马铃薯干腐病菌硫色镰孢的生物学特性

从碳源、氮源、酸碱度及生长温度等方面对引起马铃薯干腐病的硫色镰孢Fusarium sulphureum的生物学特性进行了研究。结果表明,该病原菌在不同发育阶段对营养和环境条件的要求存在差异。在固体培养基上,菌落生长最佳碳源为葡萄糖、麦芽糖,最佳氮源为蛋白胨,pH8?为最佳;在液体培养基中,菌丝体生长以麦芽糖为最佳碳源,以硝酸钠为最佳氮源,pH6?为最佳;在分生孢子萌发阶段,在以羧甲基纤维素钠、蛋白胨和谷氨酸为碳、氮源的营养液中,分生孢子萌发率最高,最适pH 6–8。该病菌最适生长温度为25℃,分生孢子致死     

Cholesterol is accumulated by mycobacteria but its degradation is limited to non-pathogenic fast-growing mycobacteria

In this report we show that fast-growing non-pathogenic mycobacteria degrade cholesterol from liquid media, and are able to grow on cholesterol as a sole carbon source. In contrast, slow-growing mycobacteria, including pathogenic Mycobacterium tuberculosis and bacillus Calmette-Guérin (BCG), do not degrade and use cholesterol as a carbon source. Nevertheless, pathogenic mycobacteria are able to uptake, modify, and accumulate cholesterol from liquid growth media, and form a zone of clearance around a colony when plated on solid media containing cholesterol. These data suggest that cholesterol may have a role in mycobacterial infection other than its use as carbon source.     

Production and characterization of a new bioemulsifier from Pseudomonas putida ML2

AIMS: To characterize the bioemulsifier produced by a nonfluorescent strain of Pseudomonas putida isolated from a polluted sediment and to determine the influence of pH, temperature, media composition, and carbon and nitrogen source on growth and emulsifying activity. METHODS AND RESULTS: Different indexes were employed to determine the emulsifying properties of culture supernatants of P. putida ML2 in defined and complex media. Surface tension of cell-free supernatants was measured. Purification and chemical analysis of the emulsifier was performed. Confirmed results indicate that a polysaccharide with hexasaccharide repeating units is responsible for the emulsifying activity in a mineral medium with glucose as sole carbon source. Moreover, an emulsifier is produced when growing on naphthalene. CONCLUSIONS: Culture media composition influences the amount and the properties of the emulsifier produced by this P. putida strain. Under nitrogen limiting conditions, a polysaccharide is responsible for the emulsifying activity in defined mineral media. In complex nitrogen rich medium, a different kind of emulsifier is produced. The exopolymer may contribute to hydrocarbons solubilization. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first exopolysaccharide with emulsifying properties produced by a Pseudomonas strain reported to the present. Also chemical composition is significantly different from previous reports. This strain has potential use in bioremediation and the purified polysaccharide may be used in food and cosmetic industry. Moreover, the production of the exopolymer may play a role on biofilm formation.     

Influence of medium composition on the growth and antigen expression of Helicobacter pylori

An evaluation of the ability of various solid and liquid media to support both growth and antigen expression, particularly lipopolysaccharide (LPS) expression, by Helicobacter pylori culture collection strains and clinical isolates was performed. Liquid-based basal media (brain heart infusion, Brucella broth, Mueller–Hinton broth and tryptone soya broth) supported the growth of strains, whereas solid basal media of the same formulation did not support growth. Optimal growth of all strains was obtained on solid and in liquid media containing blood. Supplemented solid media containing supplements other than blood supported growth but only to a small extent. In liquid media excluding blood, serum supplements enhanced growth and horse serum was found to be superior to fetal calf serum. In general, β-cyclodextrin did not increase growth. Mueller–Hinton broth or tryptone soya broth containing horse serum and a nitrogen source such as yeast extract or proteose peptone no. 3 were found to give optimal growth of H. pylori in a blood-free environment. Strains after cultivation in liquid media, irrespective of composition, maintained production of high-molecular weight (mol. wt) LPS with an O side chain independent of medium composition, whereas subculturing on solid media resulted in production of low-mol. wt LPS. Expression of proteins differed in liquid and on solid media, particularly proteins of 57 and 60 kDa, but qualitatively no differences were observed upon supplementation of basal media.     

娄彻氏链霉菌ATCC10739产抗生素Borrelidin发酵条件优化及其分离纯化

对娄彻氏链霉菌ATCC10739固体发酵产生十八元大环内酯抗生素Borrelidin进行了发酵条件的优化。首先筛选得到了理想的发酵培养基;其次考察了发酵时间、起始pH值以及在ISP-2培养基中添加附加碳源、氮源对Borrelidin产量的影响。初步确定最适发酵条件为:ISP-2培养基中添加1%甘油,起始pH值为6.0,培养温度30°C,发酵时间为7 d,产量可达1.336 mg/L。采用有机溶剂萃取、硅胶层析和半制备型高效液相色谱(HPLC)等分离技术纯化得到Borrelidin。     

A chitinolytic actinomycete complex in black soil

A chitinolytic actinomycete complex in chernozem soil has a specific taxonomic composition, which differs from that of the actinomycete complex which is typically isolated on standard nutrient media containing sugars and organic acids as carbon sources. The actinomycete complex that was isolated by using nutrient media with chitin as the source of carbon and nitrogen was dominated by representatives of the genus Streptosporangium, and the actinomycete complex that was isolated by using nutrient media with sugars and organic acids as the carbon sources was dominated by representatives of the genus Streptomyces. The confirmation to the ability of actinomycetes to utilize chitin as a sole source of carbon and nitrogen came from the augmented length and biomass of the mycelium, the increased number and biomass of the actinomycete spores, the production of carbon dioxide, and the accumulation of NH4+ ions in the culture liquid of the actinomycetes that are grown in the nutrient media with chitin.     

Isolation and characterization of Actinopolyspora halophila, gen. et sp. nov., an extremely halophilic actinomycete.

An actinomycete, isolated as a contaminant of a culture medium containing 25% NaCl, has been classified as Actinopolyspora halophila gen. et sp. nov. in the family Nocardiaceae. The morphology and biochemical characteristics of this organism distinguish it from other members of the family Nocardiaceae and other genera possessing a type IV cell wall. It requires high NaCl concentrations for growth and can grow in saturated NaCl. The lowest concentration permitting growth in liquid medium is 12%, and on solid medium, 10%. Colonies developing at lower salt concentrations contain holes resembling viral plaques. No growth occurred in a medium containing 30% KCl instead of NaCl. This organism can grow in simple media with NH4+ salts as nitrogen source and different sugars and other compounds as carbon source. Though it has a salt requirement almost as great as the extremely halophilic rods and cocci, it differs from these in containing diaminopimelic acid and in sensitivity to lysozyme; both properties suggest that it has a mucopeptide cell wall. It also contains some phospholipids common to other actinomycetes, but does not contain any phytanyl ether linked lipids characteristic of other extremely halophilic bacteria.     

A note on a new defined medium for 'Pseudomonas tomato'

A synthetic medium based on d -galactose as a single carbon source and either l -asparagine, l -glutamine or l -threonine as a single nitrogen source has been developed for Pseudomonas tomato. The growth of Ps. tomato on this medium was equal to its growth on commonly used complex media.     

Effect of culture medium composition on the activity of extracellular lectins of Lentinus edodes

The time course of lectin production in culture liquid of the basidial fungus Lentinus edodes strain F-249 in different media under the conditions of submerged culture was studied. The activity of agglutinins depended on the ratio between carbon and nitrogen sources and pH of culture medium. The activity of lectin in culture medium was maximal when the fungus was grown in a medium containing L-arabinose as a source of carbon and L-asparagine as a source of nitrogen (C : N ratio, (9.5-12): 1)) on the day 15-18 of culturing at pH 8-9.     

A Chitinolytic Actinomycete Complex in Chernozem Soil

A chitinolytic actinomycete complex in chernozem soil has a specific taxonomic composition, which differs from that of the actinomycete complex typically isolated on standard nutrient media containing sugars and organic acids as carbon sources. The actinomycete complex that was isolated by using nutrient media with chitin as the source of carbon and nitrogen was dominated by representatives of the genus Streptosporangium, and the actinomycete complex that was isolated by using nutrient media with sugars and organic acids as the carbon sources was dominated by representatives of the genus Streptomyces. The confirmation of the ability of actinomycetes to utilize chitin as a sole source of carbon and nitrogen came from the augmented length and biomass of the mycelium, the increased number and biomass of the actinomycete spores, the production of carbon dioxide, and the accumulation of NH₄ ⁺ ions in the culture liquid of the actinomycetes grown in the nutrient media with chitin.     

Germination of concentrated suspensions of spores from Aspergillus niger

Summary Germination of condiospores fromA.niger in very concentrated suspension was required to inoculate solid fermentation media, but a germination self-inhibitor was detected in spores. It was found that the inhibitory effect depended on the composition of the medium and was reduced when glucose was used as a carbon source. The effect of the self-inhibitor was eliminated by washing the spores and using glucose and a protein nitrogen source in the germination medium. By this method it was possible to increase about 100 times (10⁶ to 10⁸) spore concentration, keeping more than 90% germination.     

碳源、氮源对异养单针藻Monoraphidium sp. FXY-10油脂积累和脂肪酸组成的影响

研究了碳源与氮源对单针藻Monoraphidium sp. FXY-10异养培养的影响。以BG-11为基础培养基,通过添加不同类型、浓度梯度碳源和氮源,比较分析微藻生物量、油脂积累以及脂肪酸组成。结果表明,以葡萄糖作碳源,硝酸钠为氮源,微藻细胞积累的油脂是理想的生物柴油制备原料。硝酸钠浓度分别为1.00、3.00和5.00 g/L时,对油脂产量影响不显著(P>0.05)。葡萄糖浓度为10.00 g/L,硝酸钠为氮源油脂产量达到实验最高值0.84 g/L,其油脂脂肪酸组成主要由C16:0和C18:1等短链饱和脂肪酸和单不饱和脂肪酸组成,不饱和度值(DU)为61.98,相对偏低。     

32P incorporation and growth of the hydrocarbon-degrading pseudomonad UP-2

The marine pseudomonad UP-2 grew, with a doubling time of 1.5 h at 30°C, in media supplemented withn-tetracosane as the sole organic carbon and energy source. Over 99% of the solid hydrocarbon was degraded. In order to overcome serious problems of sampling and measuring growth in the biphase system of hydrocarbon-water, a technique was developed combining the use of separate 1-ml eultures for each sample and³²P labeling as an assay for growth. Using this technique, a growth durve of UP-2 onn-tetracosane was obtained. UP-2 pregrown on sodium acetate can utilize even-chain, liquid, or solidn-alkanes with no observable lag; in contrast, when UP-2 was pregrown onn-tetracosane and transferred to sodium acetate medium, a lag of 1 h was observed.     

Erratum to: Extracellular protein production and morphogenesis of <Emphasis Type="Italic">Lentinula edodes</Emphasis> in submerged culture

Along with a brief review of Lentinula edodes (shiitake mushroom) submerged cultivation history within the framework of important extracellular proteins biosynthesis, this study contains the authors’ own results. The possibility of regulating the lectin activity of shiitake using the synthetic components is shown. The time course of lectin production in culture liquid of L. edodes in different media under submerged culture conditions was studied. The activity of agglutinins depended on the ratio between carbon and nitrogen sources and the pH of the culture medium. A relationship between the chemical composition of nutrient medium, the activity of extracellular lectins of L. edodes, and the formation of pigmented mycelial film in liquid culture has been found. The formulation of medium, on which the brown mycelial film appears in several days of submerged cultivation, is proposed. The results obtained make a contribution to the present notion of biochemical processes that give rise to the occurrence of the aforesaid morphological structure of shiitake. Finally, two extracellular lectins from the submerged culture of L. edodes have been isolated and purified to homogeneity. Their physicochemical properties and composition have been studied.     

工业废水中阿特拉津降解细菌的遗传和生理多样性

【目的】通过遗传学和生理学实验,揭示分离自工业废水的阿特拉津降解细菌具有遗传和生理多样性,为阐明阿特拉津生物降解的分子机理和阿特拉津降解细菌在污染环境生物修复中的应用提供新见解。【方法】用普通PCR方法检测菌株的阿特拉津降解基因,分析其降解基因组成;用基因组重复序列PCR技术(rep-PCR)分析降解菌株的基因组类型;用Western blot方法检测菌株阿特拉津降解途径的第一个酶三嗪水解酶(TrzN);用不同氮源(阿特拉津、莠灭净、扑草净、西玛津、氰草净、阿特拉通和氰尿酸)和碳源(蔗糖、葡萄糖、麦芽糖、乳糖、柠檬酸钠、乙酸钠和琥珀酸钠)培养降解菌株,通过检测培养液的OD600值,证明菌株能够利用的氮源和碳源种类。【结果】对分离自工业废水的27个阿特拉津降解菌株所进行的阿特拉津降解基因PCR检测表明,其降解基因组成分别为trzN-atzBC、trzN-atzABC和atzADEF;通过rep-PCR实验将27个阿特拉津降解菌株分为7个群;Western blot结果表明,27个菌株中有24个含有三嗪水解酶TrzN;氮源利用实验表明,2个菌株能够利用所有7种氮源生长,其余25个菌株只能利用其中的2-6种;碳源利用实验表明,10个菌株能够利用所有7种碳源生长,其余17个菌株只能利用其中的3-6种。【结论】分离自某工业废水的27株阿特拉津降解功能菌存在相当广泛的遗传和生理学上的多样性,trzN-atzABC降解基因组成为首次发现。     

The effect of lactate addition on the growth of Penicillium camembertii on glutamate

The effect of an additional carbon source, lactate, on Penicillium camembertii growth on glutamate as both carbon and nitrogen sources was examined. Glutamate (and lactate) was present in excess in both media. Throughout the whole culture, similar growth time-courses were recorded on both media, indicating the absence of a lactate effect on growth. During the first part of growth, corresponding to an increasing amount of viable biomass, the rate of glutamate consumption remained high, as well as the related ammonium production, indicating its use as a carbon source in addition to being nitrogen source. The low growth rates recorded during the last part of growth resulted in low glutamate consumption, while lactate consumption continued mainly by a maintenance mechanism for the energy supply. A clear differentiation appeared therefore between the carbon source and the energy source: glutamate was mainly used as C source (and N source) for biosynthesis, while lactate was mainly assimilated for energy supply. Carbon and nitrogen yield examinations confirmed this result. Indeed, the C/N ratio found for P. camembertii cellular material (8.14) was about twice that of glutamate (4.29). From this, about half of the available nitrogen was used for biomass formation during growth on glutamate-lactate based medium, as experimentally confirmed (constant yield nitrogen from biomass on nitrogen from glutamate was found (0.49), while the excess nitrogen was released as ammonium). The constant and close to unit (0.99) yield carbon from CO2 on carbon from lactate, also recorded during growth on glutamate-lactate based medium, confirmed that lactate was mainly used as an energy source.     

Nutritional requirements ofLysobacter lactamgenus for the production of cephabacins

Summary To optimize the fermentation medium for the production of new cephem compounds, cephabacins, by an eubacteriaLysobacter lactamgenus IFO 14,288, the effects of medium components on cephabacin production were investigated. Supplementation of glucose as a sole carbon source in liquid media was the best for the antibiotic production as well as for the cell growth. Casamino acid was the best nitrogen source for antibiotic biosynthesis and cell growth among nitrogen sources tested, and this strain could utilize sulfate or thiosulfate as a sulfur source. No significant effects of growth factors (vitamins) on the antibiotic production and cell growth were observed, but ferrous, magnesium and nickel ions slightly enhanced the cephabacin production.     

Comparative studies on the role of nitrogenous compounds in the growth and perithecial development ofChaetomium aureum

Sodium nitrate is a suitable nitrogen source supporting growth and perithecial formation in Chaetomium aureum. 31 inorganic and organic nitrogen sources at a concentration of 0.033% were used at different incubating temperatures (25, 30 and 35 degrees C). The cultures were grown in Petri dishes, culture tubes and Erlenmeyer flasks. An incubating temperature of 30 degrees C was found most suitable for perithecial formation. In general, a nitrogen source in a solid medium could induce better perithecial production than in liquid media.     

Preparation and use of media for protease-producing bacterial strains based on by-products from Cuttlefish (Sepia officinalis) and wastewaters from marine-products processing factories

Cuttlefish powder (CFP) from Sepia officinalis by-products was prepared and tested as a fermentation substrate for microbial growth and protease production by several species of bacteria: Bacillus licheniformis, Bacillus subtilis, Pseudomonas aeruginosa, Bacillus cereus BG1, and Vibrio parahaemolyticus. All microorganisms studied grew well and produced protease activity when cultivated in medium containing only CFP indicating that the strains can obtain their carbon and nitrogen source requirements directly from whole by-product proteins. Moreover, it was found that the addition to the cuttlefish medium of diluted fishery wastewaters (FWW), generated by marine-products processing factories, enhanced the production of protease. Maximum activity was obtained when cells were grown in cuttlefish media containing 5-times or 10-times diluted FWW. Five-times diluted FWW enhanced protease production by B. cereus BG1 and B. subtilis by 467% and 75% more than control media, respectively. The enhancement could have been due to the high organic content or high salts in FWW.As a result, cuttlefish by-products powder enriched with diluted FWW was found to be a suitable growth media for protease-producing strains. This new process, which converts underutilized wastes (liquid and solid) into more marketable and acceptable forms, coupled with protease production, can be an alternative way to the biological treatment of solid and liquid wastes generated by the cuttlefish processing industry.     

根霉液态发酵产生纤溶酶的培养条件优化

目的:目前临床使用的溶栓药物疗效肯定,但还存在许多缺陷,而且价格昂贵,因此研制新型溶栓药物的需求迫切。方法:研究了根霉Rhizopus chinensisYY-15液体摇瓶发酵产生纤溶酶的工艺条件。采用单因素试验对液体发酵培养基的碳源、氮源、碳氮比、初始pH进行了优化;采用正交试验对发酵时间、接种量进行了研究。结果:实验范围内菌株液体发酵产纤溶酶的适宜培养基组成为:麸皮水浓度3%(w/v),豆粕浓度5%(w/v),初始培养基pH5.0。适宜培养条件为接种量6%,培养时间72h。优化条件下的摇瓶液体发酵纤溶酶产量平均达98.31 U/ml。