镧对镉离子胁迫下黑曲霉抗氧化酶活性变化的影响

分别应用含有120μmol·L-1 La3+,50μmol·L-1 Cd2+,100μmol·L-1 Cd2+,50μmol·L-1 Cd2++120μmol·L-1 La3+,100μmol·L-1Cd2++120μmol·L-1 La3+的培养基培养黑曲霉,3 d后测定黑曲霉菌球的生物量及其胞内超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、抗坏血酸过氧化物酶(APX)、过氧化物酶(POD)和谷胱甘肽-S-转移酶(GST)活性及脂质过氧化产物(MDA)的变化。结果表明,50和100μmol.L-1Cd2+降低了黑曲霉生物量和五种抗氧化酶活性,并诱导了MDA产物的升高。与Cd2+单一处理比较,50μmol·L-1 Cd2++120μmol·L-1 La3+和100μmol·L-1 Cd2++120μmol·L-1 La3+处理组均不同程度上诱导了五种抗氧化酶活性的升高,并降低了MDA的积累水平。其中,120μmol·L-1 La3+对50μmol·L-1 Cd2+氧化胁迫的缓解效应明显高于100μmol·L-1 Cd2+。因此,La对黑曲霉中低剂量Cd胁迫的缓解效应比较显著,而对高剂量Cd的拮抗效应不明显。     

Studies on Immobilization of Aminoacylase From Aspergillus Oryzae on Macroporous Carriers

Synthesized macroporous cross-linked copolymers of methyl acrylate-divinyl benzene (MA-DVB), acrylamide-N,N'-methylenebisacrylamide (AAM-BIS) and their functionalized products were used for immobilization of aminoacylase from Aspergillus oryzae. Effects of the carrier properties on the activity of immobilized aminoacylase were investigated and effects of substrate concentration, pH, phosphate buffer concentration and temperature on the immobilized aminoacylase were compared with those of the soluble aminoacylase. A column of immobilized aminoaoylase was prepared and used for continuous resolution of N-acyl-DL-methionine; the operational stability of immobilized enzyme was also investigated.     

Resolution of Δ2-isoxazoline-5-carboxylates by a protease fromAspergillus Oryzae providing masked synthons for enantiopure β-aminoalcohols and related structures

Summary A series of racemic ²-isoxazolinecarboxylates have been synthesized and subjected to enzymatic hydrolysis by a protease fromAspergillus oryzae in a two-phase system. Out of these compounds only isoxazoline-5-carboxylates unsubstituted at C-4 were hydrolyzed. Thus, from 3-ethoxycarbonyl-, 3-methyl-, and 3-phenyl-²-isoxazoline-5-carboxylates the corresponding (R)-configurated carboxylic acids are obtained. In contrast, an additional methyl group at C-5 changes the steric course of the hydrolysis to give predominantly the (S)-acid. The enantioselectivities obtained are in the range ofE=5–35.

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Racematspaltung von ²-Isoxazolin-5-carboxylaten mittels einer Protease vonAspergillus oryzae zur Darstellung maskierter Synthons für enantiomerenreine -Aminoalkohole und verwandte Strukturen

Zusammenfassung Zahlreiche ²-Isoxazolincarbonsäureester wurden in racemischer Form dargestellt und einer enzymatischen Hydrolyse mittels einer Protease ausAspergillus oryzae in einem Zwei-Phasen-System unterworfen. Lediglich an C-4 unsubstituierte Isoxazolin-5-carbonsäureester wurden hydrolysiert. Aus 3-Ethoxycarbonyl-, 3-Methyl- und 3-Phenyl-²-isoxazolin-5-carbonsäureestern wurden die entsprechenden (R)-konfigurierten Carbonsäuren erhalten, während eine zusätzliche Methylgruppe an C-5 eine bevorzugte Bildung der (S)-Säure bewirkte. Die erzielten Enantiomerenüberschüsse liegen im Bereich vonE=5–35.

     

Extracellular proteolytic processing ofAspergillus awamori GAI into GAII is supported by physico-chemical evidence

The proportion of glucoamylases, GAI and GAII, in the culture supernatant ofAspergillus awamori fermentations depends on the medium C/N ratio in such a way that the transformation of GAI into GAII is favored by the existence of a surplus of the carbon source in the growth medium. This condition also favors the appearance of the proteolytic activity. The authors report the observation that the shift in the isoenzyme proportion was concomitant to the peak of proteolytic activity. A peptide that may have resulted from the continuous degradation of the GAI C-terminal peptide, Gp-1, was isolated by gel filtration and purified by reversephase chromatography. This peptide matched with the region G¹⁴-A³⁴ of the substrate-binding domain of GAI, thus reinforcing the hypothesis of the extracellular proteolytic processing of GAI.     

Production of a bioflocculant by Aspergillus parasiticus and its application in dye removal

Aspergillus parasiticus was found to produce a bioflocculant with high flocculating activity for Kaolin suspension and water-soluble dyes. Results showed that the carbon and nitrogen sources favorable for the production of the bioflocculant were corn starch and peptone, and an optimal condition of 28 °C, initial pH 5–6 and shaking speed of 150 rpm. The highest flocculating efficiency achieved for Kaolin suspension was 98.1%, after 72 h cultivation. The bioflocculant was mainly composed of sugar (76.3%) and protein (21.6%), and an average molecular weight of 3.2 × 10⁵ Da. X-ray photoelectron spectroscopy (XPS) and Fourier transform infrared (FTIR) spectra showed that amino, amide and hydroxyl groups were present in the bioflocculant molecules. The bioflocculant was effective in flocculating some soluble anionic dyes in aqueous solution, in particular Reactive Blue 4 and Acid Yellow 25 with a decolorization efficiency of 92.4 and 92.9%, respectively. The decolorization efficiency was dependent on the flocculant dosage and solution pH. XPS result shows that the amine groups in the bioflocculant were protonated at pH 5, and thus the positive bioflocculant was attracted to the negatively charged dye molecules. The amino and amide groups in the bioflocculant molecule are believed to play an important role in flocculation from the viewpoint of electrostatic interaction.     

Speradine A, a new pentacyclic oxindole alkaloid from a marine-derived fungus Aspergillus tamarii

A new pentacyclic oxindole alkaloid, speradine A (1), was isolated from the cultured broth of a fungus Aspergillus tamarii, which was separated from driftwood at a seashore in Okinawa. The structure and relative stereochemistry were determined by spectroscopic data and a single crystal X-ray diffraction analysis.     

Effect of media composition and growth conditions on production of β-glucosidase by Aspergillus niger C-6

The hydrolytic activity of fungal originated β-glucosidase is exploited in several biotechnological processes to increase the rate and extent of saccharification of several cellulosic materials by hydrolyzing the cellobiose which inhibits cellulases. In a previous presentation, we reported the screening and liquid fermentation with Aspergillus niger, strain C-6 for β-glucosidase production at shake flask cultures in a basal culture medium with mineral salts, corn syrup liquor, and different waste lignocellulosic materials as the sole carbon source obtaining the maximum enzymatic activity after 5–6 d of 8.5 IU/mL using native sugar cane bagasse. In this work we describe the evaluation of fermentation conditions: growth temperature, medium composition, and pH, also the agitation and aeration effects for β-glucosidase production under submerged culture using a culture media with corn syrup liquor (CSL) and native sugar cane bagasse pith as the sole carbon source in a laboratory fermenter. The maximum enzyme titer of 7.2 IU/mL was obtained within 3 d of fermentation. This indicates that β-glucosidase productivity by Aspergillus niger C-6 is function of culture conditions, principally temperature, pH, culture medium conditions, and the oxygen supply given in the bioreactor. Results obtained suggest that this strain is a potential microorganism that can reach a major level of enzyme production and also for enzyme characterization.     

Citric acid production with alginate bead entrappedAspergillus niger ATCC 9142

Aspergillus niger ATCC 9142 mycelium was entrapped in calcium alginate beads and employed in an air-lift completely stirred reactor for continuous production of citric acid. Maximum yield obtained from 10% (w/v) sucrose was 12 g dm^-3 with about 40% fermentation efficiency. Maximum rate of production 70 mg g^-1 h^-1 was about five times that obtained in classical batch fermentation.     

A new naphthoquinoneimine derivative from the marine algal-derived endophytic fungus Aspergillus niger EN-13

Cultivation of an endophytic fungus Aspergillus niger EN-13 that was isolated from the inner tissue of the marine brown alga Colpomenia sinuosa resulted in the characterization of a new naphthoquinoneimine derivative,namely,5,7-dihydroxy-2-[1-(4- methoxy-6-oxo-6H-pyran-2-yl)-2-phenylethylamino]-[1,4]naphthoquinone.The structure of the new compound was established on the basis of various NMR spectroscopic analyses including 2D NMR techniques,EI-MS,and HR-ESI-MS.This compound displayed moderate antifungal activity.