NBRC 3070 植物乳杆菌 BioVector® NBRC 3070 Lactiplantibacillus plantarum

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BioVector® NBRC 3070 植物乳杆菌

菌种名称：植物乳杆菌植物亚种 (Lactiplantibacillus plantarum subsp. plantarum，曾用名：Lactobacillus plantarum / Lactobacillus arabinosus)
菌株保藏编号：NBRC 3070 (等同于 IFO 3070 / ATCC 8014 / DSM 20205 / JCM 1057 / NCIMB 8030)
分类学地位：细菌界、厚壁菌门、芽孢杆菌纲、乳杆菌目、乳杆菌科、乳植物杆菌属
历史与来源：该标准菌株有着清晰的转让历史，最初源自威斯康星大学（由 E. McCoy 分离，编号 17-5），后交由 ATCC 扩增，再经东京大学等途径最终转入日本产品评价技术基盘机构（NITE）生物资源中心（NBRC）保藏。
生物安全级别：1级 (BSL-1 / L1)。该菌株为食品级安全有益益生菌，非致病性，可在常规微生物实验室安全操作。

形态特征：革兰氏阳性菌（Gram-positive），菌体呈直或微弯的杆状，通常呈单个、成对或短链状排列，无芽孢。
呼吸类型：兼性厌氧或微需氧。在体外培养时，更倾向于在无氧/厌氧环境或液体深层静置状态下旺盛生长。
培养基推荐：
- 常规基质：MRS 培养基（de Man, Rogosa and Sharpe medium）。
- 水配方（复苏用）：NBRC 310 号专用复苏液/培养基。
物理培养参数：
- 最佳生长温度：30°C 至 37°C 之间（NBRC 官方推荐维持温度为 30°C）。
- 操作建议：建议使用厌氧罐（Anaerobic jar）进行培养，或者使用经沸水脱气后的 10 mL 液体培养基进行静态深层培养。

冻干管（玻璃安瓿）复苏：
- 用无菌棉签或酒精棉球擦拭安瓿管外壁，在管颈部用砂轮轻划。
- 滴加少量无菌水使管壁局部骤冷裂开，用镊子轻轻敲碎管帽。
- 吸取约 0.5 mL 预热的 MRS 液体培养基注入安瓿管中，温和轻吸几下，使管底的冻干菌块完全溶解并形成均匀的悬液。
- 将溶解后的菌液全部移入含 5–10 mL 新鲜 MRS 液体培养基的试管中。
- 置于 30°C（或 37°C）厌氧或静置培养 18–24 小时，直至液体呈现明显的浑浊状态，证实增殖成功。
平板划线与传代：
- 取增殖好的菌液，在 MRS 固体琼脂平板上进行分区分离划线。
- 置于厌氧培养箱中培养 24–48 小时，待长出边缘整齐、表面光滑、呈乳白色或微灰白色的圆形微凸菌落。
- 挑选单菌落进行后续的实验或甘油冻存（长期保藏建议添加 20%–30% 无菌甘油置于 -80°C 储存）。

维生素与生物活性物质定量生物测定（Bioassay）：该菌株是国际标准的检测指示菌，广泛用于微量分析测定：
- 生物素（Biotin） 测定。
- 烟酸（Nicotinic acid） 测定。
- 泛酸（Pantothenic acid） 测定。
- 对氨基苯甲酸（p-Aminobenzoic acid） 测定。
益生菌微胶囊与肠道递送系统开发（Encapsulation）：由于其良好的肠道耐受潜力，该菌株频繁作为标准对象，用于研究海藻酸钠-芦荟胶基质、油包水/水包油包水（W/O/W）双重乳液微胶囊包裹技术，以提高益生菌在模拟胃酸（SGF/SGJ）和肠液（SIF）中的存活率。
食品发酵与保鲜工艺研究：广泛应用于果汁（如刺果番荔枝汁等发酵）、发酵乳制品以及青贮饲料的发酵动力学、代谢产物分析与天然防腐剂（乳酸菌素）的研发。

Organism Name: Lactiplantibacillus plantarum subsp. plantarum (Former names: Lactobacillus plantarum / Lactobacillus arabinosus)
Strain Catalog Number: NBRC 3070 (Equivalent designations: IFO 3070 = ATCC 8014 = DSM 20205 = JCM 1057 = NCIMB 8030)
Taxonomic Lineage: Bacteria; Bacillati; Lactobacillales; Lactobacillaceae; Lactiplantibacillus
History & Provenance: This reference strain has a well-documented lineage. It was originally isolated at the University of Wisconsin (by E. McCoy, designated as strain 17-5), deposited into the ATCC, transferred through the University of Tokyo, and ultimately cataloged into the NBRC collection managed by NITE.
Biosafety Level: Biosafety Level 1 (BSL-1 / L1).Classified as a safe, food-grade probiotic bacterium with zero human pathogenicity, suitable for handling under standard laboratory infrastructure.

Morphology: Gram-positive, non-spore-forming rod-shaped bacterium. Cells appear straight or slightly curved with rounded ends, presenting individually, in pairs, or arranged in short chains.
Oxygen Relationship: Facultative anaerobe / microaerophilic.It demonstrates optimal metabolic kinetics and vigorous biomass yield when cultured statically or under strictly anaerobic conditions.
Recommended Cultivation Media:
- Standard Choice: MRS (de Man, Rogosa and Sharpe) medium.
- Rehydration Fluid: NBRC Medium No. 310 formulation.
Physical Incubation Thresholds:
- Optimal Incubation Temperature: Tolerable from 30°C to 37°C, with the NBRC official validation parameter set firmly at 30°C.
- Atmospheric Suggestions: Cultivate using an anaerobic jar or incubate statically in a 10 mL deep liquid column previously degassed in boiling water.

Rehydration of L-Dried Specimens (Glass Ampoules):
- Disinfect the outer surface of the glass ampoule using an alcohol wipe and score around the neck section with a sharp file.
- Apply a localized cold stimulus to create a clean fracture line, then carefully open the top of the vial.
- Aseptically introduce approximately 0.5 mL of pre-warmed MRS broth into the ampoule, pipetting up and down gently to completely dissolve the lyophilized bacterial pellet.
- Transfer the entirety of this rehydrated suspension into a culture tube filled with 5–10 mL of fresh MRS liquid medium.
- Incubate at 30°C (or 37°C) under anaerobic or static conditions for 18–24 hours until noticeable, uniform turbidity indicates successful vegetative growth.
Plating and Long-Term Maintenance:
- Streak a loopful of the turbid suspension onto solid MRS agar plates to obtain isolated colonies.
- Incubate anaerobically for 24–48 hours to yield circular, smooth, convex, opaque white to cream-colored distinct colonies.
- Pick individual colonies for scaling up or preserve them long-term at -80°C by supplementing clean log-phase cultures with 20%–30% sterile glycerol.