海滨盐薄片菌——分离自海洋盐田的一个嗜盐古菌新种（英文）

【目的】研究分离自我国华南地区阳江盐田的一株嗜盐古菌菌株YJ-41~T,探究其分类学地位。【方法】运用多相分类学方法即通过表型和遗传型特征鉴定,研究菌株YJ-41T的分类学地位。【结果】菌株YJ-41~T的细胞为杆状、革兰氏染色阴性、菌落呈红色。菌株YJ-41T的生长温度范围20–50°C(最适为37°C)、Na Cl浓度范围2.1–4.8 mol/L(最适为3.1 mol/L)、MgCl_2浓度范围0–1.0 mol/L(最适为0.05 mol/L)、pH范围5.0–9.0(最适为pH 7.0)。细胞在蒸馏水中会裂解,维持细胞形态的最低Na Cl浓度为10%(质量体积比)。菌株YJ-41~T的极性脂为磷脂酸(Phosphatidic acid,PA)、磷脂酰甘油(Phosphatidyl glycerol,PG)、磷脂酰甘油磷酸甲基酯(Phosphatidyl glycerol phosphate methyl ester,PGP-Me)、磷脂酰甘油硫酸酯(Phosphatidyl glycerol sulphate,PGS)和8种糖脂;其中3种糖脂为硫酸甘露糖苷葡萄糖二醚(Sulfated mannosyl glucosyl diether,S-DGD-1)、半乳糖苷甘露糖苷葡萄糖二醚(Galactosyl mannosy glucosyl diether,TGD-1)和甘露糖苷葡萄糖二醚(Mannosyl glucosyl diether,DGD-1),其余为未知糖脂。菌株YJ-41~T的16S rRNA基因和rpo B′基因与盐薄片菌属(Halolamina)的成员相关基因相似性分别为97.5%–98.4%和93.1%–94.4%。菌株YJ-41T的G+C mol%为61.4 mol%。【结论】表型、化学分类和系统发育的特性表明,菌株YJ-41T(=CGMCC 1.12859T=JCM 30237T)代表Halolamina属的一个新种,建议命名为海滨盐薄片菌(Halolamina litorea)。     

来源于海洋细菌Altererythrobacter epoxidivorans CGMCC 1.7731T的新型碱性酯酶E22的酶学性质

【目的】克隆表达海洋细菌Altererythrobacter epoxidivorans CGMCC 1.7731~T中的酯酶基因e22,并研究其酶学性质。【方法】分析菌株的全基因组序列,筛选获得一个酯酶基因e22,将其克隆至p ET-28a载体上,并转化至大肠杆菌BL21(DE3)细胞中表达,研究纯化后表达产物的酶学性质。【结果】通过氨基酸序列分析,确定酯酶E22属于脂类水解酶第二家族(Family Ⅱ)。酶学性质研究结果表明,该酶最适反应底物为对硝基苯酚丁酸酯(C4);最适反应pH 10.5,为碱性酯酶;最适反应温度为55°C,并在60°C孵育2 h后仍保留超过50%的活性,显示了良好的热稳定性;1%甲醇、1%Triton X-100或0.1%SDS对酯酶E22的活性无显著影响,而10 mmol/L的Ba~(2+)或Ca~(2+)则对其活性有抑制作用。【结论】E22是一个新型海洋来源酯酶,具有耐碱性、热稳定性、有机溶剂和去垢剂耐受性等优良特性,在工业生产中具有较好的应用潜力。     

一株海洋低温葡萄糖氧化酶菌株的筛选、鉴定及部分酶学性质

【目的】从海洋样品中分离筛选出产葡萄糖氧化酶菌株。【方法】采用双层平板筛选法进行初筛、复筛确定一株酶活较好的菌株,命名为GOD2(Glucose oxidase)。通过形态学、生理生化特征及16S rRNA基因序列分析研究其分类地位,并对其产生的葡萄糖氧化酶进行分离纯化和部分酶学性质的研究。【结果】细菌GOD2为产葡萄糖氧化酶菌株且遗传稳定,初步鉴定该菌株为假单胞杆菌(Pseudomonas migulae),其所产酶最适反应温度为20°C,热稳定性较差,40°C剩余相对酶活80%;超过40°C酶活力迅速下降。【结论】GOD2是一株极具研究价值的产低温葡萄糖氧化酶菌株。目前没有关于利用该菌生产葡萄糖氧化酶的报道。     

一株油藏嗜热厌氧杆菌的分离鉴定和代谢特征的分析

摘要：【目的】从高温油藏中发掘新的微生物种质资源。【方法】采用 Hungate 厌氧操作技术从大港油田采出水中分离到一株厌氧杆菌 HL-3。通过生理生化特征比较和16S rRNA序列比对,确定HL-3的分类地位。【结果】菌株HL-3为严格厌氧的革兰氏阴性杆菌。生长温度范围 40℃－75℃（最适温度 60℃）;pH 范围 5.0－8.0（最适 pH 6.5）;NaCl 浓度范围 0%－3.2%（最适NaCl浓度0.25%）。能够利用葡萄糖、核糖、甘露糖、木糖、纤维二糖等多种碳水化合物,发酵葡萄糖的产物是乙醇、乙酸、CO2及少量丙酸和丁醇。菌株HL-3的（G+C）mol%含量为 33.9%,与Thermoanaerobacter（嗜热厌氧杆菌属）中模式菌株T.uzonensis DSM18761T (EF530067)的16SrRNA 序列相似性为98.8%,与T.sulfurigignens DSM17917T (AF234164)的相似度次之为98.1％。菌株能够耐受浓度较高的亚硫酸根（0.1 mol/L）离子和浓度极高的硫代硫酸根（0.8 mol/L）。当硫代硫酸根浓度高于0.075 mol/L时,菌体内产生硫单质颗粒;同时,在培养血清瓶顶空中检测到硫化氢气体。菌株 HL-3与T.uzonensis DSM18761T对硫代硫酸根和亚硫酸根的耐受程度有很大不同。菌株HL-3对硫代硫酸根和亚硫酸根耐受程度及对硫代硫酸根的代谢机制与T.sulfurigignens DSM17917T(AF234164)极为相似,但二者代谢葡萄糖的产物却极不相同。【结论】所以菌株HL-3可能是Thermoanaerobacter属中的一个新种,其确切分类地位还有待用DNA分子杂交[1]的技术手段做进一步的鉴定。     

红灯食烷菌(Alcanivorax hongdengensis)黄素结合单加氧酶(AlmA)的基因克隆及其烷烃诱导表达

【目的】研究海洋烷烃降解菌新种模式菌株Alcanivorax hongdengensis A-11-3降解长链烷烃的分子机制。【方法】PCR克隆编码黄素结合单加氧酶的基因序列,利用生物信息学软件对序列进行分析,运用RT-PCR和实时荧光定量PCR技术分析基因在不同烷烃诱导下的表达水平。【结果】从菌株A-11-3中克隆获得了两个黄素结合单加氧酶基因片段(almA1和almA2)。它们编码的氨基酸序列与菌株Acinetobacter sp.DSM17874的AlmA同源性分别为58.6%和53.2%。实时荧光定量PCR分析表明,almA1基因只在长链烷烃(C28-C32)的诱导下上调表达,而almA2基因中能在更宽范围的长链烷烃(C24-C34)和支链烷烃诱导下上调表达。两者均在C9-C22的烷烃诱导下没有上调表达。【结论】黄素结合单加氧酶可能是A-11-3降解长链烷烃和支链烷烃的关键酶。     

一株新型同型产乙酸菌Clostridium sp.的自养和异养生长特性

【背景】同型产乙酸菌是一类利用乙酰辅酶A途径固定CO_2合成自身细胞物质并生成乙酸、乙醇等代谢产物的厌氧菌群,其分布广泛、种类繁多且代谢多样。深入研究同型产乙酸菌菌株的代谢能力及特性,对探索该种群的生理生化特性及其环境作用至关重要。【目的】研究一株同型产乙酸菌Clostridium sp. BXX的最适培养条件及其自养与异养生长特性。【方法】设置BXX菌株培养温度10-55°C、初始pH 6.0-9.0、NaCl浓度0-2.0%、不同氮源,测定菌体细胞含量和产物生成浓度,确定菌株最适培养条件。研究BXX菌株分别以H_2/CO_2、合成气、CO、葡萄糖、1,2-丙二醇、甲酸钠、乙二醇甲醚、甘油、丙酮酸和乳酸为底物时的底物消耗、产物生成、菌体细胞含量和pH等,探究其自养和异养生长特性。【结果】BXX菌株的最适培养温度为30°C,初始pH为7.0,NaCl浓度为1.0%,氮源为酵母粉。BXX菌株能以H2/CO2、合成气、葡萄糖、1,2-丙二醇、甲酸钠、乙二醇甲醚和甘油为底物生长,不能以CO、丙酮酸或乳酸为底物生长。【结论】BXX菌株既能自养生长产乙酸,又能异养生长产乙醇。BXX菌株是乙酸发酵的优良菌种资源,有较好的工业应用潜力。     

降解棉秸秆耐热菌株的鉴定及降解条件优化

【目的】鉴定从新疆棉花秸秆高温堆肥中分离出的两株耐热真菌Z1、Z2的属种,并通过优化影响菌株产生纤维素酶的因素来提高菌株对秸秆的降解率。【方法】经形态学和菌株的ITS区克隆与序列分析确定属种,以液体摇瓶发酵产滤纸酶活性(FPA)变化为衡量指标,对Z1、Z2以及二者混合菌(MS)的纤维素酶产生条件进行优化。【结果】菌株Z1为曲霉属烟曲霉(Aspergillus fumigatus Fresen),Z2为蚀丝霉属(Myceliophthora Cost.)。确定Z1以棉秸秆为碳源、以NaNO3为氮源、起始pH 9.5、接种量11%、50°C摇床培养10 d,对棉秸秆降解率为10.19%;Z2以麦秸秆为碳源、以NaNO3为氮源、起始pH 5.5、接种量9%、50°C摇床培养10 d,对麦秆降解率为27.50%;MS以棉花秸秆为碳源、以蛋白胨为氮源、起始pH 5.5、接种量11%、50°C摇床培养10 d,对棉秸秆的降解率为53.45%。【结论】实验表明,MS(Z1、Z2混合)对秸秆的降解效果优于单株菌,降解率达到一半以上,本研究中的两株耐热真菌在降解棉花秸秆、小麦秸秆等农作物废弃秸秆中具有较高的应用价值。     

D-乳酸生产菌菊糖芽孢乳杆菌来源的D-乳酸脱氢酶同工酶

【目的】菊糖芽孢乳杆菌(Sporolactobacillus inulinus)作为典型的同型发酵产D-乳酸的优势菌株,能够高效生产高纯度的D-乳酸。该菌株发酵受到多方面环境因素影响。糖代谢的关键酶例如葡萄糖激酶、磷酸果糖激酶、丙酮酸激酶以及乳酸脱氢酶均为由葡萄糖代谢成为乳酸的关键酶,该菌中相关代谢酶的研究是发酵调控至关重要的基础。分析S.inulinus的基因组表明有3个推测为D-乳酸脱氢酶的基因,其中已有报道研究了1个双功能蛋白[bifunctional protein(BP)]。本研究分别克隆并解析了另2个D-乳酸脱氢酶同工酶的性质。【方法】本研究以S.inulinus Y2-8基因组DNA为模板,克隆得到2个D-ldh基因(dldh、dhdh),经测序分别为D-乳酸脱氢酶[D-lactic acid dehydrogenase(DLDH)]和D-羟基酸脱氢酶[D-isomer specific 2-hydroxyacid dehydrogenase(DHDH)]的基因。构建的重组菌表达蛋白DLDH,DHDH均具有催化丙酮酸生成D-乳酸的功能。【结果】重组菌表达的蛋白经镍柱亲和层析达到电泳纯。SDS-PAGE分析表明DLDH的表观分子量为37 k Da,DHDH的表观分子量为39 k Da。此外,DLDH以丙酮酸为底物时Km值为(0.58±0.04)mmol/L,对底物有较高的亲和力,最适反应温度为35°C,最适p H为6.5;而DHDH以丙酮酸为底物时Km值为(1.70±0.08)mmol/L最适反应温度为30°C,最适p H为7.5。另有报道的BP以丙酮酸为底物时Km值为(3.40±0.02)mmol/L,最适反应温度为30°C,最适p H为5.5。【结论】根据对底物丙酮酸的亲和力,最适温度及最适p H,推测DLDH是乳酸发酵中产D-乳酸的主导催化剂。结合相关酶学性质的分析可为今后的发酵调控提供理论依据。     

产壳聚糖酶菌株的筛选、鉴定及酶学特性分析

【目的】利用筛选培养基,从福建沿海潮间带泥样中分离筛选产壳聚糖酶的菌株,并研究菌株的产酶特性。【方法】通过形态学观察,结合26S rDNA序列进行分类鉴定,采用DNS法测定酶活力。【结果】筛选得到产壳聚糖酶的菌株KQ-1002与草酸青霉(Penicillium oxalicum)的同源性为99%,并初步鉴定为青霉属的一种。发酵培养的最适温度为30°C,最适碳源为1.0%水溶性壳聚糖,最适氮源为1.87%(NH4)2SO4,最适pH为6.0。该菌株液体发酵培养72 h产壳聚糖酶活性最高,经优化后最高产酶量为18 U/mL。纯化后的壳聚糖酶经SDS-PAGE分析其分子量约40 kD。酶促反应最适pH为5.0,最适反应温度为55°C,Km值为1.293 g/L。在离子浓度为1.0×10 3mol/L时,金属离子Cu2+、Hg2+、Ag+对酶的活性均有强烈的抑制作用。壳聚糖酶对不同底物及脱乙酰度的壳聚糖具有不同的降解作用。【结论】筛选获得产壳聚糖酶的真菌菌株KQ-1002的壳聚糖酶活力经优化后提高了约7倍,是一株具有研究和应用潜力的产壳聚糖酶菌株。     

一株苯酚降解菌的鉴定及其降解特性

【目的】鉴定从某化工厂附近土样中分离到的一株耐高浓度苯酚的菌株T10,通过优化菌株的培养条件提高菌株对苯酚的降解率。【方法】根据菌株的形态、生理生化鉴定及16S rDNA测序分析确定其种属,以液体摇瓶培养菌株T10对苯酚的降解率为指标,对菌株的生长条件进行优化。【结果】菌株T10属恶臭假单胞菌(Pseudomonas putida)。添加葡萄糖、蛋白胨能有效缩短T10菌的生长周期,并使苯酚的降解率提高1.7倍。在菌体初始接种浓度为10%、温度为30°C、转速为180 r/min条件下,对初始苯酚浓度、pH和装液量的响应面优化结果如下:初始苯酚浓度3 000 mg/L、pH 7.5和装液量80 mL/250 mL,苯酚去除率最高可达到87.56%。【结论】T10菌能够耐受较高浓度的含酚废水,并且对苯酚有较强的降解能力,为下一步利用生物法处理含酚废水提供科学依据。     

一株嗜热厌氧杆菌的分离、鉴定及其代谢特征

【目的】分离、保护油藏嗜热微生物资源,解析其主要的代谢特征。【方法】利用Hungte厌氧分离技术从大港油田埕海一区油层采出液中分离出厌氧菌株BF1。通过生理生化特征分析、16S rRNA基因序列比对与电化学分析,确定BF1的分类地位及其S元素代谢对腐蚀电流的影响。【结果】菌株BF1为严格嗜热厌氧革兰氏阴性杆菌,顶端产芽孢、不运动,菌体大小为0.42μm×(1.6 5.4)μm,单生、成对或成串生长。其温度生长范围为45°C 75°C(最适温度60°C);pH生长范围在4.5 8.5(最适pH 6.5)之间,比生长速率(μm)0.99 h 1,倍增时间为42 min。能利用葡萄糖、松三糖、棉子糖、甘露糖、乳糖、纤维二糖、果糖、核糖等碳水化合物,利用葡萄糖发酵的产物是乙醇、乙酸、CO2及少量的H2。菌株BF1能还原亚硫酸盐与硫代硫酸盐产生H2S,其耐受上限分别为50 mmol/L和75 mmol/L;还原硫代硫酸钠(50 mmol/L)后其极化电阻由2 099/cm2降低至776/cm2,腐蚀电流由9.936e-006 A提高至3.25e-005 A。细胞膜脂肪酸主要由高级饱和脂肪酸组成,含量最丰富的为十五烷酸占70.6%。菌株BF1的DNA(G+C)mol%含量为34.0%,其16S rRNA与Thermoanaerobacter pseudethanolicus DSM 2355T相似性最高,为98.3%,与T.brockii subsp.brockii DSM 1457T次之,为98.0%。菌株BF1的许多生理、生化特征与T.pseudethanolicus DSM 2355T和T.brockii subsp.brockii DSM 1457T有着明显的差别,如倍增时间、最适生长温度及底物利用等;而菌株BF1的细胞膜脂肪酸组成与T.pseude-thanolicus DSM 2355T也不相同。【结论】菌株BF1可能是Thermoanaerobacter属中的一个新种,其确切分类地位还需要进一步进行DNA分子杂交;其代谢元素硫提高腐蚀电流密度,可能会对油田管道与设备造成腐蚀。     

Paracoccus niistensis sp. nov., isolated from forest soil, India

A Gram-negative, non-motile, catalase-positive and oxidase-positive, aerobic bacterium designated as NII-0918(T) was isolated from soil sample in Western ghat forest, India. 16S rRNA gene sequence analysis showed that strain NII-0918(T) belongs to the subclass α-Proteobacteria, being related to the genus Paracoccus, and sharing highest sequence similarity with Paracoccus chinensis NBRC 104937(T) (99.4%), Paracoccus marinus NBRC 100640(T) (97.3%), Paracoccus koreensis Ch05(T) (97.1%) and Paracoccus kondratievae GB(T) (97.0%). Other members of Paracoccus showed below 97.0% similarity. The DNA-DNA hybridization values between these four strains and NII-0918(T) were 44.7, 28, 32 and 41%, respectively. The major fatty acids of strain NII-0918(T) were summed feature 7 (C18:1 ω7c/ω 9t/ω 12t) (83.0%) and C18:0 (12.5%). Ubiquinone Q-10 was detected as the major respiratory quinone. The G+C content of genomic DNA of NII-0918(T) was 66.6 mol%. On the basis of physiological, morphological, chemotaxonomical and DNA-DNA hybridization data, it is proposed that strain NII-0918(T) should be placed as a novel species, for which we propose Paracoccus niistensis sp. nov. The type strain is NII-0918(T) (CCTCC AA 209055(T) = NCIM 5340(T) = KCTC 22789(T)).     

Paracoccus beibuensis sp. nov., Isolated from the South China Sea

A Gram-negative, non-motile, short rod-shaped or spherical bacterial strain that accumulates poly-β-hydroxybutyrate (PHB) granules was isolated from the Beibu Gulf in the South China Sea. Cells have no polar or subpolar flagella, dividing by binary fission. Phylogenetic analyses based on 16S rRNA gene sequences showed that the strain formed a monophyletic branch at the periphery of the evolutionary radiation occupied by the genus Paracoccus, family Rhodobacteraceae, order Rhodobacterales, class Alphaproteobacteria. The closest neighbours were Paracoccus aestuarii strain B7(T) (97.2% similarity), Paracoccus zeaxanthinifaciens ATCC 21588(T) (97.1% similarity) and Paracoccus homiensis DD-R11(T) (96.8%). The predominant fatty acids were C(18:1) ω7c (82.1%), and significant amounts of C(18:0) (5.6%), C(10:0) 3-OH (2.3%) and C(16:0) (1.5%) were present. The predominant respiratory ubiquinone of strain JLT1284(T) was Q-10 and the DNA G+C content of strain JLT1284(T) was 67.0 mol%. The isolate was also distinguishable from members of the genus Paracoccus on the basis of phenotypic and biochemical characteristics. It is evident from the genotypic, chemotaxonomic and phenotypic data, therefore, that strain JLT1284(T) represents a novel species of the genus Paracoccus, for which the name Paracoccus beibuensis sp. nov. is proposed. The type strain is JLT1284(T)=LMG 24871(T)=CGMCC 1.7295(T)).     

Isolation and characterization of Bacillus thioparus sp. nov., chemolithoautotrophic, thiosulfate-oxidizing bacterium

A novel bacterium, strain BMP-1(T), was isolated from a continuous wastewater treatment culture system operating with a bacterial consortium. Cells of the isolate were Gram-variable, aerobic, moderately halotolerant, motile and endospore-forming rods. Strain BMP-1(T) grew chemolithoautotrophically by oxidation of thiosulfate to sulfate with a growth yield of 1.07 g protein mol(-1) of thiosulfate consumed. DNA G+C content was 43.8 mol%. Its cell wall had peptidoglycan based on m-diaminopimelic acid, and the major component of fatty acid was C(15 : 0). The 16S rRNA gene analysis showed that strain belongs to the genus Bacillus, sharing a 99.5% of sequence similarity with Bacillus jeotgali CCM 7133(T). DNA-DNA hybridization between the isolate of this study and this strain was 44%. Thus, the inclusion of strain BMP-1(T) in the genus Bacillus is suggested as a novel species and the name Bacillus thioparus sp. nov. (Type strain BMP-1(T)=BM-B-436(T)=CECT 7196(T)) is proposed. The sequence of the 16S rRNA gene has been deposited in GenBank with accession number DQ371431.     

Caldicoprobacter algeriensis sp. nov. a New Thermophilic Anaerobic, Xylanolytic Bacterium Isolated from an Algerian Hot Spring

A thermophilic anaerobic bacterium (strain TH7C1(T)) was isolated from the hydrothermal hot spring of Guelma in the northeast of Algeria. Strain TH7C1(T) stained Gram-positive, was a non-motile rod appearing singly, in pairs, or as long chains (0.7-1 × 2-6 μm(2)). Spores were never observed. It grew at temperatures between 55 and 75°C (optimum 65°C) and at pH between 6.2 and 8.3 (optimum 6.9). It did not require NaCl for growth, but tolerated it up to 5 g l(-1). Strain TH7C1(T) is an obligatory heterotroph fermenting sugars including glucose, galactose, lactose, raffinose, fructose, ribose, xylose, arabinose, maltose, mannitol, cellobiose, mannose, melibiose, saccharose, but also xylan, and pyruvate. Fermentation of sugars only occurred in the presence of yeast extract (0.1%). The end-products from glucose fermentation were acetate, lactate, ethanol, CO(2), and H(2). Nitrate, nitrite, thiosulfate, elemental sulfur, sulfate, and sulfite were not used as electron acceptors. The G+C content of the genomic DNA was 44.7 mol% (HPLC techniques). Phylogenetic analysis of the small-subunit ribosomal RNA (rRNA) gene sequence indicated that strain TH7C1(T) was affiliated to Firmicutes, order Clostridiales, family Caldicoprobacteraceae, with Caldicoprobacter oshimai (98.5%) being its closest relative. Based on phenotypic, phylogenetic, and genetic characteristics, strain TH7C1(T) is proposed as a novel species of genus Caldicoprobacter, Caldicoprobacter algeriensis, sp. nov. (strain TH7C1(T) = DSM 22661(T) = JCM 16184(T)).     

Pseudoxanthomonas icgebensis sp. nov., isolated from the midgut of Anopheles stephensi field-collected larvae

A Gram-negative, aerobic, golden yellow, rod-shaped bacterium, a strain designated ICGEB-L15(T), was isolated from the larval midgut of Anopheles stephensi captured in District Jhajjar, Haryana, India. The strain ICGEB-L15(T) grows at 30-50°C (optimum 30-37°C), pH 6.5-8.5 (optimum 7.0-8.0) and in the presence of 2% NaCl. The major fatty acids were iso-C(15:0) (22.5% of total fatty acid), anteiso-C(15:0) (16.5%), iso-C(17:1) 9c (10.3%), iso-C(16:0) (7.3%), C(16:0) (6.1%), and iso-C(11:0) (5.3%). The strain showed the highest 16S rRNA gene sequence similarities with the type strains Pseudoxanthomonas daejeonensis KCTC 12207(T) (97.4%), Pseudoxanthomonas kaohsiungensis J36(T) (97.17%), and Pseudoxanthomonas mexicana AMX 26B(T) (97.11%). The DNA relatedness between ICGEB-L15(T) and Pseudoxanthomonas daejeonensis KCTC 12207(T), Pseudoxanthomonas kaohsiungensis J36(T) and Pseudoxanthomonas mexicana AMX 26B(T) was 24.5%, 28.2%, and 33.6%, respectively. The G+C content of genomic DNA was 69.9 mol%. The major isoprenoid quinone of strain ICGEB-L15(T) was Q-8. The strain ICGEB-L15(T) represents a novel species of the genus Pseudoxanthomonas based on physiological, biochemical and phylogenetic properties; therefore, the name Pseudoxanthomonas icgebensis sp. nov. is proposed. The type strain is ICGEB-L15(T) (=KACC 14090(T) =DSM 22536(T)).     

Corynebacterium aquatimens sp. nov., a lipophilic Corynebacterium isolated from blood cultures of a patient with bacteremia

An unknown lipophilic coryneform bacterium isolated from the blood cultures of a patient with bacteremia was characterized by phenotypic and molecular genetic methods. Chemical analysis revealed the presence of short chain mycolic acids consistent with the genus Corynebacterium. The DNA G+C content was 60.8mol%. Comparative 16S rRNA gene sequence analysis demonstrated that the isolate represents a new subline within the genus Corynebacterium. The closely phylogenetic relative of the unknown bacterium was found to be C. tuscaniense (97.8% sequence similarity). Partial rpoB gene sequence revealed that strain IMMIB L-2475(T) exhibited 13.5% sequence divergence with C. tuscaniense. The unknown bacterium was distinguished from C. tuscaniense by, DNA-DNA hybridization, cellular fatty acid profiles, MALDI-TOF analyses of cell extracts and biochemical tests. Based on the phylogenetic and phenotypic criteria, it is proposed that this bacterium be classified as new species, Corynebacterium aquatimens sp. nov., and is represented by strain IMMIB L-2475(T) (=DSM 45632(T)=CCUG 61574(T)).     

Rhizobium phenanthrenilyticum sp. nov., a novel phenanthrene-degrading bacterium isolated from a petroleum residue treatment system

Strain F11(T), a phenanthrene-degrading bacterium, was isolated from a petroleum residue treatment system, and classified under the genus Rhizobium based on the similarity analysis of its 16S rRNA and recA gene sequences. Strain F11(T) falls into the same phylogenetic clade with Rhizobium oryzae Alt 505(T) (96.8% 16S rRNA gene sequence similarity) and Rhizobium pseudoryzae J34A-127(T) (96.2%). Major cellular fatty acids of strain F11(T) are C(16:0) (6.24%) and summed feature 8 (C(18:1ω7c) and/or C(18:1ω6c), 76.59%), which are also the major fatty acids of R. oryzae Alt 505(T) and R. pseudoryzae J34A-127(T). The DNA G+C content of strain F11(T) was 59.3±0.4 mol%. Based on the phylogenetic analysis as well as biochemical and physiological characteristics, strain F11(T) could be separated from all recognized Rhizobium species. Strain F11(T) (=DSM 21882(T) =CCTCC AB 209029(T)) was considered to be representative of a novel species of Rhizobium, for which the name Rhizobium phenanthrenilyticum sp. nov. is proposed.     

浮霉菌门严格厌氧产氢细菌(Thermopirellula anaerolimosa)的分离及其生理特性

[目的]厌氧颗粒污泥中含有大量未知微生物资源,利用低浓度底物及添加抗生素的培养基进行厌氧发酵细菌的筛选,并对分离菌株进行生理生化特性研究.[方法]利用系列稀释法及亨盖特厌氧滚管技术从制糖废水厌氧处理反应器的颗粒污泥中分离到一株高温厌氧产氢细菌VM20-7T,通过16S rRNA基因序列同源性确定其系统发育地位.[结果]菌株VM20-7T为高温、严格厌氧、革兰氏阴性梨形细菌,细胞大小为(0.7-2.0)μm×(0.7-2.0) μm,不运动,不产芽胞.其生长温度范围为35℃-50℃(最适温度45℃),pH范围为6.0-8.3(最适pH7.0-7.5),NaCl耐受范围为0％-0.5％(w/v,最适浓度0％).菌株VM20-7T可利用葡萄糖、麦芽糖、核糖等多种糖类为唯一碳源生长,葡萄糖发酵终产物是乙酸和H2.该菌株不利用硝酸盐、硫酸盐等作为电子受体生长.G+C含量为60.9 mol％,16S rRNA基因序列同源性显示菌株属于浮霉菌门,但与已培养菌株的同源性较低,与梨形菌属一红小梨形菌属-芽殖小小梨形菌属(Pirellula-Rhodopirellul -Blastopirellula,PRB)分支的亲缘关系最近,但序列相似性也仅为82.7％-84.3％.[结论]利用低浓度糖类并添加抗生素分离厌氧颗粒污泥中的微生物,获得了浮霉菌门首例严格厌氧细菌VM20-7T.生理生化特性和系统发育分析显示,菌株VM20-7T为浮霉菌目的新属新种,命名为Thermopirellula anaerolimosa.该菌株的菌种保藏号为CGMCC 1.5169T=JCM 17478T=DSM 24165T.     

Methylobacterium soli sp. nov. a methanol-utilizing bacterium isolated from the forest soil

A Gram-negative, pink-pigmented, non-spore-forming rod shaped, methanol-utilizing bacterium, strain YIM 48816(T), was isolated from forest soil collected from Sichuan province, China. Strain YIM 48816(T) can grow at 4-37 °C, pH 5.0-7.0 and 0% NaCl (w/v). Based on 16S rRNA gene sequence similarity studies, it belonged to the genus Methylobacterium, and formed a phyletic line. The 16S rRNA gene sequence similarities were 96.2% to Methylobacterium mesophilicum DSM 1708(T) and 96.0% to Methylobacterium brachiatum DSM 19569(T), and the phylogenetic similarities to all other Methylobacterium species with validly published names were less than 96.0%. The major menaquinones detected were Q-10 (97.14%) and Q-9 (2.86%). The major fatty acids were C18:1 ω7c (80.84%). The DNA G + C content was 66.2 mol%. It is apparent from the genotypic and phenotypic data that strain YIM 48816(T) belongs to a novel species of the genus Methylobacterium, for which the name Methylobacterium soli sp. nov. is proposed. The type strain is YIM 48816(T) (CCTCC AA 208027(T) = KCTC 22810(T)).