A Maltooligosaccharides Producing α-Amylase from Bacillus subtilis SDP1 Isolated from Rhizosphere of Acacia cyanophylla Lindley

Maltooligosaccharides producing amylases are required in the food industry, especially in breadmaking. The Bacillus subtilis strain SDP1 amylase hydrolyses starch to produce maltotriose and maltotetraose along with maltose after prolonged reactions of 5 h. Bacillus subtilis strain SDP1 was isolated from the rhizosphere of Acacia cyanophylla Lindley from the Çukurova region of Turkey. The highest enzyme production was achieved with soluble starch as the carbon and yeast extract as the nitrogen source and at pH 7.0 and 37°C. Under optimized culture conditions, 68.49 U/mL activity was obtained. SDP1 α-amylase had molecular weight of 61 kD. The optimum pH of the enzyme was 7.0 and was highly active at pH ranging from 5.0 to 9.0. The optimum temperature of the crude enzyme was 60°C, and it retained 83% and 74% of its initial activity after 1 h and 2 h incubation periods, respectively, at 50°C. While, Mn⁺² has a stimulatory effect on the activity, Ca⁺², Mg⁺², Na⁺ did not effect the enzyme activity. Fe⁺³, Ni⁺², Cu⁺² and Co⁺² had an inhibitory effect on SDP1 amylase activity.     

Statistical optimization,partial purification,and characterization of coffee pulp β-glucosidase and its application in ethanol production

Extracellular β-glucosidase was produced using coffee pulp as a sole carbon source by Penicillium verrucosum by solid state fermentation and 897.36±59 U/g enzyme activity was obtained. Increase in 2.21-fold of enzyme activity on optimizing the bioprocess parameters by response surface methodology based on central composite rotatable design is illustrated. Maximum production level of 1,991.17 U/g was obtained with optimum values of pH 4.2, moisture 66.8%, and fermentation duration of 56 h. The enzyme was partially purified and the enzyme activity was optimum at 50°C temperature and at pH 6. The metal ions such as Mg²⁺, Zn²⁺, Ca²⁺, K⁺, detergents, and chelator such as EDTA were effective and further increased the β-glucosidase activity. On application of β-glucosidase for simultaneous saccharifiation and fermentation, 3.3% ethanol was obtained. Thus, this study provides insight on exploitation of P. verrucosum for synthesis of of β-glucosidase using coffee pulp which is available abundantly in coffee processing industries.

     

Characterization of an acidophilic and thermostable α‐amylase from Alicyclobacillus sendaiensis NUST

This paper describes the characterization of an acidophilic and thermostable α‐amylase from Alicyclobacillus sendaiensis NUST. The MW of this enzyme was estimated to be 56 kDa by SDS–PAGE. The enzyme was stable over a range of pH from 2.5 to 5.5 with an optimum around 3.5. Maximum activity of the α‐amylase was observed at pH 3.5 and 85°C in the presence of soluble starch as substrate. The enzyme activity was decreased by Mg²⁺, Cu²⁺, Zn²⁺, Al³⁺, K⁺, Li⁺, Ag⁺, urea, EDTA, trichloroacetic acid and Tween 60 and inhibited by Hg²⁺, Ce²⁺ and SDS, whereas the activity was increased by Mn²⁺, DTT, and β‐mercaptoethanol. Ca²⁺and Fe²⁺ did not affect the enzyme activity.     

Gastric H+, K+-ATPase Inhibition,and Antioxidant Properties of Selected Commonly Consumed Vegetable Sources

Oxidative stress and upregulation of gastric H⁺, K⁺-ATPase enzyme activity have been known to cause ulcer pathogenicity for which safer drugs are yet to be identified. Aqueous extracts of seven commonly consumed vegetable sources were screened for inhibition of H⁺, K⁺-ATPase and antioxidant activities. Results indicated that Z. officinale (Ginger) followed by M. arvensis (Pudina) are potent gastroprotective sources with inhibition of H⁺, K⁺-ATPase of IC₅₀ of 18.3 ± 0.7 and 25.2 ± 0.9 μg gallic acid equivalents/ml respectively, which is almost equivalent or better than the known inhibition of H⁺, K⁺-ATPase—Omeprazole (IC₅₀ ?27 μg/ml). Further, all these vegetable extracts showed multi-potent antioxidant activity, such as free radical scavenging, reducing power ability, and inhibition of lipid peroxidation, which are required to inhibit complex steps of ulcerations. On the basis of the absolute amounts and potency of inhibition of H⁺, K⁺-ATPase as well as antioxidant activity of individual phenolic acids, the relative percentage contribution of phenolic acids from different vegetable extracts to both inhibition of H⁺, K⁺-ATPase and antioxidant activity was calculated and data revealed that gentisic and protocatechuic acid contributes significantly to both inhibition of H⁺, K⁺-ATPase and antioxidant activity.     

Effect of gamma irradiation and sodium hydroxide treatment on the debittering of olive (Olea europaea) fruits

The effect of gamma irradiation and sodium hydroxide (NaOH) treatment on the debittering of olive (Olea europaea, var Surrany) fruits was investigated. Fruits were treated with 1, 2 or 3 kGy of gamma irradiation at a dose rate of 669 Gy h^?1. Irradiated and unirradiated fruits were processed with NaOH solution (11 g l^?1) for 3 or 6 h and washed once per day for 3 days. The fruits were then immersed in brine (56 g l^?1 sodium chloride) and stored for 12 months at room temperature. Dissolved organic and inorganic solids, Na⁺, K⁺ and Ca²⁺ contents, pH and electrical conductivity values were determined in the debittering solutions (lye, rinse and washing waters) and brines. Gamma irradiation significantly (p < 0.05) increased the dissolved organic and inorganic solids and the Na⁺ and K⁺ concentrations in the debittering solutions when the fruits were treated with NaOH for 3 h. On the other hand, gamma irradiation had no significant effect on these parameters, except for an increase in K⁺ concentration, when using NaOH solution for 6 h. Gamma irradiation with NaOH treatment for 3 h decreased the concentration of Ca²⁺ in the debittering solutions, whereas irradiation and treatment with NaOH solution for 6 h increased its concentration. © 2003 Society of Chemical Industry     

Purification of New β-Galactosidase from Enterococcus faecium MTCC 5153 with Transgalactosylation Activity

A new β-galactosidase (β-gal) was purified from a lactic acid bacterial strain of Enterococcus faecium MTCC5153 by chromatographic techniques. The purified enzyme had a specific activity of 24.06 U/mg of protein with k _m and V_max values of 2 mM and 18.2 mM/min/mg of protein, respectively. The yield of purified β-gal was 10.65% and estimated molecular weight found to be ～90 kDa, consisting of two homodimeric subunits of 43kDa. The enzyme was stable in pH range of 8.0–9.0 with an optimum pH of 8 and the optimum temperature of 40°C. The enzyme was activated in the presence of metal ions such as Mg⁺², Mn⁺², Ca⁺², K⁺ and Na⁺ and was inhibited by Zn⁺², Co⁺² and Cu⁺². Chemical modifiers (N-bromosuccinamide and Diethylpyro carbonate) inactivated the enzyme indicating the role of tryptophan and histidine moieties for activity. The purified β-gal was able to synthesize oligosaccharides from lactose. This study suggests that the β-gal of Enterococcus faecium MTCC5153 could be applied in dairy industry for hydrolysis of lactose and to improve its digestibility. β-gal of probiotic cultures are of particular interest due to their transgalactosylation properties.     

Characterization of a β-Glucosidase from an Edible Mushroom,Lycoperdon Pyriforme

A β-glucosidase from Lycoperdon pyriforme, a wild edible mushroom, was characterized biochemically. The enzyme showed a maximum activity at pH 4.0 and 50°C when p-nitrophenyl-β-D-glucoside was used as a substrate. K_m and V_max values were calculated as 0.81 mM and 1.62 U/mg protein, respectively. The enzyme activity was conserved about 85% over a broad range of pH (3.0–9.0) at 4°C after 24 h incubation. The activity was fully retained after 60 min incubation at 20–40°C. Na⁺, Li⁺, Mg²⁺, Mn²⁺, Zn²⁺, Co²⁺, Ca²⁺, and Cu²⁺ did not affect the enzyme activity and 0.25% sodium dodecylsulfate inhibited the enzyme activity approximately 76%. Ethylenediamine tetra-acetic acid, phenylmethanesulfonylfluoride, and dithiothreitol showed no or a little negative effect on the enzyme activity. The resistance of the enzyme to some metal ions, chemicals, and ethanol along with the pH stability, can make it attractive for future applications in industry.     

Antioxidant activity of lignans from fringe tree (Chionanthus virginicus L.)

Fringe tree (Chionanthus virginicus L.), a shrub of the eastern part of America, used as a raw material by pharmaceutical industries for cholagoque, diuretic, tonic and the preparation of homeopathic tinctures. The identification of lignans as major antioxidant components and determination of their antioxidant activities are of considerable interest, because of the role they play in pharmacological actions. The potential antioxidant activity of the lignans such as phillyrin, pinoresinol-β-d-glucoside (PDG) and pinoresinol di-β-d-glucoside (PDDG) from root bark of fringe tree (C. virginicus L.) were examined by different antioxidant tests including; 2,2-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) radical scavenging, 1,1-diphenyl-2-picryl-hydrazyl free radical (DPPH^•) scavenging, superoxide anion radical (O₂ ^•-) scavenging, total antioxidant activity by ferric thiocyanate method, reducing activity by Fe³⁺–Fe²⁺ transformation, hydrogen peroxide (H₂O₂) scavenging and ferrous metal (Fe⁺²) chelating activities. Phillyrin, PDG and PDDG, as antioxidants, neutralized the activities of radicals and inhibited the peroxidation reactions of linoleic acid emulsion. The total antioxidant activity was determined according to the ferric thiocyanate method. Butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), α-tocopherol and trolox, which is a water-soluble analogue of tocopherol, were used as the reference antioxidant compounds. Phillyrin, PDG and PDDG showed 67.6, 77.3 and 64.2% inhibition on lipid peroxidation of linoleic acid emulsion, respectively, at the concentration of 20 μg/mL. On the other hand, BHA, BHT, α-tocopherol and trolox exhibited 74.4, 71.2, 54.7 and 20.6% inhibition on peroxidation of linoleic acid emulsion, respectively, at the above mentioned concentration. In addition, phillyrin, PDG and PDDG were effective on DPPH^•, ABTS^•+ and O₂ ^•- scavenging, H₂O₂ scavenging, total reducing power and metal chelating effect on ferrous ions activities. Also, BHA, BHT, α-tocopherol and trolox were used as references antioxidants for these various antioxidant activities.     

Polyphenol oxidase properties,anti-urease,and anti-acetylcholinesterase activity of Diospyros lotus L. (Plum Persimmon)

Diospyros lotus fruit polyphenol oxidase was purified using affinity chromatography, resulting in a 15-fold enrichment in specific activity. The purified enzyme, having 16.5 kDa molecular weight on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, exhibited the highest activity toward 4-methylcatechol. Maximum diphenolase activity was reached at pH 7.0 and 60°C in the presence of 4-methylcatechol. K_m and V_max values were calculated as 3.8 mM and 1250 U/mg protein, respectively. Ascorbic acid was a promising inhibitor with an IC₅₀ value of 0.121 µM. The activity of the purified enzyme was stimulated by Fe²⁺, Sr²⁺, Zn²⁺, and K⁺ and deeply inhibited by Hg²⁺, at 1 mM final concentration. Aqueous extract of Diospyros lotus L. fruit showed strong substantial urease and acetylcholinesterase inhibition, with IC₅₀ values of 1.55 ± 0.05 and 16.75 ± 0.11 mg/mL, respectively.     

Synthesis of Poly(Methacrylic Acid-)Starch Graft Copolymers Using Mn-IV-Acid System

When starch was treated with KMnO₄ solution, MnO₂ was deposited overall the starch. The amount of MnO₂ deposited relied on the KMnO₄ concentration. Subjecting the MnO₂-containing starch to a solution consisting of monomer (methacrylic acid, MAA) and acid (citric, tartaric, oxalic or sulphuric acid) resulted in formation of poly(MAA)-starch graft copolymer. The magnitude of grafting, expressed as meq. -COOH/100 g starch, was determined by amount of MnO₂ deposited, MAA concentration, temperature and duration of polymerization as well as kind and concentration of the acid. Incorporation of cations such as Fe⁺³, Cu⁺² and Li⁺¹ had a significant effect on grafting. The highest extent and rate of grafting were obtained with citric acid and the least with sulphuric acid; tartaric acid stood in-mid-way position. The magnitude of grafting increased as the acid concentration increased till a certain concentration beyond which grafting levelled off. Similar trend was observed when the magnitude of grafting was related to the amount of MnO₂ deposited. The extent and rate of grafting increased by raising the polymerization temperature from 30° to 50°C then decreased by raising the temperature further from 60° to 70°C. On the other hand, grafting enhanced significantly by addition of Fe⁺³, Cu⁺² or Li⁺¹ and followed the order Fe⁺³ > Cu⁺² > Li⁺¹. A tentative mechanism for grafting of starch with MAA using MnO₂-acid system was elucidated.     

Purification and Characterization of Polyphenol Oxidase from Hemşin Apple (Malus communis L.)

The polyphenol oxidase (PPO) enzyme was purified and characterized from Hem?in Apple (Malus communis L.), which was organically grown in Hem?in, in the Rize province of Turkey. Enzyme (PPO) activation was determined with catechol substrate. Apples were homogenized with homogenate buffer (pH 8.5). This process was followed by precipitation with (20–80%) saturated solid (NH₄)₂SO₄ and dialysis. Finally, purification with DE52-Cellulose ion-exchange and Sephadex G-25 columns was performed. Experiments were performed at an optimum pH (5.5) and optimum temperature (30–40°C). The kinetic and thermal parameters Km (3.40 mM), Vmax (333.3 EU/mL.min), Ea (3.57 kcal), ?H (2.968 kcal/mol), Q₁₀ (1.33), k_cat (24.57 min^?1) and V₀ (7.2x10³ mM^?1.min^?1) were assessed. Additionally, the effects of Mg²⁺, Pb ²⁺, Fe²⁺, Fe³⁺, Cd²⁺, Cu²⁺, Zn²⁺, Co²⁺, Al³⁺, Mn²⁺ and Na⁺ on enzyme activity was recorded, and the IC₅₀ values, K_? values and inhibition types were determined.     

PURIFICATION AND PROPERTIES OF CYCLODEXTRIN GLUCANOTRANSFERASE FROM AN ALKALOPHILIC BACILLUS sp. 7-12

Cyclodextrin glucanotransferases (EC 2.4.1.19) (CGTase) are industrially important enzymes for production of cyclodextrin (CD) from starch. γ‐CD yield of CGTase from alkalophilic Bacillus species is usually much lower than β‐CD, while from alkalophilic Bacillus sp. 7‐12. γ‐CD yield is close to β‐CD. A CGTase from alkalophilic Bacillus sp. 7‐12 was purified and characterized. When purified by ammonium sulfate fractionation, DEAE‐cellulose column chromatography and Sepharose CL‐6B column chromatography, the enzyme obtained consisted of a single band that did not dissociate into subunits by SDS polyacrylamide gel electrophoresis. Molecular weight of the purified enzyme was determined to be 69,000 Da by SDS‐PAGE. The enzyme showed a K_mof 1.24 mg/mL and V_max0.101 µM/min when potato starch was used as substrate. The enzyme was stable below 70C with an optimum activity at 60C, and stable at pH range 6–10 with an optimum pH at 8.5. The enzyme activity was strongly inhibited by Ag⁺, Cu²⁺, Mg²⁺, Al³⁺, Co²⁺, Zn²⁺, Fe²⁺and slightly inhibited by Sn²⁺, Mn²⁺. The ions Ca²⁺and K⁺, EDTA and DTT had no influence on the enzyme activity.     

Antimicrobial Activity of Malting Barley Grain Thaumatin‐Like Protein Isoforms,S and R

Two basic proteins were isolated to homogeneity from malting barley (Hordeum vulgare L.) grain. Proteins were identified as members of a Thaumatin‐Like Protein (TLP) family, by Western blot. Isoforms, assigned as TLP‐S and TLP‐R, have slightly different mobility at about 22 and 27 kDa in nonreducing and reducing conditions, and pI values of 9.5 and 9.4, respectively. The antifungal potency of malting barley grain TLPs isoforms was examined on Micrococcus lysodeikticus, Saccharomyces cerevisiae, Candida albicans and plant pathogen Fusarium sporotrichioides growth in vitro. It was found that that IC₅₀ value for TLP‐S was two fold higher. Antibacterial and antifungal activities of both isoforms were completely abolished by divalent (Ca²⁺, Mn²⁺, Mg²⁺) and monovalent (K⁺) cations, at concentrations approximating physiological ionic strength and higher. Glucanase activity was not observed; neither TLP‐S nor TLP‐R digested glucan. On the basis of these results, the importance of TLP for barley grain protection against fungal diseases has been discussed together with the mechanism of antimicrobial action.     

Purification and Characterization of β-Glucosidase from Aspergillus niger

Aspergillus niger, an isolate of soil contaminated with effluents from cotton ginning mill was grown in Czapek-Dox medium containing sawdust, Triton-X 100 and urea for production of an extracellular β-glucosidase. β-Glucosidase enzyme was purified (86-fold) from culture filtrate of A. niger by employing ammonium sulphate precipitation and gel filtration on sephadex G-75. The molecular mass of the purified enzyme was estimated to be 95 kDa by sodium dodecyl sulphate polyacrylamide gel electrophoresis. The enzyme had an optimal activity on p-nitrophenyl β-D-glucopyranoside at 50°C and pH 5.0. The K_m and V_max of the enzyme on p-nitrophenyl β-D-glucopyranoside at 50°C and pH 5 were 8.0 mM and 166 µmol/min/mg of protein, respectively. The enzyme could hydrolyze cellobiose and lactose but not sucrose. Heavy metals like Hg²⁺, Al³⁺, and Ag⁺ inhibited the activity, whereas Zn²⁺ and detergents such as Triton-X 100 and Tween-80 increased the activity at 0.01%. The enzyme activity increased in the presence of methanol and ethanol.     

Effect of Ethylenediaminetetraacetic Acid (EDTA) and Metal Ions on Growth of Staphylococcus aureus 196E in Culture Media

The effects of ethylenediaminetetraacetic acid (EDTA) and its salts, alone or in combination with Mg²⁺, Fe³⁺, Zn²⁺ or Ca²⁺, on growth of S. aureus strain 196E, were studied. EDTA or its Na⁺ or K⁺ salts (0.8–1.7 mM), but not its Ca²⁺ or Fe³⁺ salts, inhibited cell growth in Brain Heart Infusion broth. Bacteriostatic effects in Casamino Acids-Yeast Extract (CasY) broth were produced by 0.17 mM EDTA. Addition of Fe³⁺, Zn²⁺, or Ca²⁺ to EDTA-containing broth eliminated inhibition at an EDTA: cation molar ratio of 1:1, while Mg²⁺ required a sevenfold concentration. Data suggest that EDTA exerts its inhibitory effect by chelating calcium and/or other essential cations which form complexes with comparable stability constants with EDTA.     

Purification and some properties of α-amylase from post-harvest Pachyrhizus erosus L. tuber

α-Amylase, a starch splitting enzyme, was purified to homogeneity from post-harvest Pachyrhizus erosus L. tuber by successive chromatography on DEAE- and CM-cellulose columns. Purification achieved was 110 fold from the crude extract with a yield of 22.8%. SDS-PAGE showed a molecular weight of 40 kDa for the enzyme. The enzyme is of α-type as it lost total activity in the presence of EDTA, a chelating agent. It is a glycoprotein that contains 2.6% sugar as estimated by the phenol-sulfuric acid method. The enzyme displayed optimum activity at pH 7.3 and 37 °C with an apparent K_m value of 0.29% for starch as substrate. The enzyme was strongly inhibited by Cu²⁺, Fe²⁺ and Zn²⁺, moderately by Li²⁺, Hg⁺ and Cd²⁺ and slightly by Ag⁺, Mg²⁺ and K⁺. Calcium ion almost doubled the activity whereas Fe³⁺, Mn²⁺ and Na⁺ enhanced it appreciably.     

Kinetic Study of the Quenching Reaction of Singlet Oxygen by Common Synthetic Antioxidants (tert-Butylhydroxyanisol, tert-di-Butylhydroxytoluene, and tert-Butylhydroquinone) as Compared with α-Tocopherol

ABSTRACT: Effects of synthetic phenolic antioxidants (BHA, BHT, and TBHQ) on the methylene blue (MB) sensitized photooxidation of linoleic acid as compared with that of α‐tocopherol have been studied. Their antioxidative mechanism was studied by both ESR spectroscopy in a 2,2,6,6‐tetramethylpiperidone (TMPD)‐methylene blue (MB) system and spectroscopic analysis of rubrene oxidation induced by a chemical source of singlet oxygen. Total singlet oxygen quenching rate constants (k_ox?Q+k_q) were determined using a steady state kinetic equation. TBHQ showed the strongest protective activity against the MB sensitized photooxidation of linoleic acid, followed by BHA and BHT. TBHQ (1 × 10^?3 M) exhibited 86.5% and 71.4% inhibition of peroxide and conjugated diene formations, respectively, in linoleic acid photooxidation after 60‐min light illumination. The protective activity of TBHQ against the photosensitized oxidation of linoleic acid was almost comparable to that of α‐tocopherol. The data obtained from ESR and rubrene oxidation studies clearly showed the strong singlet oxygen quenching ability of TBHQ. The k_ox?Q+k_q of BHA, BHT, and TBHQ were determined to be 3.37 × 10⁷, 4.26 × 10⁶, and 1.67 × 10⁸ M^?1 s^?1, respectively. The k_ox?Q+k_q of TBHQ was within the same order of magnitude of that of α‐tocopherol, a known efficient singlet oxygen quencher. There was a high negative correlation (r² = ?0.991) between log (k_ox?Q+k_q) and reported oxidation potentials for the synthetic antioxidants, indicating their charge‐transfer mechanism for singlet oxygen quenching. This is the 1st report on the kinetic study on k_ox?Q+k_q of TBHQ in methanol as compared with other commonly used commercial synthetic antioxidants and α‐tocopherol.     

Salt-induced modulation in inorganic nutrients, antioxidant enzymes, proline content and seed oil composition in safflower (Carthamus tinctorius L.)

BACKGROUND: Safflower (Carthamus tinctorius L.) has gained considerable ground as a potential oil‐seed crop. However, its yield and oil production are adversely affected under saline conditions. The present study was conducted to appraise the influence of salt (NaCl) stress on yield, accumulation of different inorganic elements, free proline and activities of some key antioxidant enzymes in plant tissues as well as seed oil components in safflower. Two safflower accessions differing in salt tolerance (Safflower‐33 (salt sensitive) and Safflower‐39 (salt tolerant)) were grown under saline (150 mmol L^?1) conditions and salt‐induced changes in the earlier‐mentioned physiological attributes were determined. RESULTS: Salt stress enhanced leaf and root Na⁺, Cl^? and proline accumulation and activities of leaf superoxide dismutase, catalase and peroxidase, while it decreased K⁺, Ca²⁺ and K⁺/Ca²⁺ and Ca²⁺/Na⁺ ratios and seed yield, 100‐seed weight, number of seeds, as well as capitula, seed oil contents and oil palmitic acid. No significant effect of salt stress was observed on seed oil α‐tocopherols, stearic acid, oleic acid or linoleic acid contents. Of the two safflower lines, salt‐sensitive Safflower‐33 was higher in leaf and root Na⁺ and Cl^?, while Safflower‐39 was higher in leaf and root K⁺, K⁺/Ca²⁺ and Ca²⁺/Na⁺ and seed yield, 100‐seed weight, catalase activity, seed oil contents, seed oil α‐tocopherol and palmitic acid. Other attributes remained almost unaffected in both accessions. CONCLUSION: Overall, high salt tolerance of Safflower‐39 could be attributed to Na⁺ and Cl^? exclusion, high accumulation of K⁺ and free proline, enhanced CAT activity, seed oil α‐tocopherols and palmitic acid contents. Copyright © 2011 Society of Chemical Industry     

Characterization of plasma membrane H+-ATPase from salt-tolerant yeast Candida versatilis

Plasma membrane was isolated from the salt-tolerant yeast Candida versatilis and the ATPase in plasma membrane was characterized. The ATPase was a typical H⁺-ATPase with similar properties to the Saccharomyces cerevisiae and Zygosaccharomyces rouxii enzymes. It was reacted with antibody (IgG) raised against S. cerevisiae plasma membrane H⁺-ATPase. The ATPase activity was not changed by adding NaCl and KCl to the assay solutions, but was increased by NH, especially by ammonium sulfate. In vivo stimulation of ATPase activity was observed by the addition of NaCl into the culture medium, as observed in Z. rouxii. No in vivo activation of H⁺-ATPase by glucose metabolism was observed in C. versatilis cells and the activity was independent of the growth phase, like Z. rouxii and unlike S. cerevisiae cells.     

Optimization of Trace Metals Concentration on Citric Acid Production by Aspergillus niger NRRL 2001

Citric acid is nowadays produced by submerged fermentation of Aspergillus niger. The process yield depends on the composition of the medium, as well as on the microorganism strain. In this work, the effect of Fe⁺³, Zn⁺², and Mn⁺² on citric acid production by A. niger NRRL 2001 is presented. The culture medium composition was glucose (120 g/L) KH₂PO₄ (1.0 g/L); K₂HPO₄ (1.0 g/L), MgSO₄.7H₂O (0.5 g/L), (NH₄)₂SO₄ (3.0 g/L). The ions Fe⁺³, Zn⁺², and Mn⁺² had their concentrations changed according to an experimental design. The experiments were carried out in an orbital shaker at 200 rpm and 30°C. The strain produced an extracellular polysaccharide that was also quantified. The optimum experimental condition was found using 7.0 mg/L of Fe⁺³ and 6.5 mg/L of Zn⁺² in absence of Mn⁺². No oxalic acid formation was observed using this experimental condition. Metal contents were not significant for the production of the polysaccharide. The highest production rate (2.95 g L⁻¹ day⁻¹) was reached after 10 days of fermentation. After this period, the productivity decreased slightly. In 20 days, the citric acid production rate (2.44 g L⁻¹ day⁻¹) was 82% of the highest productivity. The conversion into citric acid increased continuously, yielding 45.8% in 20 days of fermentation.