Marine Biotechnology, Fish Nutrition, and Health Division, ICAR-Central Marine Fisheries Research Institute (ICAR-CMFRI), Post Box No. 1603, Ernakulam North P.O., Kochi, 682018, India.
Cochin University of Science and Technology, Cochin, Kerala, India.
Arch Microbiol. 2024 Oct 9;206(11):429. doi: 10.1007/s00203-024-04155-5.
Polyunsaturated fatty acids (PUFA) are vital molecules in the pharmaceutical, medical, and nutritional industries. Exploration of bacterial strains capable of producing significant amounts of PUFAs offers a promising avenue for biotechnological applications and industrial-scale production. However, an extensive screening of several samples from diverse sources is highly needed to identify a potential strain. The present study provides the results of the evaluation of 15 different screening methodologies (including changes in existing protocols in terms of reagent concentration, incubation temperature and time) for identifying PUFA-producing bacteria in comparison to the gold standard method (Gas chromatography-mass spectrometry), for the first time. The results determined the most effective techniques for each critical PUFA, leading to an optimized screening process that saves time and resources. The HO plate assay using 0.5% or 1% HO for 72 & 96 h of incubation at 15 °C consistently outperformed others for finding bacteria containing total nutritionally important long chain-PUFA (LC-PUFA), linoleic acid, and arachidonic acid. Whereas the 2,3,5-triphenyl tetrazolium chloride broth assay at 10-15 °C was the most effective and semiquantitative screening methodology for eicosapentaenoic acid (EPA) and alpha-linolenic acid-containing bacteria. Apart from the methodological perspectives, the study also revealed certain potential strains to be targeted in the ongoing research on PUFA-containing bacteria. Further, the manuscript forms the first report on the presence of docosahexaenoic acid (DHA) in Shewanella decolorationis, EPA in Psychrobacter maritimus and Micrococcus aloeverae, and both EPA and DHA in Arthrobacter rhombi. Altogether, the paper generates several thought-provoking insights on the methodological perspectives and identifies potential PUFA-containing bacteria with practical applications in future bacteria-based PUFA research.
多不饱和脂肪酸（PUFA）是制药、医学和营养工业中至关重要的分子。探索能够大量生产 PUFAs 的细菌菌株为生物技术应用和工业规模生产提供了有前途的途径。然而，需要对来自不同来源的多个样本进行广泛筛选，以鉴定潜在的菌株。本研究首次评估了 15 种不同的筛选方法（包括根据试剂浓度、孵育温度和时间改变现有方案），以鉴定产 PUFAs 的细菌与金标准方法（气相色谱-质谱法）相比。结果确定了每种关键 PUFAs 的最有效技术，从而优化了筛选过程，节省了时间和资源。HO 平板测定法，在 15°C 下孵育 72 和 96 小时，使用 0.5%或 1%HO，对于发现含有总营养长链-PUFA（LC-PUFA）、亚油酸和花生四烯酸的细菌，始终优于其他方法。2,3,5-三苯基氯化四氮唑肉汤测定法在 10-15°C 下是最有效和半定量的筛选方法，适用于含有二十碳五烯酸（EPA）和α-亚麻酸的细菌。除了方法学的角度外，该研究还揭示了某些潜在的菌株，作为正在进行的含 PUFAs 细菌研究的目标。此外，该手稿首次报道了希瓦氏菌属（Shewanella decolorationis）中存在二十二碳六烯酸（DHA）、海冰菌属（Psychrobacter maritimus）和微球菌属（Micrococcus aloeverae）中存在二十碳五烯酸（EPA）以及节杆菌属（Arthrobacter rhombi）中存在 EPA 和 DHA。总之，该论文就方法论的角度提出了一些发人深省的见解，并确定了具有未来基于细菌的 PUFAs 研究实际应用潜力的潜在含 PUFAs 细菌。
