Effects of fertilizer-based culture media on the production of exocellular polysaccharides and cellular superoxide dismutase by <Emphasis Type="Italic">Phaeodactylum tricornutum</Emphasis> (Bohlin)

Microalgae cultures are receiving attention because of increasing biotechnological and biomedical production of active biomolecules. We evaluated various fertilizer-based culture media to scale up production of the marine microalga Phaeodactylum tricornutum for production of exocellular polysaccharides (EPS), soluble proteins, and cellular superoxide dismutase (SOD). The standard source of sodium nitrate was the same as that used in the synthetic f/2 culture medium and ammonium nitrate, urea, ammonium sulfate, and calcium nitrate as alternative sources of nitrogen. The maximum production of EPS was achieved in microalgae cells grown in the culture media containing 63 and 23% nitrogen from ammonium sulfate, and also in microalgae cells grown in the culture media containing 3% nitrogen from ammonium nitrate. The maximum production of cellular SOD was achieved in microalgae cells grown in the culture media containing 35 and 26% nitrogen from ammonium sulfate, and in the culture media containing 17% nitrogen from urea. The results suggest that it is possible to use a source of nitrogen, other than sodium nitrate, to scale up growth of P. tricornutum for production of EPS and SOD at reduced costs.     

铁尾矿区油松根际溶磷泛菌D2的筛选鉴定及溶磷特性

从河北省迁安市马兰庄镇铁尾矿植被恢复区油松根际分离出2株溶磷细菌,经过平板初筛和摇瓶复筛得到1株溶磷能力较强的菌株D2.通过菌落形态、生理生化特性及16S rDNA序列分析,确定此菌株D2属于泛菌属.利用液体发酵试验测定不同碳源、氮源对菌株D2溶磷能力的影响,通过高效液相色谱测定D2在不同氮源条件下产生有机酸的种类和浓度.结果表明:菌株D2对磷酸三钙有较强的溶磷能力,培养液有效磷含量最高为392.13 mg·L^-1,菌株D2的溶磷能力在碳源为葡萄糖、氮源为硫酸铵时效果最好;高效液相色谱测定发现,不同氮源条件下,D2分泌有机酸的种类和浓度存在差异,以硫酸铵、氯化铵、硝酸钾、硝酸钠、硝酸铵为氮源,均产生草酸、甲酸、乙酸和柠檬酸,以硫酸铵、氯化铵、硝酸铵为氮源还产生苹果酸.相关性分析表明,乙酸含量与有效磷含量间呈显著正相关(r=0.886,P<0.05),表明溶磷泛菌D2分泌的乙酸对无机磷的溶解有明显的促进作用,这也很可能是该菌株的重要溶磷机制之一.     

基质氮素对大球盖菇生长及亚硝酸盐含量的影响

目的探索氮源对大球盖菇生长及亚硝酸盐、硝酸盐含量的影响。方法在培养基中添加不同含氮化合物,培养菌丝,定时测定生长量,并采用重氮偶合分光光度法测定其亚硝酸盐、硝酸盐含量。结果添加亚硝酸盐的基质,菌丝生长速度比对照慢。尿素、硫酸铵、硝酸铵和硝酸钾四处理之间差异无显著性,其生长速度最快。各处理间亚硝酸盐、硝酸盐含量差异均存在非常显著性。亚硝酸钾处理,亚硝酸盐含量高达402.03 mg/kg,不宜食用。硝酸钾处理,亚硝酸盐含量为30.87 mg/kg,食用100 g,就会超出亚硝酸盐日限量;硝酸铵处理,硝酸盐含量远远超过日常蔬菜,也不宜食用。结论在栽培大球盖菇时氮源不宜使用亚硝酸盐、硝酸盐,应使用尿素或硫酸铵,有机氮源也可以。     

Effects of the addition of different nitrogen sources in the tequila fermentation process at high sugar concentration

AIMS: To study the effect of the addition of different nitrogen sources at high sugar concentration in the tequila fermentation process. METHODS AND RESULTS: Fermentations were performed at high sugar concentration (170 g l(-1)) using Agave tequilana Weber blue variety with and without added nitrogen from different sources (ammonium sulfate; glutamic acid; a mixture of ammonium sulfate and amino acids) during the exponential phase of growth. All the additions increased the fermentation rate and alcohol efficiency. The level of synthesis of volatile compounds depended on the source added. The concentration of amyl alcohols and isobutanol were decreased while propanol and acetaldehyde concentration increased. CONCLUSIONS: The most efficient nitrogen sources for fermentation rate were ammonium sulfate and the mixture of ammonium sulfate and amino acids. The level of volatile compounds produced depended upon types of nitrogen. The synthesis of some volatile compounds increased while others decreased with nitrogen addition. SIGNIFICANCE AND IMPACT OF THE STUDY: The addition of nitrogen could be a strategy for improving the fermentation rate and efficiency in the tequila fermentation process at high sugar Agave tequilana concentration. Furthermore, the sensory quality of the final product may change because the synthesis of the volatile compounds is modified.     

氮源流加对Alcaligenes eutrophus 积累聚β-羟基丁酸的影响

以真养产碱杆菌（Ａｌｃａｌｉｇｅｎｅｓ ｅｕｔｒｏｐｈｕｓ）为聚β－羟基丁酸９ＰＨＢ）的生产菌株,在分析了ＰＨＢ发酵过程参数变化的基础上,进一步探讨了ＰＨＢ合成期不同的硫酸铵流加速度对ＰＨＢ合成的影响。研究结果表明,在ＰＨＢ合成阶段,培养基中氮源的完全缺乏。导致细胞合成ＰＨＢ能力的下隆;在ＰＨＢ合成期,不同的氮源流加速率对ＰＨＢ合成过程存在着显著的影响,当流加速率较小时,尽管最终胞内ＰＨＢ含量很高     

Effect of nitrogen sources on cellulase biosynthesis by a mutant strain of Trichoderma viride 44

The effect of various nitrogen sources on cellulase biosynthesis by the mutant strain Trichoderma viride 44 was examined. This strain may utilized nitrogen in the nitrate, ammonium of organic form. When cultivating this strain, it appears advantageous to add to the nutrient medium yeast and yeast lyzates as well as their mixture with ammonium sulfate. Cellulase reached its maximum activity of 20.2, 21.5 and 23.2 mu/ml when grown on the medium containing ammonium phosphate, peptone and brewing yeast plus ammonium sulfate, respectively. It is useful to apply nitrogen in its organic forms in small quantities and in combination with mineral forms. The nitrogen presence in the medium is necessary only at the exponential stage of fungal growth. The lack of nitrogen in the stationary stage characterized by the maximum cellulase formation does not inhibit an increase in the enzyme activity.     

Medium optimization of carbon and nitrogen sources for the production of spores from Bacillus amyloliquefaciens B128 using response surface methodology

The production of spores from Bacillus amyloliquefaciens B128 was investigated in shaker-flask cultivation. Optimization of the culture medium was carried out by using a two-step approach. A quick identification of a suitable source of carbon and nitrogen was obtained by a simple screening experiment, which was followed by an application of response surface methodology (RSM) for further optimization design. A five-level four-factor central composite design was employed to determine the maximum spore yield at optimum levels for lactose, tapioca, ammonium sulfate and peptone. Tapioca and peptone showed a significant linear main effect, while lactose and ammonium sulfate had no significant linear effect. The spore production was also significantly affected by lactose–ammonium sulfate and lactose–peptone. Optimum cultivation parameters were (g/L): 12.7 of lactose, 16.7 of tapioca, 1.8 of ammonium sulfate and 8.0 of peptone. The prediction spore yield was 5.93 × 10⁸ (no/mL). The actual experimental results were in agreement with the prediction.     

绿色木霉JFY-14产纤维素酶的产酶条件研究

利用实验室现有的纤维素酶高产菌株制备纤维素酶。考察了JFY-14菌株产酶培养过程中pH值、培养时间、氮源等条件的影响。得到了最佳产纤维素酶条件:培养时间为72~75 h,初始pH值为4.5~5.0以及最佳培养氮源为1%的硫酸铵。     

Oscillatory growth rate responses of S. Cerevisiae in continuous culture

The yeast S. cerevisiae was grown on dilute, chemically defined media in continuous culture with either glucose or ammonium sulfate as the growth-limiting ingredient. Changes in dilution rate or glucose concentration induced decaying oscillations in the numbers of yeast growing on ammonium sulfate-limited media. Spot checks indicated that Cell dry weight and Kjeldahl nitrogen followed the cell numbers during these oscillations. With glucose-limited media, there was no response to step changes in ammonium sulfate concentration, and dilution rate step changes gave non-oscillatory transient responses.     

产多聚唾液酸的菌种筛选及产酸条件

通过对40株大肠杆菌进行产多聚唾液酸的筛选,得到一株高产多聚唾液酸菌株C-8,对该菌的一系列培养条件进行了研究。最佳培养基为：山梨醇2.5％,硫酸铵0.5％,磷酸氢二钾90mmol／L,胰蛋白陈1.5％,硫酸镁0.04％,pH7．8。在37℃,250r／min摇床培养65h,可使菌体在每毫升培养液中产多聚唾液酸1200μg。     

Ureidosuccinic Acid Uptake in Yeast and Some Aspects of Its Regulation

Ureidosuccinic acid (USA) is an intermediary product in pyrimidine biosynthesis. When proline was the sole nitrogen source, USA uptake occurred; however, when ammonium sulfate or glutamic acid was the nitrogen source, uptake was inhibited. Thus, a ura2 strain which does not synthesize USA would not grow when this substance was supplied on an ammonium sulfate or glutamic acid medium. Mutants are described in which uptake was constitutive on such a medium. Permeaseless mutants for USA have been found, and evidence is presented for permease specificity. It is shown that all constitutive mutants use the same transport system that is missing in the permeaseless mutant. These mutants are constitutive for two permeases: the specific USA permease and the general amino acid permease. The transport system studied here, like the general amino acid transport system, is regulated by nitrogen metabolism. These facts and others suggest that our permease constitutive mutants are impaired in nitrogen metabolism.     

Inorganic nitrogen source for autotrophic growth of Euglena mutabilis Schmitz

The effect of inorganic nitrogen source on population growth of Euglena mutabilis, an acidophillic benthic protozoa colonizing on the sediment of acid mine drainage, was investigated. Sodium nitrate, ammonium chloride, and ammonium sulfate were tested as nitrogen sources. The population density of E. mutabilis at equilibrium density cultivated in ammonium chloride‐ and ammonium sulfate‐containing media was 9–11 times higher than that in sodium nitrate‐containing medium at the optimal salt medium concentration. The population growth of E. mutabilis in ammonium sulfate‐containing medium was rapid and reached half of the equilibrium density after ca. 228 h, which was ca. 77 h earlier than that in ammonium chloride‐containing medium. Culture medium with ammonium sulfate as the nitrogen source achieved the highest maximum population density and the fastest growth rate among the three nitrogen salts used as nitrogen sources.     

白葡萄酒活性干酵母对不同氮源利用的研究

选用5种不同的白葡萄酒活性干酵母,以硫酸铵、氯化铵、硝酸铵、尿素、酵母粉等5种物质为氮源,观察其生长量并称量菌体重,以此分析其对氮源利用情况及不同氮源对酵母生长的影响。研究表明:供试菌系在不同氮源中均能生长。不同氮源对酵母的生长速度和生长量有不同影响;不同酵母菌种对不同氮源的利用也有差异。在以酵母粉为氮源的培养基中生长最好。在实验提供的氮源中,酵母粉为供试菌最优氮源,其次是硫酸铵,氯化铵与硫酸铵基本相当,而硝酸铵最差。8#菌种对各种氮源的利用能力相对较强。17#菌种对各种氮源的利用能力最弱。     

A method for developing a controlled process. Development and execution of a program supplying culture medium components for rifamycin B biosynthesis

The study provided data on the rate of consumption of the main nutrients by the rifamycin-producing culture in relation to their content in the initial medium. A program for feeding ammonium sulfate and cornsteep liquor and joint control of both the nutrients was developed and tested. The results of the testing confirmed the practicable precision of the feeding. The control of cornsteep liquor and ammonium sulfate concentrations with the program allowed to increase the yield of rifamycin B by 15 and 20%, respectively. The joint control increased the level of the required product by 25%.     

NUTRITIONAL REQUIREMENTS FOR OVULE FORMATION IN EXCISED PISTILS OF NIGELLA

The nutritional requirements for ovule formation in Nigella saliva L. were investigated by growing excised pistils on defined media. Pistils grown on a medium containing the minerals of Murashige and Skoog produced significantly more ovules than on a medium containing the minerals of Bilderback. When the nitrogen, sulfate, and phosphate of the Bilderback medium were adjusted to levels comparable to those of the Murashige and Skoog medium, a similar number of ovules was formed. The effect of different forms and concentrations of nitrogen on ovule formation and pistil growth was investigated. High concentrations of nitrate (40 mil) favored pistil growth and ovule formation, but comparable levels of ammonium were toxic. When ammonium at concentrations above 10 mM was added to nitrate media, ovule formation was inhibited. A medium containing low concentrations of ammonium (10 mM) and nitrate (5 mM) supported more ovule formation and pistil growth in young pistils than a low-nitrate (5 mM) medium without ammonium. However, ovule formation on a medium containing 10 mM ammonium and 5 mM nitrate was significantly less than on a medium containing only 15 mM nitrate. Low concentrations of organic nitrogen in the form of α-alanine (1 mM) and γ-aminobutyric acid (5 mM) supported ovule formation and pistil growth similar to a high nitrate medium.     

Regulation of nitrogen catabolic enzymes in Streptomyces clavuligerus

The levels of several enzymes involved in assimilation of different nitrogen compounds were investigated in Streptomyces clavuligerus in relation to the nitrogen source supplied to the cultures. Threonine dehydratase, serine dehydratase, proline dehydrogenase, histidase and urocanase were not decreased in the presence of ammonium. The latter two enzymes were induced by histidine in the culture medium, while proline dehydrogenase was induced by proline. Glutamine synthetase, urease and ornithine aminotransferase levels were higher with poor nitrogen sources and were repressed by ammonium. Arginase was induced by arginine and repressed by ammonium. Glutamine synthetase was rapidly inactivated upon addition of ammonium to the culture, and could be reactivated in vitro by treatment with snake venom phosphodiesterase, which suggested that adenylylation is involved in the inactivation. Three previously isolated mutants with abnormal glutamine synthetase activities showed pleiotropic effects on urease formation. All these data point to a mechanism controlling preferential utilization of some nitrogen sources in this species.     

Effect of nitrogen source on biosynthesis of rapamycin by Streptomyces hygroscopicus

Six non-amino acid nitrogen compounds were examined as nitrogen source for growth of Streptomyces hygroscopicus and biosynthesis of rapamycin. Of the nitrogen sources studied, ammonium sulfate was the best with respect to formation of rapamycin, and supported cell growth comparable to the organic nitrogen sources used in the control chemically defined medium, ie, aspartate, arginine plus histidine. In the new chemically defined medium, which is buffered with 200 mM 2-(N-morpholino)ethanesulfonic acid to prevent decline of pH during fermentation, an ammonium sulfate concentration of 40 mM was optimal for biosynthesis of rapamycin. Rapamycin production increased by more than 30% on both volumetric and specific bases as compared to the previous medium containing the three amino acids as nitrogen source. Received 08 November 1996/ Accepted in revised form 07 April 1997     

L-Glutamate oxidase from Streptomyces cremeus 510 MGU: effect of nitrogen sources on enzyme secretion]

Effect of nitrogen sources (organic complexes and mineral salts) on L-glutamate oxidase synthesis by Streptomyces cremeus 510 MGU was studied. Optimal enzyme production was not provided by any single nitrogen source. The most effective combination of nitrogen sources (soy flour, peptone, ammonium sulfate) was elaborated by the mathematical planning method. The results of experiment allowed to enhance biosynthesis of extracellular L-glutamate oxidase to 2.4-2.6 U/mL. It was shown that L-glutamate oxidase of Streptomyces cremeus 510 MGU is highly specific to substance and is stable during storage of filtrated culture with pH 6.8-9.0.     

Effect of the addition of nitrogen sources to cassava fiber and carbon-to-nitrogen ratios on Agaricus brasiliensis growth

The same substratum formulation to grow Agaricus bisporus has been used to grow Agaricus brasiliensis since its culture started in Brazil. Despite being different species, many of the same rules have been used for composting or axenic cultivation when it comes to nitrogen content and source in the substrate. The aim of this study was to verify the mycelial growth of A. brasiliensis in different ammonium sulfate and (or) urea concentrations added to cassava fiber and different carbon-to-nitrogen (C:N) ratios to increase the efficiency of axenic cultivation. Two nitrogen sources (urea and (or) ammonium sulfate) added to cassava fiber were tested for the in vitro mycelial growth in different C:N ratios (ranging from 2.5:l to 50:l) in the dark at 28 degrees C. The radial mycelial growth was measured after 8 days of growth and recorded photographically at the end of the experiment. Nitrogen from urea enhanced fungal growth better than ammonium sulfate or any mixture of nitrogen. The best C:N ratios for fungal growth were from 10:l to 50:l; C:N ratios below 10:l inhibited fungal growth.     

Control of dimorphism in a biochemical variant of Candida albicans

The cellular morphology of a biochemical variant of Candida albicans could be controlled by the ratio of carbon dioxide to oxygen in the culture system or by individual amino acids. Predominantly pseudohyphal morphology was observed (i) at a CO(2) to O(2) ratio of 2:1 and (ii) without the addition of carbon dioxide, when either glycine, d- or l-ornithine, l-serine, l-methionine, l-phenylalanine, or l-tyrosine was the sole nitrogen source in the culture medium. When ammonium chloride, ammonium sulfate, l-glutamic acid, l-glutamine, or l-proline was the nitrogen source, yeastlike growth was observed in the presence or absence of CO(2). More adenosylmethionine was present in pseudohyphal than in yeastlike cells, and pseudohyphal cell wall preparations contained less methionine than cell walls from the yeastlike form. These results suggest a correlation between sulfur amino acid metabolism and dimorphism.