Accumulation and antioxidant activity of secondary carotenoids in the aerial microalga Coelastrella striolata var. multistriata

The growth and nitrate uptake of the aerial microalga Coelastrella striolata var. multistriata, which was isolated from the surface of rocks, were characterized under a variety of conditions in this study. The maximum specific growth rate of the alga, having prominent inorganic nitrogen uptake in the fresh medium, was 0.30 d⁻¹, as calculated in the growth logarithmic phase. It was also shown that the alga had abilities to be reddish orange to green colour (depending on the nitrogen source concentration in the medium) and to synthesize very high amounts of a complex mixture of carotenoids, such as canthaxanthin, astaxanthin and β-carotene. The reddish orange cells of the alga could accumulate 56.0 mg of major secondary carotenoids per g biomass. In the content of carotenoids, canthaxanthin, astaxanthin and β-carotene in the cells were 47.5, 1.5, and 7.0 mg/g dwc, respectively. Additionally, it was shown that the algal extract containing those carotenoids had an antioxidant potential in lipid foods. In the near future, the aerial microalga C. striolata var. multistriata will be used as a functional material in a variety of commercial applications, such as feed supplements, natural antioxidants and food dyes.     

碳酸氢铵为氮源对螺旋藻培养的影响

以碳酸氢铵作为氮源,研究在分批培养及流加培养条件下其对螺旋藻生长的影响。结果表明：当培养液中碳酸氢铵浓度小于5mmol/L时,螺旋藻生长正常;碳酸氢铵浓度超过5mmol/L时,螺旋藻生长受到抑制,解体死亡。采用生物量反馈补料的流加策略可以使培养液中螺旋藻生物量达到3.08g/L,产率达到0.26g/(L ·d),藻体中蛋白质及叶绿素含量分别达到65.06%和13.37mg/g,结果证实了碳酸氢铵为氮源高密度培养螺旋藻的可行性。     

无机氮源对红曲霉调控初探

采用以硫酸铵、硝酸铵、硝酸钾、亚硝酸钠4种不同的无机盐作为氮源,探讨其对红曲霉（Monascus）菌株GZUM-25菌落形态、菌丝形态以及色素种类和产量的影响。结果表明,在以硫酸铵为氮源的培养基上,红曲霉菌落的基质菌丝和气生菌丝生长紧密,且气生菌丝在边缘呈放射状,菌丝粗壮、分支多,色素种类多且色价最高为13.5 U/mL;在以硝酸钾为氮源的培养基上,菌落基质菌丝较为疏松,气生菌丝呈毛绒状,菌丝较为粗壮但不具有分支,色素种类单一,色价较高为1.91 U/mL;在以亚硝酸钠为氮源的培养基上,菌落基质菌丝细长且不具有分支,色价最低0.08 U/mL。由此可知,铵盐可以促进红曲霉菌丝的生长发育和次级代谢产物的生成,而亚硝酸盐则有抑制作用。     

Microalgal cultivation in a solution recovered from the low-temperature catalytic gasification of the microalga

Microalgal cultivation in a solution recovered from the low-temperature catalytic gasification of the microalga itself was studied. The growth of Chlorella vulgaris in 75-300-fold diluted recovered solution containing phosphate, magnesium ions and micro-elements was comparable to that in the standard culture medium. It was suggested that C. vulgaris could use ammonium in the recovered solution as its nitrogen source and at the same time could provide a source of biomass which was recycled via gasification.     

氮源对2株海洋微藻生长及脂肪酸合成的影响

以三角褐指藻、扁藻为材料,研究了NaNO3、尿素和NH4Cl等不同氮源对其生长、叶绿素、可溶性糖、可溶性蛋白、干重、总脂及脂肪酸含量的影响。结果表明:以尿素为氮源时,三角褐指藻的生物量、生长速率、叶绿素、可溶性糖、可溶性蛋白的含量最高,NaNO3和NH4Cl次之;以NH4Cl为氮源时,细胞内的总脂含量最高,占干重的47.5℅。与三角褐指藻不同,以NH4Cl为氮源时,扁藻的生物量、生长速率、叶绿素、可溶性糖、可溶性蛋白的含量最高,尿素和NaNO3次之;当以NaNO3为氮源时,细胞内的总脂含量最高,占干重的38.5℅。     

培养条件对蛋白核小球藻生长及油脂合成的影响

以富油蛋白核小球藻为出发藻株,研究自养、异养和混养培养模式对小球藻生物量和油脂含量的影响,以及异养发酵培养基葡萄糖质量浓度、氮源种类及质量浓度对小球藻生长的影响。结果表明,与自养和混养培养模式相比,采用异养发酵方式培养蛋白核小球藻可获得最大的生物量和油脂含量。通过气相色谱法测得异养蛋白核小球藻油主要脂肪酸为棕榈酸(36.07%)、油酸(34.26%)、亚油酸(20.17%)和亚麻酸(6.12%)。经单因素试验优化得到最适蛋白核小球藻生长异养发酵培养基的葡萄糖质量浓度为60 g/L,最适的氮源为酵母粉,质量浓度为4 g/L,在此条件下经192 h发酵,蛋白核小球藻生物量可达12.43 g/L,油脂产量为5.45 g/L。研究结果表明,异养发酵培养获得的蛋白核小球藻油是一种潜在且可再生的新油源。     

C- and N-catabolic utilization of tricarboxylic acid cycle-related amino acids by Scheffersomyces stipitis and other yeasts

Scheffersomyces stipitis and the closely related yeast Candida shehatae assimilated the L-amino acids glutamate, aspartate and proline as both carbon and nitrogen sole sources. We also found this rarely investigated ability in ascomycetous species such as Candida glabrata, C. reukaufii, C. utilis, Debaryomyces hansenii, Kluyveromyces lactis, K. marxianus, Candida albicans, L. elongisporus, Meyerozyma guilliermondii, C. maltosa, Pichia capsulata and Yarrowia lipolytica and in basidiomycetous species such as Rhodotorula rubra and Trichosporon beigelii. Glutamate was a very efficient carbon source for Sc. stipitis, which enabled a high biomass yield/mole, although the growth rate was lower when compared to growth on glucose medium. The cells secreted waste ammonium during growth on glutamate alone. In Sc. stipitis cultures grown in glucose medium containing glutamate as the nitrogen source the biomass yield was maximal, and ethanol concentration and specific ethanol formation rate were significantly higher than in glucose medium containing ammonium as the nitrogen source. Mainly C-assimilation of glutamate but also N-assimilation in glucose-containing medium correlated with enhanced activity of the NAD-dependent glutamate dehydrogenase 2 (GDH2). A Δgdh2 disruptant was unable to utilize glutamate as either a carbon or a nitrogen source; moreover, this disruptant was also unable to utilize aspartate as a carbon source. The mutation was complemented by retransformation of the GDH2 ORF into the Δgdh2 strain. The results show that Gdh2p plays a dual role in Sc. stipitis as both C- and N-catabolic enzyme, which indicates its role as an interface between the carbon and nitrogen metabolism of this yeast.     

三角褐指藻跑道池规模化培养及岩藻黄素积累条件的优化

岩藻黄素是一种广泛存在于海洋藻类的类胡萝卜素,具有清除自由基、抗癌和抗肥胖等生理活性,尤其在抗肿瘤和减肥方面功效突出。三角褐指藻中的岩藻黄素含量丰富,而环境因子及营养因素对微藻生长及岩藻黄素含量有着显著影响。本文以OD值、细胞密度、干重和岩藻黄素含量等为评价指标,研究了不同盐度(20、25、30)、pH值(7.5、8.0、8.5)、氮源种类(尿素、碳酸氢铵)等因素对三角褐指藻户外生长、岩藻黄素积累的影响。研究表明:盐度为20条件下,三角褐指藻获得最大细胞密度6.01×10~6/m L,不同盐度对岩藻黄素含量的增长无明显效果;pH值8.0条件下,三角褐指藻细胞密度及岩藻黄素含量均达到最大值(5.8×10~6/m L和17.79 mg/g);与碳铵相比尿素作为氮源更有利于三角褐指藻的生长,最终细胞密度为4.1×10~6/m L,但以碳铵为氮源条件下,更利于岩藻黄素的积累,岩藻黄素含量达到9.96 mg/g。因此,户外跑道池中的最适培养条件为盐度20、pH值8.0、碳铵为氮源。     

钝顶螺旋藻的硒化培养研究

在培养基中加入亚硒酸钠,对钝顶螺旋藻进行瓶式培养,研究不同加硒浓度、加硒方式对螺旋藻的生长状况及硒积累的影响。结果表明：钝顶螺旋藻中累积的硒随外加硒浓度的增加而增大,但累积系数与浓度的增加成负相关。分次加硒有利于螺旋藻的生长且较一次性加硒获得藻粉的含硒量更高。     

Effect of Sugar and Nitrogen on the Production of Anthocyanin in Cultured Carrot (Daucus carota) cells

ABSTRACT: Callus cultures of carrot, Nentes scarlet - 104 variety, were initiated on MS medium for anthocyanin production. Two anthocyanins, cayanidin-3-lathyroside and cyanidin-3-glucoside, PRESENT in the ratio of 3:1, were identified in the callus cultures. Eight sugars were tried as carbon source for the production of total anthocyanin. The sugars xylose and lactose, although they initiated growth of the green callus, did not initiate anthocyanin pigmentation. Fructose, galactose, and maltose produced less than 1.75% (dry weight basis) anthocyanin though there was growth of the pigmented callus. Glucose and sucrose produced 3.5%. It was observed that 7.5% sucrose in the medium produced maximum amount of anthocyanin (6.5%). Total nitrogen at 70 mM concentration and a 1:4 ratio of ammonium to nitrate yielded maximum cell growth and best anthocyanin production. Modifying the medium it was possible to enhance the production to 6-8%.     

氮源对低温发酵脐橙果酒挥发性成分的影响

为解决低温发酵周期长的问题,并进一步分析其挥发性成分。通过考察不同氮源质量浓度对低温发酵脐橙果酒酒精度和发酵时间的影响,确定氮源添加量,并采用顶空-固相微萃取-气质联用法(HS-SPME-GC-MS)分析其挥发性成分。结果表明,与未添加氮源的空白组相比,添加DAP 0.4 g/L、硫酸铵1.0 g/L、碳酸氢铵1.5 g/L的脐橙果酒酒精度可提高4.6%~5.3%,缩短发酵时间22%~56%。四种低温发酵脐橙果酒共检测出挥发性成分32种,其中高级醇5种、萜烯类化合物10种、中链脂肪酸2种、高级醇乙酸酯3种、中链脂肪酸乙酯7种、醛酮类5种。添加三种氮源的果酒中,萜烯类和乙酸酯类挥发性成分总含量均大于空白组,中链脂肪酸和乙基酯类挥发性成分总含量均小于空白组。其中,添加磷酸氢二铵的果酒中乙酸酯类挥发性成分总含量最高(1.170 mg/L),添加硫酸铵的果酒中萜烯类挥发性成分总含量最高(11.378 mg/L);与另外两种氮源相比,添加磷酸氢二铵的果酒中乙酸-2-苯乙酯、庚酸乙酯等挥发性成分含量相对更高,添加硫酸铵的果酒中D-柠檬烯、乙酸乙酯、癸酸乙酯等挥发性成分含量相对更高,添加碳酸氢铵的果酒中β-水芹烯、4-萜烯醇、辛酸乙酯等挥发性成分含量相对更高。实验证明了氮源对促进脐橙果酒低温发酵具有明显效果,并对挥发性香气成分含量具有一定的促进作用,为脐橙果酒低温发酵工艺的应用提供了理论依据。     

不同基因型烟草对氮素营养响应的差异研究

采用水培盆栽试验,研究了氮素营养对不同基因型烟草幼苗生长的影响.结果表明,不同基因型烟草对氮素营养的响应有明显差异,其硝酸盐还原酶活性、叶绿素含量、根系总吸收面积、氮含量均与氮浓度呈显著或极显著相关.不同基因型间硝酸盐还原酶活性差异显著,其中不耐氮肥基因型8136、红花大金元的硝酸盐还原酶活性明显高于其它品种,其叶绿素含量、根系活力及氮含量也高于其它品种,但其生物量随氮浓度的升高而降低;而较耐氮肥的基因型中烟90、KY17则与之相反.     

液态发酵生产水溶性红曲黄色素的条件优化

以红色红曲菌（Monascus ruber）M-7为实验菌株,以色价和色调为评价指标,对液态发酵生产水溶性红色红曲黄色素的条件进行了研究。结果表明,培养基pH值为3～4时,红色红曲菌M-7能合成大量的水溶性红曲黄色素,发酵液呈鲜艳的黄色色调;培养基最优碳源为2%蔗糖,最优氮源为0.2%硫酸铵;水溶性红色红曲黄色素的色价和色调值随着培养时间的延长先升后降,在第8天时达到最高,色价为5.56 U/mL,OD400 nm/OD510 nm为10.29,OD400 nm/OD470 nm为3.93。     

产油脂小球藻的筛选及其培养基的研究

从7株不同的小球藻中筛选到1株油脂产率较高的普通小球藻C.vulgaris No.1,研究了含氮量不同的培养基对其生长和油脂积累的影响,并用气相色谱法分析其脂肪酸组成.结果表明,高氮培养基有利于C vulgaris No.1的生长,生物量达到1.231 g/L.高氮培养基中的油脂产率最高,达到0.255 g/L,为低氮培养基中的油脂产率的2倍;SE培养基中C.vulgaris No.1的油脂积累也高于低氮培养基.气相色谱分析小球藻主要含十六碳脂肪酸和十八碳脂肪酸,总不饱和脂肪酸含量达80%左右,脂肪酸组成与植物油相似,可作为生产生物柴油的原料.     

不同碳氮组合对三角褐指藻兼养生长及脂质积累的影响

为了探讨兼养培养中不同碳氮组合对三角褐指藻(P.tricornutum)生长及脂质积累的影响,以葡萄糖为碳源,氯化铵为氮源,以不同的碳氮组合兼养培养P.tricornutum,用干重法测定P.tricornutum的生物量,三氯甲烷-甲醇法提取脂质,并采用GC分析脂肪酸组成。结果表明,兼养生长中P.tricornutum基于氯化铵消耗的细胞得率系数随着葡萄糖质量浓度的增加而降低,降低了对氮源的需求;在碳氮组合为葡萄糖质量浓度0.5 g/L,氯化铵浓度1.0 mmol/L的条件下,P.tricornutum生物量达最高,为1.25 g/L,脂质含量可达44.6%,不同碳氮组合对P.tricornutum脂质多不饱和脂肪酸(PUFA)和二十碳五烯酸(EPA)含量的影响不显著。     

Selection of microalgal growth model for describing specific growth rate-light response using extended information criterion

The effects of light intensity and carbon dioxide (CO2) concentration on the growth of the highly CO2-tolerant green alga Chlorococcum littorale were studied in batch cultures. Four mathematical representations were compared for the specific growth rate-light response curve: a rectangular hyperbolic function, Steele's exponential function, a Poisson function and a hyperbolic tangent function. The hyperbolic tangent function, which is commonly used for representing the photosynthesis-light relationship, gave the best fit as evaluated by the extended information criterion (EIC). EIC proved to be applicable as a criterion to this kind of nonlinear model selection problem. Carbon dioxide, the sole carbon source for photoautotrophic growth of this alga, inhibited the growth rate at concentrations higher than pCO2 of 0.02. A substrate inhibition model was successfully used to simulate the relationship between the specific growth rate and CO2 response.     

CONTROL OF AMMONIUM UPTAKE FROM MALT EXTRACT MEDIUM BY SACCHAROMYCES CEREVISIAE

When glucose was used as the energy source in the assay, the perceived ammonium permease activity of Saccharomyces cerevisiae NCYC 1108 remained relatively unchanged in yeast samples taken throughout the main period of fermentation of a malt extract medium. This was in contrast to the earlier work on the fermentation of a defined glucose/salts medium where the specific ammonium permease activity rose rapidly as fermentation proceeded. The present data suggest that the level of ammonium permease activity of cells fermenting a malt extract medium was low due to nitrogen catabolite repression and that inhibition of the carrier system was important in determining the actual rate of uptake from the medium. When ammonium permease in cells from a malt extract medium was assayed using maltose, essentially similar results were obtained. However, the actual values were lower and the results suggested that at some points during the fermentation the ability of the cells to take up maltose during the assay was an important factor in determination of the perceived ammonium permease activity.     

Biological treatment of wastewater discharged from biodiesel fuel production plant with alkali-catalyzed transesterification

The biological treatment of wastewater discharged from a biodiesel fuel (BDF) production plant conducting alkali catalysis transesterification was investigated. BDF wastewater has a high pH and high hexane-extracted oil and low nitrogen concentrations, and inhibits the growth of microorganisms. The biological treatment of BDF wastewater is difficult because the composition of such wastewater is not suitable for microbial growth. To apply the microbiological treatment of BDF wastewater using an oil degradable yeast, Rhodotorula mucilaginosa, the pH was adjusted to 6.8 and several nutrients such as a nitrogen source (ammonium sulfate, ammonium chloride or urea), yeast extract, KH2PO4 and MgSO4.7H2O were added to the wastewater. The optimal initial concentration of yeast extract was 1 g/l and the optimal C/N ratio was between 17 and 68 when using urea as a nitrogen source. A growth inhibitor was also present in the BDF wastewater, and this growth inhibitor could be detected by measuring the solid content in an aqueous phase after the hexane extraction of the wastewater. Microorganisms could not grow at solid contents higher than 2.14 g/l in the wastewater. To avoid the growth inhibition, the BDF wastewater was diluted with the same volume of water. Oil degradation in the diluted BDF wastewater was observed and the best result was obtained under the determined optimal conditions. This treatment system is simple because no controllers, except for a temperature, are necessary. These results suggest that the biological treatment system developed for BDF wastewater is useful for small-scale BDF production plants.     

Altered Patterns of Maltose and Glucose Fermentation by Brewing and Wine Yeasts Influenced by the Complexity of Nitrogen Source

Maltose and glucose fermentations by industrial brewing and wine yeasts strains were strongly affected by the structural complexity of the nitrogen source. In this study, four Saccharomyces cerevisiae strains, two brewing and two wine yeasts, were grown in a medium containing maltose or glucose supplemented with a nitrogen source varying from a single ammonium salt (ammonium sulfate) to free amino acids (casamino acids) and peptides (peptone). Diauxie was observed at low sugar concentration for brewing and wine strains, independent of nitrogen supplementation, and the type of sugar. At high sugar concentrations altered patterns of sugar fermentation were observed, and biomass accumulation and ethanol production depended on the nature of the nitrogen source and were different for brewing and wine strains. In maltose, high biomass production was observed under peptone and casamino acids for the brewing and wine strains, however efficient maltose utilization and high ethanol production was only observed in the presence of casamino acids for one brewing and one wine strain studied. Conversely, peptone and casamino acids induced higher biomass and ethanol production for the two other brewing and wine strains studied. With glucose, in general, peptone induced higher fermentation performance for all strains, and one brewing and wine strain produced the same amount of ethanol with peptone and casamino acids supplementation. Ammonium salts always induced poor yeast performance. The results described in this paper suggest that the complex nitrogen composition of the cultivation medium may create conditions resembling those responsible for inducing sluggish/stuck fermentation, and indicate that the kind and concentration of sugar, the complexity of nitrogen source and the yeast genetic background influence optimal industrial yeast fermentation performance.     

Effect of ammonium concentration on alcoholic fermentation kinetics by wine yeasts for high sugar content

Kinetics of alcoholic fermentation by Saccharomyces cerevisiae wine strains in a synthetic medium with high sugar content were established for different nitrogen initial content and are presented for four strains. The composition of the medium was close to grape must except that the nitrogen source consisted mainly in ammonium and was varied from 120 to 290 mg N/l assimilable nitrogen. The overall nitrogen consumed was also estimated in order to determine nitrogen requirement variability. The effect of assimilable nitrogen was in general greater on sugar consumption rates than on growth and three kinds of effect on sugar consumption rates were observed: (i) existence of an optimal initial nitrogen level for a maximal sugar consumption rate (inhibition if excess), (ii) no effect of nitrogen beyond the intermediary level (saturation), (iii) sugar consumption rate proportional to the initial nitrogen level (activation). In all cases, the amount of consumed nitrogen increased with its initial concentration and so did the fructophilic capacity of the strains. The optimal requirement varied from 0.62 to 0.91 mg N/g of sugars according to different strains. There was no general correlation between the sugar assimilation rates and the nitrogen requirement.