NBRC 3070 植物乳桿菌 BioVector? NBRC 3070 Lactiplantibacillus plantarum
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BioVector? NBRC 3070 植物乳桿菌
BioVector? NBRC 3070 Lactiplantibacillus plantarum
第一部分:中文說明
一、 產(chǎn)品基本信息與微生物學(xué)背景
菌種名稱:植物乳桿菌植物亞種 (Lactiplantibacillus plantarum subsp. plantarum,曾用名:Lactobacillus plantarum / Lactobacillus arabinosus)
菌株保藏編號(hào):NBRC 3070 (等同于 IFO 3070 / ATCC 8014 / DSM 20205 / JCM 1057 / NCIMB 8030)
分類學(xué)地位:細(xì)菌界、厚壁菌門、芽孢桿菌綱、乳桿菌目、乳桿菌科、乳植物桿菌屬
歷史與來源:該標(biāo)準(zhǔn)菌株有著清晰的轉(zhuǎn)讓歷史,最初源自威斯康星大學(xué)(由 E. McCoy 分離,編號(hào) 17-5),后交由 ATCC 擴(kuò)增,再經(jīng)東京大學(xué)等途徑最終轉(zhuǎn)入日本產(chǎn)品評(píng)價(jià)技術(shù)基盤機(jī)構(gòu)(NITE)生物資源中心(NBRC)保藏。
生物安全級(jí)別:1級(jí) (BSL-1 / L1)。
該菌株為食品級(jí)安全有益益生菌,非致病性,可在常規(guī)微生物實(shí)驗(yàn)室安全操作 。
二、 菌體形態(tài)與生理生化特征
形態(tài)特征:革蘭氏陽性菌(Gram-positive),菌體呈直或微彎的桿狀,通常呈單個(gè)、成對或短鏈狀排列,無芽孢。
呼吸類型:兼性厭氧或微需氧。
在體外培養(yǎng)時(shí),更傾向于在無氧/厭氧環(huán)境或液體深層靜置狀態(tài)下旺盛生長。 培養(yǎng)基推薦:
常規(guī)基質(zhì):MRS 培養(yǎng)基(de Man, Rogosa and Sharpe medium)。
水配方(復(fù)蘇用):NBRC 310 號(hào)專用復(fù)蘇液/培養(yǎng)基。
物理培養(yǎng)參數(shù):
最佳生長溫度:30°C 至 37°C 之間(NBRC 官方推薦維持溫度為 30°C)。
操作建議:建議使用厭氧罐(Anaerobic jar)進(jìn)行培養(yǎng),或者使用經(jīng)沸水脫氣后的 10 mL 液體培養(yǎng)基進(jìn)行靜態(tài)深層培養(yǎng)。
三、 菌株復(fù)蘇與傳代標(biāo)準(zhǔn)步驟
凍干管(玻璃安瓿)復(fù)蘇:
用無菌棉簽或酒精棉球擦拭安瓿管外壁,在管頸部用砂輪輕劃。
滴加少量無菌水使管壁局部驟冷裂開,用鑷子輕輕敲碎管帽。
吸取約 0.5 mL 預(yù)熱的 MRS 液體培養(yǎng)基注入安瓿管中,溫和輕吸幾下,使管底的凍干菌塊完全溶解并形成均勻的懸液。
將溶解后的菌液全部移入含 5–10 mL 新鮮 MRS 液體培養(yǎng)基的試管中。
置于 30°C(或 37°C)厭氧或靜置培養(yǎng) 18–24 小時(shí),直至液體呈現(xiàn)明顯的渾濁狀態(tài),證實(shí)增殖成功。
平板劃線與傳代:
取增殖好的菌液,在 MRS 固體瓊脂平板上進(jìn)行分區(qū)分離劃線。
置于厭氧培養(yǎng)箱中培養(yǎng) 24–48 小時(shí),待長出邊緣整齊、表面光滑、呈乳白色或微灰白色的圓形微凸菌落。
挑選單菌落進(jìn)行后續(xù)的實(shí)驗(yàn)或甘油凍存(長期保藏建議添加 20%–30% 無菌甘油置于 -80°C 儲(chǔ)存)。
四、 核心應(yīng)用與科研方向
維生素與生物活性物質(zhì)定量生物測定(Bioassay):該菌株是國際標(biāo)準(zhǔn)的檢測指示菌,廣泛用于微量分析測定:
生物素(Biotin) 測定。
煙酸(Nicotinic acid) 測定。
泛酸(Pantothenic acid) 測定。
對氨基苯甲酸(p-Aminobenzoic acid) 測定。
益生菌微膠囊與腸道遞送系統(tǒng)開發(fā)(Encapsulation):由于其良好的腸道耐受潛力,該菌株頻繁作為標(biāo)準(zhǔn)對象,用于研究海藻酸鈉-蘆薈膠基質(zhì)、油包水/水包油包水(W/O/W)雙重乳液微膠囊包裹技術(shù),以提高益生菌在模擬胃酸(SGF/SGJ)和腸液(SIF)中的存活率。
食品發(fā)酵與保鮮工藝研究:廣泛應(yīng)用于果汁(如刺果番荔枝汁等發(fā)酵)、發(fā)酵乳制品以及青貯飼料的發(fā)酵動(dòng)力學(xué)、代謝產(chǎn)物分析與天然防腐劑(乳酸菌素)的研發(fā)。
PART 2: ENGLISH SECTION
I. General Information and Microbial Background
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 .
II. Morphological and Physiological Properties
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.
III. Recovery and Subculturing Protocols
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.
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