KMITL Expo 2026 LogoKMITL 66th Anniversary Logo

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

Other Innovations

Cracking the PM2.5 Code

คณะวิทยาศาสตร์

Cracking the PM2.5 Code

Air pollution, particularly PM2.5, is a major environmental and public health concern in Bangkok. Instead of predicting PM2.5 levels, this project aims to identify the most significant factors influencing PM2.5 concentration. By analyzing historical air quality, weather, and other environmental data, we will determine which variables—such as temperature, humidity, wind speed, or other pollutants—have the greatest impact on PM2.5 fluctuations.

Read more
Farm Life Agricultural Design

คณะเทคโนโลยีการเกษตร

Farm Life Agricultural Design

This project presents a design and management approach for agricultural land in Kanchanaburi Province. The case study area is situated in Wangdong Subdistrict, Mueang Kanchanaburi District, covering an area of approximately 18 rai (7.2 acres). As the user seeks a simplified lifestyle in the countryside, surrounded by nature, the design aligns with this vision of simplicity and sustainability. The land is systematically allocated to optimize the benefits for both daily living and agricultural industry development. The crop cultivation zones are designed to suit the local climate and plant varieties, ensuring high-quality yields for continuous utilization. Meanwhile, the livestock zones are clearly delineated to maintain balance and organization. This approach not only ensures food security and income generation but also promotes a lifestyle that harmonizes with nature, minimizes environmental impact, and supports the long-term development of an efficient and eco-friendly agricultural industry. Comprehensive attention is given to the positioning of various zones, considering wind direction and sunlight exposure. Additionally, the design undergoes a rigorous drafting and review process to ensure the optimal outcomes for the land's utilization.

Read more
Synergistic antibacterial activity of human salivary peptide Histatin-5 and Phytosphingosine against Streptococcus mutans biofilm

คณะทันตแพทยศาสตร์

Synergistic antibacterial activity of human salivary peptide Histatin-5 and Phytosphingosine against Streptococcus mutans biofilm

Objective or Background: Dental caries is still one of the most significant dental problems worldwide, with prevalence rates up to 90% among children and adults. Cariogenic bacteria, especially Streptococcus mutans, is the primary microorganism involved in the pathogenesis through carbohydrate metabolism and biofilm formation, which are challenging to eradicate. Histatin-5 (HST-5), a human salivary antimicrobial peptide, has demonstrated antimicrobial activity against various fungal and bacterial pathogens. Phytosphingosine (PHS), an endogenous bioactive sphingolipid found in fungi, plants, and humans, also shows antimicrobial properties. This study aimed to evaluate the killing activity of HST-5 alone and in combination with PHS against S. mutans under biofilm-stimulating conditions. Materials and Methods: Antimicrobial activity against a planktonic culture of S. mutans was evaluated using a time-kill assay, and biofilm-forming capacity was confirmed by crystal violet staining assay. The killing ability against 24h pre-formed biofilm was determined using Transferable Solid Phase (TSP) pin lid model. Synergistic activity between HST-5 and PHS was evaluated using the checkerboard technique. Additionally, the cytotoxicity of the tested agent on human gingival fibroblast cells (hGFs) was assessed after 1 h of incubation using an MTT assay. Results: A time-kill assay revealed that both HST-5 and PHS exhibit time- and concentration-dependent activity against the planktonic form of S. mutans. PHS achieved over 90% killing activity within 15 min at 5 μg/ml, whereas HST-5 required 30 min to reach 90% killing at 20 μM. The biofilm formation capacity of S. mutans was confirmed. The inhibitory concentrations (IC50) of HST-5 and PHS against S. mutans biofilm were 25 μM and 13.5 μg/ml, respectively. A synergistic interaction between HST-5 and PHS, with IC50 values reduced by 8-fold and 16-fold, respectively. No cytotoxic effects were observed in hGFs cells at the concentration of the synergistic interaction. Conclusions: Therefore, the combination of HST-5 and PHS may enhance the effectiveness of anti-infective agents against S. mutans biofilm, potentially preventing the development of dental caries.

Read more