BioVector® 念珠丝状菌 Kineothrix alysoides DSM 100556 标准菌株

BioVector® Kineothrix alysoides DSM 100556™ / Type Strain

第一部分：中文说明

一、 产品基本信息与分类学背景

• 菌种名称：念珠丝状菌（Kineothrix alysoides）

• 菌株编号：DSM 100556

• 同义菌株别名：KNHs209、ATCC TSD-26

• 分类学地位：厚壁菌门（Bacillota）、梭菌纲（Clostridia）、梭菌目（Eubacteriales）、毛螺菌科（Lachnospiraceae）、丝状菌属（Kineothrix）

• 模式菌株（Type Strain）：是（Yes）。该菌株为 Kineothrix 属的模式建立基准株。

• 物种分离来源：最初由科研团队分离自接种了森林土壤、以柳枝稷（Switchgrass）为底物的厌氧微宇宙演化模型中。

• 生物安全级别：1级（BSL-1）。该菌属于常规环境及共生良性细菌，无已知致病性。

二、 细菌形态学、基因组与生理生化特征

• 细胞与菌落形态：

  • 细胞：革兰氏染色在基因组上归于阳性支系（但具有毛螺菌科典型的易变染色壁结构），呈现细长的杆状，大小约为 $0.5\text{--}0.6\ \mu m \times 3.3\text{--}6.7\ \mu m$。细胞常单个或相连形成极长的念珠状丝状链（Long Filamentous Chains），链长可超过 $30\ \mu m$，在显微镜下可见明显的游动性（具有单生极鞭毛）。能形成抗逆性芽孢。

  • 菌落：在厌氧琼脂平板上培养 48–96 小时后，菌落呈现凸起、圆形、表面光滑、边缘整齐的乳白色或灰白色。

• 基因组特征：

  • GC 含量：42.74 mol%。

  • 核心特征：全基因组测序显示其编码了高达 45 个趋化性受体（Chemotaxis Receptors），这在整个毛螺菌科已知的物种中属于极高水平。

• 代谢特征：

  • 属于严格厌氧菌（Strict Anaerobe）。

  • 具有广泛的糖降解能力（Saccharolytic），能利用多种碳水化合物，但不能利用纤维素（Cellulose）和木聚糖（Xylan）。

  • 主要的碳水化合物发酵产物包括丁酸（Butyrate）、醋酸（Acetate）和甲酸（Formate）。

  • 基因组包含典型的固氮基因，具备在不添加铵盐或尿素的培养基中自主固氮（Nitrogen Fixation）的潜在能力。

三、 培养基配方与理化参数

• 推荐标准培养基：

  • DSMZ Medium 255（经常规预还原脱氧处理的肉汤或琼脂培养基）。

  • 亦可采用标准 PYG（蛋白胨-酵母膏-葡萄糖）培养基进行改良厌氧培养。

• 物理培养参数：

  • 培养温度：30°C 至 37°C（在 30°C 下生长良好，但在 35–40°C 范围内具有最佳生长速率）。

  • 气体环境：100% 严格厌氧（Strictly Anaerobic），操作和孵育均需在厌氧工作站或配有高效氢/氮催化系统的厌氧罐中进行。

  • pH 值：最适生长 pH 为 7.0。

四、 菌株复苏与保存标准步骤

1. 安瓿瓶冻干粉复苏：

   • 将所需的 DSMZ 255 液体培养基置于厌氧环境中进行至少 24 小时的预还原（Pre-reduced），以彻底清除溶解氧。

   • 在厌氧工作台内小心打开安瓿瓶，加入 0.5 mL 预还原的液体培养基将冻干菌块完全重悬。

   • 将悬液转入含有 5–10 mL 新鲜培养基的厌氧试管中，同时可接种至相应的厌氧琼脂平板。

2. 常规孵育：

   • 拧紧管盖，置于 30°C 稳定孵育 48–96 小时，直至液体出现明显的密集成长或浑浊。

3. 长期保存：

   • 采集对数生长末期的菌液，按 1:1 的体积比混合无菌的 30% 甘油保护剂，置于 -80°C 超低温冰箱或液氮中冷冻保存。

五、 核心科研应用方向

1. 肠道微生态与代谢健康研究（脂肪肝与代谢干预）：近年来基础科学研究发现，Kineothrix alysoides 是一种重要的人体与动物肠道共生有益菌。在肥胖或高脂饮食模型中，该菌的丰度会显着下降；作为下一代益生菌（NGPs）候选株，研究显示其能够调节脂质代谢，在缓解代谢功能障碍相关脂肪性肝病（MASLD）及减轻肝脏损伤方面表现出重要的生物学潜力。

2. 高效丁酸生产机制研究：作为新型的毛螺菌科产丁酸指示菌，它常被用于探索肠道内碳水化合物向短链脂肪酸（SCFAs）转化的特定酶促通路。

3. 环境固氮与植物质降解网络构建：由于其独特的分离背景（源自木质纤维素微宇宙）及固氮和高趋化性受体特征，该菌株被广泛应用于解析土壤-植物微生态过渡中的厌氧固氮作用及群落协同代谢网络。

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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