aim of biopreservation application.
Soil samples from a Korean organic farm were subjected to microbiological analysis for isolation of potential bacteriocinogenic LAB, based on a three-level approach, using L. monocytogenes ATCC 15313 as an indicator test micro-organism. From a total of 17 isolates with inhibitory potential, seven were confirmed to be bacteriocin producers. The selected isolates were differentiated based on their morphology, catalase reaction, sugar fermentation profile obtained by API50CHL and by RAPD-PCR generating two unique profiles. One of the isolates, ST110LD, a specific strong producer of anti-Listeria bacteriocins (12 800 AU ml ) was identified as Leuconostoc citreum. The proteinaceous nature of the inhibitory compound produced by Leuc. citreum ST110LD was confirmed through treatment with pepsin and α-chymotrypsin. Bacteriocin activity was observed to be not affected by the presence of milk, NaCl, SDS, Tween 80 or glycerol. Bacteriocin ST110LD effectively inhibited the growth of exponentially growing L. monocytogenes ATCC 15313 during a 10-h incubation period in BHI at 37°C. In addition, this bacteriocin showed specific inhibition of only Listeria spp., but did not inhibit the growth of beneficial cultures included in the microbial test panel for assessment of the spectrum of activity.
Leuconostoc citreum ST110LD was evaluated as safe bacterium strain, producing bacteriocin with high specificity against listerial and enterococcal species. Specificity of producer strain and expressed bacteriocin can be explored in biopreservation of different fermented food products or applied in biotherapy of antibiotic resistant listerial or enterococcal infections.
To the best of our knowledge, this is the first report of bacteriocin produced by Leuc. citreum strain with highly specific antimicrobial activity against Listeria sp. and Enterococcus sp.
本研究的目的是分离产细菌素的菌株,并对表达的细菌素进行特性分析,以期将其应用于李斯特菌的生物防治。
从韩国有机农场的土壤样本中,采用基于三级方法的微生物分析,以李斯特菌 ATCC 15313 为指示测试微生物,分离潜在的产细菌素 LAB。在具有抑制潜力的 17 个分离物中,有 7 个被确认为细菌素产生菌。根据形态学、过氧化氢酶反应、API50CHL 获得的糖发酵谱以及 RAPD-PCR 生成的两种独特图谱对所选分离物进行区分。其中一个分离物 ST110LD 是一种特定的李斯特菌强细菌素产生菌(12 800 AU ml),被鉴定为柠檬明串珠菌。通过用胃蛋白酶和α-糜蛋白酶处理,证实了柠檬明串珠菌 ST110LD 产生的抑制化合物为蛋白质性质。在 37°C 的 BHI 中,10 小时的孵育期内,细菌素 ST110LD 有效地抑制了指数生长期的李斯特菌 ATCC 15313 的生长。此外,该细菌素仅特异性抑制李斯特菌属,但不抑制微生物测试面板中包含的有益培养物的生长,这些培养物用于评估其活性谱。
柠檬明串珠菌 ST110LD 被评估为安全菌株,产生的细菌素对李斯特菌和肠球菌具有高度特异性。产细菌素菌株和表达的细菌素的特异性可在不同发酵食品的生物保鲜中进行探索,或应用于对抗生素耐药李斯特菌或肠球菌感染的生物治疗。
据我们所知,这是首次报道柠檬明串珠菌产生的细菌素对李斯特菌和肠球菌具有高度特异性抗菌活性。