• The Korean Journal of Mycology
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• v.41 no.2
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• pp.132-135
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• 2013

Black Aspergillus is important fungus for oriental fermentation industry. Black Aspergillus was frequently isolated from Korean traditional Meju, a fermented soybean starting material for soy sauce and soybean paste. Thirty three strains were isolated from 98 finished Meju collected in various regions of Korea from 2008 to 2011, and 21 strains were isolated from in-process Meju at various farms from 2010 to 2011. The isolated black Aspergillus were identified using DNA sequences of partial ${\beta}$-tubulin and calmodulin genes. Of 54 black Aspergillus strains, 14 strains were identified as A. luchuensis and the others were composed of A. niger (n = 21), A. tubingensis (n = 10), and A. welwitschiae (n = 9).

• Proceedings of the Korean Society for Applied Microbiology Conference
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• 1975.12a
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• pp.182.2-182
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• 1975

자연계에서 분리한 유기산 생성균 8주 가운데 구연산 생성능력이 강한 1주를 선정하여 이것이 Aspergillus niger임을 확인하였다. 이 균주를 Sakaguchi's medium 으로 14일 동안 진 탕배양함으로써 17g/l의 구연산이 생산되어, 같은 조건에서 12g/l를 생성하는 기존 균주 A. usamii보다 구연산 생성율이 높았으므로 이 A. niger 사용 균주로 결정하였다.(중략)

• Mycobiology
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• v.31 no.4
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• pp.221-225
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• 2003

Effect of ethanolic extracts of Lawsonia inermis, Azadirachta indica, Vinca rosea, Tagetes patula, Ocimum sanctum, Colocasia antiquorum, Adhatoda vasica, Moringa oleifera, Datura metel and Curcuma longa leaf on conidial germination, mycelial growth and sporulation of Aspergillus flavus, A. niger and A. fumigatus were examined. The conidial germination of A. flavus and A. fumigatus were most inhibited by the extract of L. inermis, while that of A. niger was inhibited by A. indica. Other tested plant extracts have a good effect on conidial germination on the selected fungi. The highest mycelial growth of A. flavus(37 mm) was found in V. rosea, but in case of A. niger and A. fumigatus it(38 and 39 mm) was found in D. metel. The lowest(4, 9 and 6 mm) respectively mycelial growth of these fungi found in L. inermis. The highest sporulation($75{\times}10^4/ml$) of A. flavus was counted in V. rosea, but in case of A. niger and A. fumigatus those($45{\times}10^4$ and $55{\times}10^4/ml$) were in D. metel and the lowest($5{\times}10^4,\;12{\times}10^4\;and\;9{\times}10^4/ml$) respectively sporulation of these fungi counted in L. inermis plant extract medium.

• Journal of Applied Biological Chemistry
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• v.57 no.4
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• pp.313-320
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• 2014

Black liquor (BL) is a by-product of rice straw pulping process. It is a low costs raw material for production value-adding proteins and enzymes, which has been paid more and more attention to reduce its environmental pollution. Mixed cultures of micelial fungi, Trichoderma reesei Northern Regional Research Laboratory (NRRL)11236, Trichoderma reesei NRRL 6165 and Aspergillus niger strains NRC 5A, NRC 7A, and NRC 9A were evaluated for their ability to produce xylanase using crude hemicellulose (CHC) prepared from BL and peat moss as an inert support under solid state fermentation (SSF). The most potent strains, A. niger NRC 9A (818.26 U/g CHC) and T. reesei NRRL 6165 ($100.9{\pm}57.14$ U/g CHC), were used in a mixed culture to enhance xylanase production by co-culturing under SSF. In the mixed culture, xylanase production ($1070.52{\pm}12.57$ U/g CHC) was nearly1.3 and 10.6-fold increases over the activities attained in their monocultures, A. niger NRC 9A and T. reesei NRRL 6165, respectively. Optimization of the culture parameters of the mixed culture SSF process, concentration of ammonium sulfate and corn steep liquor, CHC/peat moss ratio, inoculum size and ratios of the two strains, initial pH value, initial moisture content and incubation time, exhibited a significant increase ($2414.98{\pm}84.02$ U/g CHC) in xylanase production than before optimization.

• KSBB Journal
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• v.9 no.1
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• pp.55-62
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• 1994

Glucose oxidase(EC 1.1.3.4) was purified to electrophoretic homogeneity from Aspergillus niger by a combination of ammonium sulfate fractionation, ion exchange chromatography, and ultrafiltration. Two active fractions A and B, of glucose oxidase were obtained from the hydrophobic chromatography on phenyl sepharose CL-4B. The enzyme A and B were glycoproteins with the same denatured molecular weight of 78, 000 and had specific activities of 2, 191 and 1, 273-units/mg proteins, respectively. But the two enzymes showed differences in native molecular weight that was measured by HPLC gel filteration, maximum absorbtion wavelength and isoelectric point. The enzyme A oxidized $\beta$-D-glucose only and was resistant to sodium dodecyl sulfate. Activity optimum was found at $30^{\circ}C$ and pH 3.5. Also the enzyme A was inhibited greatly by $Hg^{2+}$(10mM). The results of chemical modification experiments suggested that cysteine and cystine residues might be involved in the active site of the enzyme A.

• Mycobiology
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• v.30 no.3
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• pp.160-165
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• 2002

Aspergillus niger has been used as a host to express many heterologous proteins. It has been known that the presence of an- abundant protease is a limiting factor to express a heterologous protein. The protease deficient mutant of A. niger was obtained using UV-irradiation. A total of $1{\times}10^5$ spores were irradiated with $10{\sim}20%$ survival dose of UV, 600 $J/m^2$ at 280 nm, and the resulting spores were screened on the casein-gelatin plates. Ten putative protease deficient mutants showing the reduced halo area around colonies were further analyzed to differentiate the protease deficient mutant from other mutant types. Among ten putative mutants, seven mutants showed significant growth defect on nutrient rich medium and two mutants appeared to be the secretory mutants, which resulted in the impaired secretion of extracellular proteins including proteases. A mutant $pro^--20$ showed reduced halo zone without any notable changes in growth rate. In addition, the starchdegrading and glucose oxidase activities in the culture filtrate of $pro^--20$ mutant showed the similar range as that of the parental strain, which suggested that the $pro^--20$ mutant ought to be the protease deficient mutant rather than a secretory mutant. The reduced proteolytic activity of the $pro^--20$ was demonstrated using SDS-fibrin zymography gel. The reduced extracellular proteolysis was quantified by casein degradation assay and, comparing with the parental strain, less than 30% residual extracellular protease activity was detected in the culture filtrate of the $pro^--20$ mutant. The bio-activity of an exogenously supplemented hGM-CSF(human Granulocyte-Macrophage Colony Stimulating Factor) in the culture filtrate of $pro^--20$ mutant was detected until eight times more diluted preparations than that of the parental strain.

• Asian-Australasian Journal of Animal Sciences
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• v.19 no.9
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• pp.1314-1321
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• 2006

The objective of this work was to study the effect of mixed culture solid substrate fermentation of C. tropicalis ZD-3 with A. niger ZD-8 on detoxification of cottonseed meal (CSM), and to investigate the effect of fermentation period, proportion of CSM in substrate, sodium carbonate, minerals and heat treatment on the reduction of free gossypol levels during mixed culture solid substrate fermentation of CSM. Experiment 1: Three groups of disinfected CSM substrate were incubated for 48 h after inoculation with either of the fungi C. tropicalis ZD-3, A. niger ZD-8 or mixed culture (C. tropicalis ZD-3 with A. niger ZD-8). One non-inoculated group was used as the control. Levels of initial and final free gossypol (FG), CP and in vitro CP digestibility were assayed. The results indicated that mixed culture fermentation was far more effective than single strain fermentation, which not only had higher detoxification rate, but also had higher CP content and in vitro digestibility. Experiment 2: CSM substrates were treated according to experimental variables including fermentation period, proportion of CSM in substrate, sodium carbonate, minerals and heat treatment, Then, the treated CSM substrates were inoculated with mixed culture (C. tropicalis ZD-3 with A. niger ZD-8) and incubated at $30^{\circ}C$ for 36 h in a 95% relative humidity chamber. After fermentation ended, FG and CP content of fermented CSM substrate was assayed. The results showed that the appropriate fermentation period was 36 h, and the optimal proportion of CSM in substrate was 70%. Addition of sodium carbonate to CSM substrate was beneficial for fermentative detoxification. Heat treatment could facilitate fermentative detoxification, and supplementation with minerals was instrumental in reducing gossypol levels during mixed culture solid substrate fermentation of CSM.

• Journal of Microbiology and Biotechnology
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• v.34 no.2
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• pp.330-339
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• 2024

Corn cobs were fermented with Aspergillus niger to produce soluble dietary fiber (SDF) of high quality and excellent food safety. In this work, the fermentation process was optimized by single-factor test and response surface methodology (RSM). The optimal fermentation conditions were determined to be a material-liquid ratio of 1:30, an inoculum concentration of 11%, a temperature of 32℃, a time of 6 days, and a shaking speed of 200 r/min. Under these conditions, the SDF yield of corn cob increased from 2.34% to 11.92%, and the ratio of soluble dietary fiber to total dietary fiber (SDF/TDF) reached 19.08%, meeting the requirements for high-quality dietary fiber (SDF/TDF of more than 10%). Scanning electron microscopy (SEM) and Fourier-transformed infrared spectroscopy (FT-IR) analysis revealed that the fermentation effectively degraded part of cellulose and hemicellulose, resulting in the formation of a loose and porous structure. After fermentation the water swelling capacity, water-holding capacity, and oil-holding capacity of the corn cob SDF were significantly improved and the adsorption capacity of glucose, cholesterol, and nitrite ions all increased by more than 20%. Moreover, the total phenolic content increased by 20.96%, which correlated with the higher antioxidant activity of SDF. Overall, the fermentation of corn cobs by A. niger increased the yield and enhanced the functional properties of dietary fiber (DF) as well.

• 한국생물공학회:학술대회논문집
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• 2001.11a
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• pp.89-92
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• 2001

Aspergillus niger has been used as a host system to express many heterologous proteins. It has various advantages over other expression systems in that it is a small eukaryotic GRAS (Generally Recognized aS Safe) organism with a capacity of secreting large amount of foreign proteins. However, it has been known that the presence of an abundant protease is a limiting factor to express a heterologous protein. The proteases deficient mutants of A. niger were obtained using UV -mutagenesis. A total of 1 ${\times}$ $10^5$ spores were irradiated with 10-20% survival dose of UV, 600J/M2 at 280nm, and the resulting spores were screened on the casein -gelatin plates. Ten putative protease deficient mutants were further analyzed on the starch plates to differentiate the pro from the secretory mutant. An endogenous extracellular enzyme, glucose oxidase, was also examined to confirm that the mutant phenotype was due to the proteases deficiency rather than the mutation in the secretory pathway. The reduced proteolytic activity was measured using SDS-fibrin zymography gel, casein degradation assay, and bio-activity of a supplemented hGM -CSF (human Granulocyte-Macrophage Colony Stimulating Factor). Comparing with the wild type strain, less than 30 % of proteolytic activity was observed in the culture filtrate of the protease deficient mutant (pro -20) without any notable changes in cell growth and secretion.

• KSBB Journal
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• v.11 no.3
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• pp.270-275
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• 1996

Heterologous expression of glucose oxidase gene using recombinant yeast has been carried out. Polymerase chain reaction was conducted to obtain the gene encoding glucose oxidase from Aspergillus niger and sequence comparison indicated the cloned 1.9kb DNA fragment appeared to be the glucose oxidise structural gene containing a signal sequence for extracellular location. Transforming shuttle vector was constructed with YEp352 to express the cloned glucose oxidase gene under the control of either GAL1 or GAL10 promoter. Plate assay of recombinant yeasts has shown that GAL1 promoter was more effective in yielding glucose oxidise than GAL10 promoter. Among the five different concentrations of galactose tried, 1% galactose showed the highest induction of glucose oxidase. Cellular localization experiment of recombinant enzyme using spheroplast revealed that most of enzymes (80%) were secreted into culture media in contrast to A. niger. There is no difference in heat-stability of recombinant enzyme up to $50^{\circ}C$ compared to the glucose oxidase from A. niger However, a dramatic reduction of enzyme activity was observed in both enzymes at $60^{\circ}C$.