Department of Food Engineering and Technology, Institute of Chemical Technology, Matunga, Mumbai, Maharashtra, India.
Department of Food Engineering and Technology, Institute of Chemical Technology, Matunga, Mumbai, Maharashtra, India.
J Microbiol Methods. 2024 Nov;226:107050. doi: 10.1016/j.mimet.2024.107050. Epub 2024 Sep 29.
Bile salt hydrolase (BSH), a pivotal enzyme in cholesterol management, holds significant promise in both human and animal subjects. This study investigated the effect of fermentation dynamics in Heyndrickxia coagulans ATCC 7050 and Lactiplantibacillus plantarum ATCC 10012 to enhance BSH production. Cultivation of cultures in MRS and M17 media revealed that MRS medium enhanced BSH production by 235.98 % in H. coagulans ATCC 7050 and 147.37 % in L. plantarum ATCC 10012, compared to M 17 medium. Additionally, varying oxygen concentration levels indicated that H. coagulans ATCC 7050 exhibited its minimum doubling time of 79.8 ± 0.64 min in anaerobic conditions, whereas L.plantarum ATCC 10012 demonstrated its minimum doubling time of 85.5 ± 1.2 min under microaerophilic conditions. However, their highest BSH activity was observed during the stationary phase under anaerobic conditions, yielding 17.14 ± 0.78 U/mL by H. coagulans ATCC 7050 and 19.04 ± 0.81 U/mL by L.plantarum ATCC 10012. Furthermore, it was observed that both organisms did not retain BSH within their cells. BSH activity was assessed using ninhydrin assay that detected free taurine liberated from sodium taurocholate. However, ninhydrin can yield false-positive results owing to its interaction with other free amino acids. To subjugate this limitation, the study introduced a novel and sensitive HPTLC-MS method capable of accurately detecting taurine. By comprehending fermentation dynamics and selecting appropriate conditions, BSH production increased 2.1-fold in both organisms. These findings illuminate critical insights, offering a pathway for novel strategies to enhance the BSH-producing capabilities of these LAB strains.
胆盐水解酶（BSH）是胆固醇管理中的关键酶，在人类和动物研究中都具有很大的应用前景。本研究探讨了凝结芽孢杆菌 ATCC 7050 和植物乳杆菌 ATCC 10012 的发酵动态对提高 BSH 产量的影响。在 MRS 和 M17 培养基中培养发现，与 M17 培养基相比，MRS 培养基使凝结芽孢杆菌 ATCC 7050 的 BSH 产量提高了 235.98%，使植物乳杆菌 ATCC 10012 的 BSH 产量提高了 147.37%。此外，不同的氧浓度水平表明，凝结芽孢杆菌 ATCC 7050 在厌氧条件下的最小倍增时间为 79.8±0.64min，而植物乳杆菌 ATCC 10012 在微需氧条件下的最小倍增时间为 85.5±1.2min。然而，它们的最高 BSH 活性是在厌氧条件下的静止期观察到的，凝结芽孢杆菌 ATCC 7050 产生 17.14±0.78U/mL，植物乳杆菌 ATCC 10012 产生 19.04±0.81U/mL。此外，观察到两种菌都没有将 BSH 保留在细胞内。使用茚三酮法测定 BSH 活性，该方法检测从牛磺胆酸钠中释放的游离牛磺酸。然而，由于其与其他游离氨基酸的相互作用，茚三酮可能会产生假阳性结果。为了克服这一限制，本研究引入了一种新颖而灵敏的 HPTLC-MS 方法，能够准确检测牛磺酸。通过了解发酵动态并选择合适的条件，两种菌的 BSH 产量都提高了 2.1 倍。这些发现提供了重要的见解，为增强这些 LAB 菌株的 BSH 产生能力提供了新的策略途径。
