Studies on aspergilli XVII. Effect of nitrogen sources on growth and fruiting

Summary The effect of ammonium, nitrate, and organic nitrogen on growth and sporulation of 18 Aspergilli was examined in a chemically defined medium in surface culture under controlled conditions. All three forms of nitrogen were metabolized by all the Aspergilli tested. Ammonium nitrogen was not good both for growth and fruiting. This was due to the sharp fall in the pH level which resulted due to the rapid utilization of anions of the ammonium nitrogen than cations. The effect of adding succinic acid in the medium containing ammonium nitrogen has been discussed.Good growth of Aspergilli in media containing nitrate nitrogen with the accompanying rise in the pH of the medium showed that these species are capable of reducing nitrate nitrogen to the level of ammonia. The role of succinic acid in the utilization of nitrate nitrogen was investigated. All fungi accomplished good growth on a medium containing asparagine.     

Fungal production of citric acid

Citric acid is the principal organic acid found in citrus fruits. To meet increasing demands it is produced from carbohydrate feedstock by fermentation with the fungus Aspergillus niger and the yeasts of Candida spp. Effect of various fermentation conditions and the biochemistry of citric acid formation by A. niger have been discussed. Commercially citric acid is produced by surface, submerged and solid state fermentation techniques. Recovery of pure acid from fermentation broth is done primarily by precipitation with lime and also by solvent extraction.     

碳源和氮源对5-酮基-葡萄糖酸生成的影响

氧化葡萄糖杆菌Gluconobacter oxydans可以将葡萄糖氧化成葡萄糖酸,并进一步氧化成2-酮基-葡萄糖酸(2KGA)和5-酮基-葡萄糖酸(5KGA),其中5KGA在催化剂的作用下能够转化为L(+)-酒石酸。为了提高5-酮基-葡萄糖酸产量,以仅生成5KGA的氧化葡萄糖杆菌Gluconobacter oxydans HGI-1为出发菌株,研究不同碳源(蔗糖、乳糖、麦芽糖、淀粉、葡萄糖)和有机氮源(酵母浸粉、鱼粉、玉米浆、黄豆饼粉、棉籽饼粉)对5KGA产量的影响。500 mL摇瓶试验结果表明,当葡萄糖浓度为100 g/L时,5KGA产量最高为98.20 g/L;当有机氮源为酵母浸粉、鱼粉和玉米浆,其添加量的蛋白含量为1.60%时,5KGA产量分别为100.20 g/L、109.10 g/L和99.83 g/L,其中,使用鱼粉的5KGA产量最高,使用玉米浆的5KGA产量比酵母浸粉略低。出于经济考虑,文中选择玉米浆作有机氮源,并在5 L发酵罐中进行分批发酵放大试验,5KGA的产量为93.80 g/L,最大生成速率为3.48 g/(L·h),平均生成速率为1.56 g/(L·h)。结果表明,葡萄糖和玉米浆分别为Gluconobacter oxydans HGI-1规模化生产5KGA的最适碳源和氮源,可利用葡萄糖几乎全部(85.93%)转化为5KGA。     

Application of carbon balance to submerged citric acid production by Aspergillus niger

Summary In an air-lift fermenter, the following production was obtained from 125 g sucrose in mineral medium at pH 2.5 : 15.76 g mycelium dry wt, 107.2 g citric acid anhydrous and 0.594 mol CO₂ within 138 h (run I) and 13.72 g mycelium dry wt, 114.28 g citric acid and 0.516 mol CO₂ within 144 h (run II). Initially, the carbon content of consumed sugar and products did not balance. At the end of fermentation, the carbon content of the products was 0.9%–5.5% higher than that of the consumed sugar. For the purpose of the calculations the carbon content in mycelium was accepted as 0.462.The work was a part of Project No 04.11 CPBP, topic No 2.24     

Studies on citric acid production with immobilized Yarrowia lipolytica in repeated batch and continuous air-lift bioreactors

Citric acid was produced from glucose in repeated-batch shake-flask and continuous air-lift cultivations by calcium-alginate-immobilized Yarrowia lipolytica A-101 yeast. The medium composition was systematically studied in a batch system by using experimental design and empiric modelling. The highest citric acid product concentration of 39 g/l was reached with a medium containing 150 g/l of glucose, 0.105 g/l of potassium dihydrogen phosphate, 0.84 g/l of magnesium sulphate and 21 mg/l of copper sulphate (5.2 mg/l of copper). The results were further improved by hardening the alginate carrier beads with glutaraldehyde, and by activation of the immobilized biocatalyst in a nutrient solution. In continuous air-lift bioreactors with varying height-to-diameter ratio the highest productivity of 350 mg/l per hour with a dilution rate of 0.023 l/h and a citric acid product concentration of 12 g/l was reached with a ratio of 3. Correspondence to: H. Kautola     

The influence of type and concentration of the carbon source on production of citric acid by Aspergillus niger

Summary The influence of various carbon sources and their concentration on the production of citrate by Aspergillus niger has been investigated. The sugars maltose, sucrose, glucose, mannose and fructose (in the given order) were carbon sources giving high yields of citric acid. Optimal yields were observed at sugar concentrations of 10% (w/v), with the exception of glucose (7.5%). No citric acid was produced on media containing less than 2.5% sugar. Precultivation of A. niger on 1% sucrose and transference to a 14% concentration of various other sugars induced citrate accumulation. This could be blocked by the addition of cycloheximide, an inhibitor of de novo protein synthesis. This induction was achieved using maltose, sucrose, glucose, mannose and fructose, and also by some other carbon sources (e.g. glycerol) that gave no citric acid accumulation in direct fermentation. Precultivation of A. niger at high (14%) sucrose concentrations and subsequent transfer to the same concentrations of various other carbohydrates, normally not leading to citric acid production, led to formation of citrate. Endogenous carbon sources were also converted to citrate under these conditions. A 14%-sucrose precultivated mycelium continued producing some citrate upon transfer to 1% sugar. These results indicate that high concentrations of certain carbon sources are required for high citrate yields, because they induce the appropriate metabolic imbalance required for acidogenesis.     

甘蔗糖蜜发酵生产柠檬酸

从8株黑曲霉A.niger菌株中筛选获得1株适应甘蔗糖蜜深层发酵柠檬酸的菌株A.niger-94,研究了巴基斯坦糖蜜的预处理方法,结果表明：巴基斯坦糖蜜不同于国产糖蜜,CaO处理法是最适宜的糖蜜种子培养基预处理方法,CaO法＋草酸复合处理则是最适的糖蜜发酵培养基预处理方法,在上述条件下,摇瓶发酵115h左右,与未经处理的糖蜜相比,产酸由3．7g/100ml提高至9．68g/100mg。     

Studies on Choanephoraceae. VIII. Interaction of carbon and nitrogen sources

Summary The effect of different carbon-nitrogen combinations on the growth and sporulation ofBlakeslea trispora, Choanephora cucurbitarum, Choanephora infundibulifera, Choanephora conjuncta, Choanephora heterospora andChoanephora circinans was studied. Both potassium nitrate and sodium nitrite were found to be useless for the present species, even when they were given in combination with different carbon compounds. In some of the cases only a little improvement in the growth was caused by combining poor carbon sources with good nitrogen sources.     

Effect of acetic and trans-aconitic acids on citric acid production by Aspergillus niger.

The effect of acetic and trans-aconitic acids on citric acid production by A. niger at different pH values was studied. The presence of acetic acid at pH 2 prevented spore germination, while it decreased the fungal growth and citric acid production at other pH values. In the presence of trans-aconitic acid the inhibition was less marked at lower than at higher pH values.     

Metabolism of citric acid production by Aspergillus niger: model definition, steady-state analysis and constrained optimization of citric acid production rate

In an attempt to provide a rational basis for the optimization of citric acid production by A. niger, we developed a mathematical model of the metabolism of this filamentous fungus when in conditions of citric acid accumulation. The present model is based in a previous one, but extended with the inclusion of new metabolic processes and updated with currently available kinetic data. Among the different alternatives to represent the system behavior we have chosen the S-system representation within power-law formalism. This type of representation allows us to verify not only the ability of the model to exhibit a stable steady state of the integrated system but also the robustness and quality of the representation. The model analysis is shown to be self-consistent, with a stable steady state, and in good agreement with experimental evidence. Moreover, the model representation is sufficiently robust, as indicated by sensitivity and steady-state and dynamic analyses. From the steady-state results we concluded that the range of accuracy of the S-system representation is wide enough to model realistic deviations from the nominal steady state. The dynamic analysis indicated a reasonable response time, which provided further indication that the model is adequate. The extensive assessment of the reliability and quality of the model put us in a position to address questions of optimization of the system with respect to increased citrate production. We carried out the constrained optimization of A. niger metabolism with the goal of predicting an enzyme activity profile yielding the maximum rate of citrate production, while, at the same time, keeping all enzyme activities within predetermined, physiologically acceptable ranges. The optimization is based on a method described and tested elsewhere that utilizes the fact that the S-system representation of a metabolic system becomes linear at steady state, which allows application of linear programming techniques. Our results show that: (i) while the present profile of enzyme activities in A. niger at idiophase steady state yields high rates of citric acid production, it still leaves room for changes and suggests possible optimization of the activity profile to over five times the basal rate synthesis; (ii) when the total enzyme concentration is allowed to double its basal value, the citric acid production rate can be increased by more than 12-fold, and even larger values can be attained if the total enzyme concentration is allowed to increase even more (up to 50-fold when the total enzyme concentration may rise up to 10-fold the basal value); and (iii) the systematic search of the best combination of subsets of enzymes shows that, under all conditions assayed, a minimum of 13 enzymes need be modified if significant increases in citric acid are to be obtained. This implies that improvements by single enzyme modulation are unlikely, which is in agreement with the findings of some investigators in this and other fields.     

Joint effect of nitrogen and phosphorus concentrations on citric acid production by different strains of Aspergillus niger grown on an effluent

Summary The joint effects of the initial concentration of N and P on the production of citric acid by 11 strains of Aspergillus niger on an effluent are studied by means of orthogonal full-factorial 2² designs. The results allow the definition of a series of empirical equations describing well differentiated types of behaviour. This demonstrates the usefulness of the procedure followed in order to establish selection criteria for the more suitable strains, to determine the tendencies that permit approaching optimum production and evaluate the viability of different culture options characterized by discontinuous feeding of one or another nutrient.     

Continuous production of citric acid in the reciprocating-jet-bioreactor

In a comprehensive study batch and continuous production of citric acid has been investigated. Fermentations in the reciprocation-jet-bioreactor (RJBR) have been carried out with the fungi Aspergillus niger.In the present paper only the results of continuous fermentations are presented. The paper discusses the influence of medium composition in the influent, input of biomass and frequency of reciprocating motion on citric acid production.