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Genomic analysis, Bacteriocin dynamic production of putatively novel bacteriocin DCR3-2 inhibiting Listeria monocytogenes synthesized by Lactococcus lactis subsp. hordinae DCR3-2 isolated from Thai pickled crab (Pu-dong)

Abstract

Listeriosis is a severe foodborne illness characterized by a fatality rate exceeding 30%, attributed to the pathogen Listeria monocytogenes. This study evaluated 160 lactic acid bacteria (LAB) isolated from Thai pickled crabs for their potential as agents against L. monocytogenes and for their probiotic properties and probiogenomic characteristics. Among these strains, strain DRC3-2 exhibited activity through the synthesis of bacteriocin DRC3-2, which significantly inhibited L. monocytogenes ATCC 19115 in spot-on-lawn assays. Phenotypic and whole-genome analyses revealed that strain DRC3-2 thrived in environments with 2-6% NaCl, pH values ranging from 3 to 9, and temperatures between 25 and 45°C. Based on average nucleotide identity (ANI) and digital DNA‒DNA hybridization (dDDH) values, strain DRC3-2 was taxonomically classified as Lactococcus lactis subsp. hordinae. The production of bacteriocin DRC3-2 peaked during the late stationary phase, following its synthesis in the early exponential growth phase. BAGEL4 analysis identified the putative novel bacteriocin DRC3-2 as lactococcin A and B, with respective bit-scores of 40.05 and 36.58. In silico safety assessments confirmed the nonpathogenic nature of strain DRC3-2 in humans, highlighting its absence of antibiotic resistance genes. Finally, this investigation underscores the novel bacteriocin DRC3-2 for application in the prevention and treatment of L. monocytogenes infections.

Objective

Listeria monocytogenes เป็นเชื้อก่อโรคที่มักพบในอาหารที่มีการปนเปื้อน สามารถอยู่รอดในสภาวะที่รุนแรง เช่น อุณหภูมิต่ำ ความเป็นกรดสูง ความเข้มข้นของเกลือสูง ซึ่งทำให้มีความเสี่ยงอย่างมากในระบบย่อยอาหารของมนุษย์ ดังนั้นจึงศึกษาแบคทีเรียกรดแลคติกที่มีความสำคัญในอุตสาหกรรมอาหารและยาเนื่องจากมีคุณสมบัติเป็นโพรไบโอติก และแบคทีเรียกรดแลคติกบางชนิดที่สามารถผลิตแบคเทอริโอซินหรือเปปไทด์ต้านจุลชีพที่มีฤทธิ์ทางชีวภาพ และแบคเทอริโอซินที่มีประสิทธิภาพในการยับยั้งแบคทีเรียก่อโรค ซึ่งถือว่าเป็นทางเลือกที่น่าสนใจแทนการใช้ยาปฎิชีวนะแบบดั้งเดิม เนื่องจากช่วยลดการพัฒนาการดื้อยาของแบคทีเรีย นอกจากนี้แบคเทอริโอซินยังมีความเสถียรสูง สามารถทนต่ออุณหภูมิที่รุนแรงและสภาวะเป็นกรดได้ อีกทั้งยังไม่เป็นพิษต่อเซลล์มนุษย์ การศึกษานี้จึงมุ่งหมายเพื่อศึกษาแบคเทอริโอซินชนิดใหม่จาก Lactococcus lactis subsp. hordinae DCR3-2 ซึ่งแยกได้จากปูดอง เพื่อประเมินฤทธิ์การต้านจุลชีพของแบคทีเรียชนิดนี้ และการวิเคราะห์ทางbioinformatics เพื่อยืนยันว่าสายพันธุ์ DCR3-2 มีความปลอดภัยสำหรับมนุษย์และมีคุณสมบัติโพรไบโอติก

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