Bioconversion of pyrimidine by resting cells of quinoline-degrading bacteria

Nine quinoline-degrading bacterial strains were tested for their ability to hydroxylate pyrimidine. All strains converted pyrimidine to uracil via pyrimidine-4-one in a cometabolic process. Quinoline 2-oxidoreductases (QuinORs) were the catalysts of fortuitous pyrimidine hydroxylation. Whereas in most strains the activity of the QuinOR towards pyrimidine was very low compared to its activity towards quinoline, QuinOR in crude extracts from Comamonas testosteroni 63 showed a specific activity of 64 (mU mg protein)-1 with pyrimidine as substrate, compared to a specific activity of 237 (mU mg protein)-1 towards the intrinsic substrate quinoline. Resting cells of Comamonas testosteroni 63 rapidly converted pyrimidine almost stoichiometrically to uracil, which accumulated in the cell suspension. Using an adsorbent resin, uracil was prepared from the supernatant of Comamonas testosteroni 63 resting cells with a yield of > 98%.     

Bioconversion of AHX to AOH by resting cells of Burkholderia contaminans CH-1

Fairy rings are zones of stimulated grass growth owing to the interaction between a fungus and a plant. We previously reported the discovery of two novel plant-growth regulating compounds related to forming fairy rings, 2-azahypoxanthine (AHX) and 2-aza-8-oxohypoxanthine (AOH). In this study, a bacterial strain CH-1 was isolated from an airborne-contaminated nutrient medium containing AHX. The strain converted AHX to AOH and identified as Burkholderia contaminans based on the gene sequence of its 16S rDNA. The quantitative production of AOH by resting cells of the strain was achieved. Among seven Burkholderia species, two bacteria and two yeasts tested, B. contaminans CH-1 showed the highest rate of conversion of AHX to AOH. By batch system, up to 10.6 mmol AHX was converted to AOH using the resting cells. The yield of this process reached at 91%.     

Bioconversion of grape must into modulated gluconic acid production by Aspergillus niger ORS-4.410

AIMS: Analysis of regulators for modulated gluconic acid production under surface fermentation (SF) condition using grape must as the cheap carbohydrate source, by mutant Aspergillus niger ORS-4.410. Replacement of conventional fermentation condition by solid-state surface fermentation (SSF) for semi-continuous production of gluconic acid by pseudo-immobilization of A. niger ORS-4.410. METHODS AND RESULTS: Grape must after rectification was utilized for gluconic acid production in batch fermentation in SF and SSF processes using mutant strain of A. niger ORS-4.410. Use of rectified grape must led to the improved levels of gluconic acid production (80-85 g l(-1)) in the fermentation medium containing 0.075% (NH4)2HPO4; 0.1% KH2PO4 and 0.015% MgSO4.7H2O at an initial pH 6.6 (+/-0.1) under surface fermentation. Gluconic acid production was modulated by incorporating the 2% soybean oil, 2% starch and 1% H2O2 in fermentation medium at continuously high aeration rate (2.0 l min(-1)). Interestingly, 95.8% yield of gluconic acid was obtained when A. niger ORS-4.410 was pseudo-immobilized on cellulose fibres (bagasse) under SSF. Four consecutive fermentation cycles were achieved with a conversion rate of 0.752-0.804 g g(-1) of substrate into gluconic acid under SSF. CONCLUSIONS: Use of additives modulated the gluconic acid production under SF condition. Semi-continuous production of gluconic acid was achieved with pseudo-immobilized mycelia of A. niger ORS-4.410 having a promising yield (95.8%) under SSF condition. SIGNIFICANCE AND IMPACT OF THE STUDY: The bioconversion of grape must into modulated gluconic acid production under SSF conditions can further be employed in fermentation industries by replacing the conventional carbohydrate sources and expensive, energy consuming fermentation processes.     

Bioconversion of waste office paper to gluconic acid in a turbine blade reactor by the filamentous fungus Aspergillus niger

Gluconic acid production was investigated using an enzymatic hydrolysate of waste office automation paper in a culture of Aspergillus niger. In repeated batch cultures using flasks, saccharified solution medium (SM) did not show any inhibitory effects on gluconic acid production compared to glucose medium (GM). The average gluconic acid yields were 92% (SM) and 80% (GM). In repeated batch cultures using SM in a turbine blade reactor (TBR), the gluconic acid yields were 60% (SM) and 67% (GM) with 80-100 g/l of gluconic acid. When pure oxygen was supplied the production rate increased to four times higher than when supplying air. Remarkable differences in the morphology of A. niger and dry cell weight between SM and GM were observed. The difference in morphology may have caused a reduction of oxygen transfer, resulting in a decrease in gluconic acid production rate in SM.     

Synthesis of glucose oxidase and catalase by Aspergillus niger in resting cell culture system

The synthesis of glucose oxidase and catalase by Aspergillus niger was investigated using a resting cell culture system without growth being established. Calcium carbonate induced the synthesis of both enzymes and calcium chloride inhibited it. The optimal pH for the biosynthesis of glucose oxidase and catalase was 6.0 and 5.7, respectively. The effects of other bivalent cations, reductive compounds and metabolic products on enzyme synthesis were also tested. The biosynthesis of glucose oxidase and catalase was promoted by MnCO3, thioglycolic acid, pyroracemic acid and gluconic acid.     

Bioconversion of tea polyphenols to bioactive theabrownins by Aspergillus fumigatus

Theabrownins (TB) are water-soluble phenolic compounds associated with the various health benefits of Pu-erh tea, a post-fermented Chinese dark tea. This work reports on the production of theabrownins from infusions of sun-dried green tea leaves using a pure culture of Aspergillus fumigatus isolated from a solid-state Pu-erh tea fermentation. A theabrownins yield of 158 g kg^?1 sun-dried green tea leaves was obtained in 6 days at 45 °C in an aerobic fermentation. In a 2 l fermenter, the yield of theabrownins was 151 g kg^?1 sun-dried green tea leaves in 48 h of aerobic culture (45 °C, 1 vvm aeration rate, 250 rpm agitation speed). Extracellular polyphenol oxidase and peroxidase of A. fumigatus contributed to this bioconversion. Repeated batch fermentation process was used for producing theabrownins but was less productive than the batch process.     

Biotransformation of dopamine to norlaudanosoline by Aspergillus niger

Norlaundanosoline is a key intermediate in the synthesis of the benzylisoquinoline alkaloids providing the upper isoquinoline portion of the morphinan skeleton. This study evaluates the feasibility of using Aspergillus niger as an in situ biotransformation system to produce norlaudanosoline from dopamine. A. niger was chosen because monoamine oxidase can be readily induced in this organism. Monoamine oxidase catalyzes the conversion of dopamine to 3,4-dihydroxyphenylacetaldehyde. In the presence of dopamine, this aldehyde will then undergo a spontaneous Picket-Spengler condensation to form norlaudanosoline. Fermentation condition to form norlaudanosoline. Fermentation conditions were optimized for the production monoamine oxidase by using a two-stage process consisting of a growth stage and an induction stage. pH control was found to be important, and at pH 4.5 dopamine accumulation in the cells was high as was the level of monoamine oxidase. With pH control at 4.5, up to 21% of the cellular dopamine was converted to norlaudanosoline. It is proposed that with further protein engineering improvements, this system may prove suitable for the in situ bio-transformation of dopamine to norlaudanosoline.     

Degradation of orcinol by Aspergillus niger

Aspergillus niger could utilize orcinol (5-methyl-resorcinol or 3,5-dihydroxytoluene) as the sole source of carbon and energy. In the first step of catabolism A. niger hydroxylates orcinol to form 2,3,5-trihydroxytoluene. Its oxidized form, 2-hydroxy-6-methyl-1,4-benzoquinone, was also formed in the culture medium during growth of this organism. Orcinol-grown cells showed a net increase in the intracellular acetate pool, compared with glucose-grown cells. Cell-free extracts of orcinol-grown cells showed higher activity of orcinol hydroxylase, catechol 1,2-oxygenase, and isocitrate lyase than that of glucose-grown cells. Both orcinol-grown and resorcinol-grown cells exhibit similar respiratory activity on all the substrates checked.     

Retting of Flax by Aspergillus niger

In this study, retting was carried out by Aspergillus niger. The pH, galacturonic acid (GA), and total reducing sugar were determined; the end point was identified by the classic empirical processes and by the maximal GA content of the retting water. The process gave clear and resistent fibers, and the retting time was similar to that of current industrial processes with bacterial enzymes. Control of total acidity was not required, since the pH remained close to neutrality throughout the entire process.     

Biotransformation of glycyrrhizin by Aspergillus niger

Biotransformation of glycyrrhizin by Aspergillus niger was investigated and one new compound (1) and one known compound (2) were isolated and identified from the biotransformation products. These were 7β,15α-dihydroxy-3,11-dioxo-oleana-12-en-30-oic acid (1) and 15α-hydroxy-3,11-dione-oleana-12-en-30-oic acid (2). A biotransformation pathway was proposed from HPLC analyses at different reaction times. The biotransformation by A. niger included two stages: first, the two glucuronic acid residues at the C-3 position of glycyrrhizin were hydrolyzed to produce glycyrrhetic acid; and second, glycyrrhetic acid was oxidized and hydroxylated to compounds 1 and 2.     

Oxalic acid production by Aspergillus niger

Oxalic acid is formed by Aspergillus niger at nearly neutral pH values. In this study the applicability of milk whey as a carbon source was investigated, both in shaking flask experiments and in a stirred tank reactor. The influence of pH on oxalic acid formation showed that the maximum production rate and higher concentration of the product are observed at pH 6. At pH 7 the same production rate was obtained although at a lower oxalic acid concentration. The process was shown to be inhibited by product from an oxalic acid concentration of about 10?kg/m³ and its behaviour was fitted by Luong's equation. In a 10-dm³ strirred tank ferment the stirrer speed was varied in a range from 100 to 600 rpm. At values between 200 and 400 rpm, maximum production rates of oxalic acid of 6.8?kg/m³·d and 6.5?kg/m³·d were reached, respec-tively. A final concentration of 41.4?kg oxalic acid/m³ was reached operating at 400 rpm.     

Oxalic acid production by Aspergillus niger

Aspergillus niger is able to produce a quite high concentration of oxalic acid using sucrose as carbon and energy source. Operating at pH higher than 6 and an enriched N and P medium is necessary in order to conduct the fermentation towards oxalic acid production. A pH?shift technique, operating at acid pH?in the first two days and then setting pH?to 6, allowed the productivity to slightly increase in shaking flasks cultures up to 3.0?kg/m³?·?d, with a final oxalic acid concentration of 29?kg/m³. When operating at more controlled conditions, in a stirred tank, both productivity and oxalic acid concentration were improved (4.1?kg/m³?·?d and 33.8?kg/m³, respectively). However the main drawback of this fermentation is the low yield attained (about 0.3?kg oxalic acid/kg sucrose) because most of glucose, resulting from the hydrolysis of sucrose by the extracellular enzymes secreted at the beginning of the fermentation, is very quickly oxidised to gluconic acid, a process which is favoured at a pH?close to 6. Milk whey was proved to be a very good substrate as it allows oxalic acid to be produced with a similar productivity (2.5?kg/m³?·?d in shaking flasks) giving excellent yields of almost 0.6?kg oxalic acid/kg lactose.     

Cyano-metal complexes uptake by Aspergillus niger

Aspergillus niger was grown in an industrial gold mining solution containing cyano-metal complexes. Gold, silver, copper, iron and zinc were accumulated, and the major mechanism of metal uptake involved metal precipitation on the cell surface. This could be achieved by the extensive application of instrumental analyses combined to the study of the chemical composition of the solutions, before and after contact with the fungus.     

Measurement of glucose transfer to glycerol by Aspergillus niger transglucosidase

An investigation of transglucosidase thermal stability led to the observation that glycerol serves as a glucosyl acceptor. The transferase activity was subsequently measured using two independent assays containing -methyl-D-glucoside (-MG) and -MG + glycerol. The difference between values for digests with and without glycerol was then calculated giving the percentage of transfer to glycerol.