BioVector? 念珠絲狀菌 Kineothrix alysoides DSM 100556 標(biāo)準(zhǔn)菌株

BioVector? Kineothrix alysoides DSM 100556? / Type Strain

第一部分：中文說(shuō)明

一、 產(chǎn)品基本信息與分類學(xué)背景

• 菌種名稱：念珠絲狀菌（Kineothrix alysoides）

• 菌株編號(hào)：DSM 100556

• 同義菌株別名：KNHs209、ATCC TSD-26

• 分類學(xué)地位：厚壁菌門(mén)（Bacillota）、梭菌綱（Clostridia）、梭菌目（Eubacteriales）、毛螺菌科（Lachnospiraceae）、絲狀菌屬（Kineothrix）

• 模式菌株（Type Strain）：是（Yes）。該菌株為 Kineothrix 屬的模式建立基準(zhǔn)株。

• 物種分離來(lái)源：最初由科研團(tuán)隊(duì)分離自接種了森林土壤、以柳枝稷（Switchgrass）為底物的厭氧微宇宙演化模型中。

• 生物安全級(jí)別：1級(jí)（BSL-1）。該菌屬于常規(guī)環(huán)境及共生良性細(xì)菌，無(wú)已知致病性。

二、 細(xì)菌形態(tài)學(xué)、基因組與生理生化特征

• 細(xì)胞與菌落形態(tài)：

  • 細(xì)胞：革蘭氏染色在基因組上歸于陽(yáng)性支系（但具有毛螺菌科典型的易變?nèi)旧诮Y(jié)構(gòu)），呈現(xiàn)細(xì)長(zhǎng)的桿狀，大小約為 $0.5\text{--}0.6\ \mu m \times 3.3\text{--}6.7\ \mu m$。細(xì)胞常單個(gè)或相連形成極長(zhǎng)的念珠狀絲狀鏈（Long Filamentous Chains），鏈長(zhǎng)可超過(guò) $30\ \mu m$，在顯微鏡下可見(jiàn)明顯的游動(dòng)性（具有單生極鞭毛）。能形成抗逆性芽孢。

  • 菌落：在厭氧瓊脂平板上培養(yǎng) 48–96 小時(shí)后，菌落呈現(xiàn)凸起、圓形、表面光滑、邊緣整齊的乳白色或灰白色。

• 基因組特征：

  • GC 含量：42.74 mol%。

  • 核心特征：全基因組測(cè)序顯示其編碼了高達(dá) 45 個(gè)趨化性受體（Chemotaxis Receptors），這在整個(gè)毛螺菌科已知的物種中屬于極高水平。

• 代謝特征：

  • 屬于嚴(yán)格厭氧菌（Strict Anaerobe）。

  • 具有廣泛的糖降解能力（Saccharolytic），能利用多種碳水化合物，但不能利用纖維素（Cellulose）和木聚糖（Xylan）。

  • 主要的碳水化合物發(fā)酵產(chǎn)物包括丁酸（Butyrate）、醋酸（Acetate）和甲酸（Formate）。

  • 基因組包含典型的固氮基因，具備在不添加銨鹽或尿素的培養(yǎng)基中自主固氮（Nitrogen Fixation）的潛在能力。

三、 培養(yǎng)基配方與理化參數(shù)

• 推薦標(biāo)準(zhǔn)培養(yǎng)基：

  • DSMZ Medium 255（經(jīng)常規(guī)預(yù)還原脫氧處理的肉湯或瓊脂培養(yǎng)基）。

  • 亦可采用標(biāo)準(zhǔn) PYG（蛋白胨-酵母膏-葡萄糖）培養(yǎng)基進(jìn)行改良厭氧培養(yǎng)。

• 物理培養(yǎng)參數(shù)：

  • 培養(yǎng)溫度：30°C 至 37°C（在 30°C 下生長(zhǎng)良好，但在 35–40°C 范圍內(nèi)具有最佳生長(zhǎng)速率）。

  • 氣體環(huán)境：100% 嚴(yán)格厭氧（Strictly Anaerobic），操作和孵育均需在厭氧工作站或配有高效氫/氮催化系統(tǒng)的厭氧罐中進(jìn)行。

  • pH 值：最適生長(zhǎng) pH 為 7.0。

四、 菌株復(fù)蘇與保存標(biāo)準(zhǔn)步驟

1. 安瓿瓶?jī)龈煞蹚?fù)蘇：

   • 將所需的 DSMZ 255 液體培養(yǎng)基置于厭氧環(huán)境中進(jìn)行至少 24 小時(shí)的預(yù)還原（Pre-reduced），以徹底清除溶解氧。

   • 在厭氧工作臺(tái)內(nèi)小心打開(kāi)安瓿瓶，加入 0.5 mL 預(yù)還原的液體培養(yǎng)基將凍干菌塊完全重懸。

   • 將懸液轉(zhuǎn)入含有 5–10 mL 新鮮培養(yǎng)基的厭氧試管中，同時(shí)可接種至相應(yīng)的厭氧瓊脂平板。

2. 常規(guī)孵育：

   • 擰緊管蓋，置于 30°C 穩(wěn)定孵育 48–96 小時(shí)，直至液體出現(xiàn)明顯的密集成長(zhǎng)或渾濁。

3. 長(zhǎng)期保存：

   • 采集對(duì)數(shù)生長(zhǎng)末期的菌液，按 1:1 的體積比混合無(wú)菌的 30% 甘油保護(hù)劑，置于 -80°C 超低溫冰箱或液氮中冷凍保存。

五、 核心科研應(yīng)用方向

1. 腸道微生態(tài)與代謝健康研究（脂肪肝與代謝干預(yù)）：近年來(lái)基礎(chǔ)科學(xué)研究發(fā)現(xiàn)，Kineothrix alysoides 是一種重要的人體與動(dòng)物腸道共生有益菌。在肥胖或高脂飲食模型中，該菌的豐度會(huì)顯著下降；作為下一代益生菌（NGPs）候選株，研究顯示其能夠調(diào)節(jié)脂質(zhì)代謝，在緩解代謝功能障礙相關(guān)脂肪性肝病（MASLD）及減輕肝臟損傷方面表現(xiàn)出重要的生物學(xué)潛力。

2. 高效丁酸生產(chǎn)機(jī)制研究：作為新型的毛螺菌科產(chǎn)丁酸指示菌，它常被用于探索腸道內(nèi)碳水化合物向短鏈脂肪酸（SCFAs）轉(zhuǎn)化的特定酶促通路。

3. 環(huán)境固氮與植物質(zhì)降解網(wǎng)絡(luò)構(gòu)建：由于其獨(dú)特的分離背景（源自木質(zhì)纖維素微宇宙）及固氮和高趨化性受體特征，該菌株被廣泛應(yīng)用于解析土壤-植物微生態(tài)過(guò)渡中的厭氧固氮作用及群落協(xié)同代謝網(wǎng)絡(luò)。

PART 2: ENGLISH SECTION

I. General Information and Taxonomic Background

• Organism Name: Kineothrix alysoides

• Strain Identification: DSM 100556 (Type Strain)

• Alternative Strain Aliases: KNHs209, ATCC TSD-26

• Taxonomic Lineage: Phylum Bacillota; Class Clostridia; Order Eubacteriales; Family Lachnospiraceae; Genus Kineothrix.

• Type Strain Designation: Yes. Serves as the official standard nomenclatural type strain for the genus Kineothrix.

• Isolation Source: Originally isolated from a switchgrass-fueled anaerobic microcosm seeded with forest soil in Greenfield, Massachusetts, USA.

• Biosafety Level: BSL-1. Commensal anaerobic bacterium lacking known virulence factors or pathogenic potential toward humans.

II. Morphological, Genomic, and Physiological Attributes

• Cell & Colony Morphology:

  • Cells: Phylogenetically Gram-positive but possessing the thin cell-wall structure characteristic of the family Lachnospiraceae. Appears as slender rods measuring $0.5\text{--}0.6\ \mu m \times 3.3\text{--}6.7\ \mu m$. Cells regularly arrange into distinctive, long filamentous chains (resembling a necklace or string of beads) exceeding $30\ \mu m$ in length. Highly motile via a single polar flagellum and capable of forming endospores.

  • Colonies: On anaerobic agar, colonies developing between 48 and 96 hours are convex, circular, glossy, off-white, and possess entire margins.

• Genomic Characteristics:

  • DNA G+C Content: 42.74 mol%.

  • Key Genomic Feature: Encodes an exceptionally high number of chemotaxis receptors (45 genes), marking one of the highest genomic sensor counts recorded within the family Lachnospiraceae.

• Metabolic Pattern:

  • Classed as a strict (obligate) anaerobe.

  • Broadly saccharolytic, efficiently utilizing various carbohydrates, but unable to degrade cellulose or xylan.

  • Ferments carbohydrates to yield butyrate, acetate, and formate as its principal volatile fatty acid end-products.

  • Possesses functional nitrogen fixation genes, showing the metabolic potential for independent nitrogen ($N_2$) fixation in ammonium-depleted habitats.

III. Medium Composition and Physical Cultivation Metrics

• Recommended Cultivation Media:

  • DSMZ Medium 255 (An anaerobic, pre-reduced formulation suitable for Lachnospiraceae maintenance).

  • Alternatively, pre-reduced Peptone-Yeast-Glucose (PYG) broth supplemented with specific saccharide streams under sterile conditions.

• Physical Environmental Windows:

  • Incubation Temperature: Routinely cultured at 30°C to 37°C (The repository recommends 30°C, though optimal biological metabolic growth peaks between 35°C and 40°C).

  • Gaseous Headspace: 100% Strictly Anaerobic atmosphere. All manipulation, handling, and growth cycles must exclude oxygen.

  • pH Threshold: Optimally maintained at a neutral pH of 7.0.

IV. Recovery and Cryopreservation Protocols

1. Rehydration from Freeze-Dried Pellets:

   • Utilize strictly pre-reduced, deoxygenated DSMZ Medium 255 liquid broth inside an anaerobic workstation.

   • Crack the outer glass ampoule safely, introduce 0.5 mL of the oxygen-free medium to totally resuspend the dried pellet, and transfer the mixture back into a primary anaerobic tube (5–10 mL).

   • Streak a small loopful onto an anaerobic agar plate for purity confirmation.

2. Incubation Phase:

   • Seal tightly and incubate at 30°C for 48–96 hours until visible cloudiness or deep turbidity signals active biomass exponential expansion.

3. Long-Term Cryopreservation:

   • Harvest active late-log phase cultures. Mix thoroughly with an equal volume of a sterile anaerobic 30% glycerol solution to reach a 15% final preservative balance, and store immediately in an ultra-low freezer (-80°C) or a liquid nitrogen depot.

V. Strategic Research Verticals

1. Next-Generation Probiotics (NGPs) & Metabolic Health (MASLD Mitigation): Emerging biomedicine models demonstrate that Kineothrix alysoides acts as a crucial gut commensal that drops in abundance under high-fat dietary conditions. Oral administration of this strain significantly mitigates metabolic dysfunction-associated steatotic liver disease (MASLD) by correcting host lipid accumulation, restoring gut-liver axis balance, and lowering liver injury indices.

2. Intestinal Butyrate Production Kinetics: Serves as a prime type model within the Lachnospiraceae family to evaluate short-chain fatty acid (SCFA) conversion pathways, supporting gut epithelial cell barrier reinforcement and anti-inflammatory research.

3. Anaerobic Nitrogen Fixation & Environmental Biomass Grids: Investigated for its specialized soil-microcosm-derived traits to track how high chemotactic sensing profiles and non-symbiotic nitrogen fixation dynamics govern microbial network survival in oxygen-depleted terrestrial biomes.

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