• Journal of Microbiology and Biotechnology
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• 제11권6호
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• pp.1066-1070
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• 2001

The PhyA gene, encoding myo-inositol hexakisphosphate phosphohydrolase in Aspergillus niger var. awamori (wild-type), was cloned and sequenced. The cDNA was overexpressed by a multicopy gene expression system in Pichia pastoris KM71. Recombinant, wild-type and commercial phytase from Aspergilus ficuum NRRL 3135 (Natuphos) were purified. The PhyA gene of Aspergillus niger var awamori showed perfect homology to the phytase of Aspergillus ficcum and $97\%$ homology to A. niger var awamori (L02421). Wild-type phytase was highly glycosylated and more thermostable than the other two, while deglycosylated farms of three phytases showed identical molecular weight, 507 kDa. After heating at $80^{\circ}C$, wild-type, commercial, and recombinant phytases retained $57\%, 32%,\;and\;8\%$ of their original activities, respectively. In conclusion, glycosylation plays a key role in the thermostability of phytase and its enzymatic characterization.

• 미생물학회지
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• 제14권1호
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• pp.17-24
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• 1976

Celluloytic microorgnasims were isolated form the various sources and four of them were identified as Trichoderma roningi, Aspergillus niger, Penicillium chrysogenum, and Streptomyces sp. The induction of extracellular5 cellulase of these species in the liquid culture media containing carboxymethylcellulose (CMC) or Avicel as inducer showed that CMC was a better effective inducer for the production of CMCase(Cx cellulase component) as well as Avicelase(C$_{1}$ cellulase component) than Avicel. It is believed that certain hydrolysis products of cellulose(CMC) could serve as an inducer for an enzyme synthesis. In T. roningi, Asp. niger, and Strptomyces sp., the optimum temperature of CNCase on CMC-culture medium was 50.deg. but temperature around 40.deg.C was found to be optimum for the activities of CMCase prepared from P.ehrysogenum. The optimum temperature for Avicelase activitiles on Avicel-culture media of T. roningi and P. chrysogenum was $40{\circ}C$ whereas temperature $50{\circ}C$ was found to be optimum for Avicelase from A.niger and Streptomyces sp. The optimal activities of these CNCase and Avicelase prepared from. T. ronigi, Pen.chrysogenum and Streptomyces sp. were found similarly to be at pH's around 5.4 and 6.0 while pH 4.8 was optimum for the activities of CMCase and Avicelase from A.niger, indicating that A.niger in acidic media would yield an enzyme of high activity.

• Journal of Microbiology and Biotechnology
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• 제13권1호
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• pp.146-148
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• 2003

Effects of propylamine on conidial head formation and growth of Aspergillus niger and Aspergillus oryzae were analyzed. Propylamine inhibited conidial head formation in these two fungi, and the inhibitory effect of propylamine was not suppressed by the addition of potassium chloride at high concentration, which promotes conidial head formation in A. nidulans and A. oryzae. Propylamine also inhibited the growth of A. niger and A. oryzae by $41\%\;and\;32\%$, respectively, when the concentration of propylamine was $2\%$.

• Journal of Microbiology and Biotechnology
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• 제10권1호
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• pp.51-55
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• 2000

Surface culture fermentation using Aspergillus niger was studied for gluconic and citric acid production at different C/N ratios. A culture of A. niger was found to produce either gluconic acid alone, a mixture of gluconic and citric acid, or citric acid alone depending on the level of nitrogen in the medium (4 to 18mM). Glucose oxidase from the mycelial mat was also analyzed at different levels of nitrogen in the media. By choosing the level of nitrogen in the medium at the start of fermentation, it is possible to produce either of the two acids as the dominant product or the two together as a mixture.

• Food Science and Biotechnology
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• 제16권2호
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• pp.243-248
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• 2007

A biochemical characterization of the tannases from Paecilomyces variotii (produced at Unicamp), Aspergillus niger (purchased from Industrial Kerry Bio-Science) and A. niger cnpat 001 (purchased from Embrapa Agroindustrial Tropical-Brazil) was carried out. P variotii is a new strain obtained from the screening of 500 fungi that were tested for their production of tannase. The biochemical properties of this new tannase from P variotii were determined and compared with those of two other tannase preparations. The tannase produced from P. variotii showed optimum activity at pH 6.5. The functional temperature range of the tannases was from $20-70^{\circ}C$, with optima at $70^{\circ}C$ for P. variotii and at $60^{\circ}C$ for the commercially obtained tannase, whereas A. niger cnpat 001 showed optimum activity at $40^{\circ}C$. The effects of 1 mM preparations of cations and anions, inhibitors, surfactants, and chelators on the tannase activity from P. variotii were also studied.

• Mycobiology
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• 제29권3호
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• pp.132-134
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• 2001

Since it is known that Agrobacterium tumefaciens, which has long been used to transform plants, can transfer the T-DNA to yeast Saccharomyces cerevisiae during tumourigenesis, a variety of fungi were subjected to transformation to improve their transformation frequency. In this study, I report the A. tumefaciens-mediated transformation of filamentous fungus Aspergillus niger. Transfer of the binary vector pBIN9-Hg, containing the bacterial hygromycin B phosphotransferase gene under the control of the Aspergillus nidulans trpC promoter and terminator as a selectable marker, led to the selection of $50{\sim}100$ hygromycin B-resistant transformants per $1{\times}10^7$ conidia of A. niger. This efficiency is improved $10{\sim}20$ fold more than reported elsewhere. In order to avoid the difficulties in selection transformant from the over-growing non-transformant, I used top agar containing 900 ${\mu}g/ml$ of hygromycin. Genomic PCR and Southern analysis showed that all transformants contained single T-DNA insert per fungal genome. This technique offers an easier and more efficient method than that of using protoplast.

• Journal of Microbiology and Biotechnology
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• 제30권2호
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• pp.196-205
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• 2020

In this study, the acidic lipase from Aspergillus niger (ANL) was homologously expressed in A. niger. The expression of ANL was significantly improved by the expression of the native ANL with the introns, the addition of the Kozak sequence and the optimization of the signal sequences. When the cDNA sequence of ANL fused with the glaA signal was expressed under the gpdA promoter in A. niger, no lipase activity could be detected. We then tried to improve the expression by using the full-length ANL gene containing three introns, and the lipase activity in the supernatant reached 75.80 U/ml, probably as a result of a more stable mRNA structure. The expression was further improved to 100.60 U/ml by introducing a Kozak sequence around the start codon due to a higher translation efficiency. Finally, the effects of three signal sequences including the cbhI signal, the ANL signal and the glaA signal on the lipase expression were evaluated. The transformant with the cbhI signal showed the highest lipase activity (314.67 U/ml), which was 1.90-fold and 3.13-fold higher than those with the ANL signal and the glaA signal, respectively. The acidic lipase was characterized and its highest activity was detected at pH 3.0 and a temperature of 45℃. These results provided promising strategies for the production of the acidic lipase from A. niger.

• 생명과학회지
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• 제13권1호
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• pp.105-109
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• 2003

Aspergillus niger #33로부터 RT-PCR 및 PCR방법을 이용하여 epoxide hydrolase (EH)유전자를 클로닝 하고 염기서 열을 분석한 결과 A. niger LCP521 유래의 EB와 85%수준의 유사성을 가지고 있었다. 클로닝된 EH 유전자를 Sacrharomyces cerevisiae에 형질전환 시킨 후 galactose를 inducer로 사용하여 발현시켰다. 유전자 재조합 S. cerevisiae는 라세믹 phenyl oxirane 기질에 대하여 입체선택적 가수분해능이 있음을 확인할 수 있었으며, 이러한 유전자 재조합 EH는 광학활성 에폭사이드 제조를 위한 생촉매로 응용될 수 있을 것이다.

• Archives of Pharmacal Research
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• 제18권6호
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• pp.458-461
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• 1995

+To prove uniqueness of allylic alcohol formation from ${\alpha},{\beta}-unsaturated$ ketones by mammal enzymes, a metabolic pattern of dehydroparadols, non-pungent synthetic analogs of shogaol by Aspergillus niger was examined. Two biotransformation products of a dehydroparadol, 1-(4hydroxy-3-methoxyphenyl)-non-1-en-3-one were accumulated in the culture broth of A. niger. They were characterized as 1-(4-hydroxy-3-methoxyphenyl)-non-1-en-9-01-3-one and 1-(4-hydroxy-3-methoxyphenyl)-nonan-9-o1-3-one by UV, NMR and mass spectroscopic analyses. Accumulation of allylic alcohol metabolites was not observed.

• Journal of Microbiology and Biotechnology
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• 제27권8호
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• pp.1379-1385
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• 2017

The content of taxol in the bark of yews is very low, and this is not affordable from the environmental point of view. Thus, it is a necessity to look for alternative sources of taxol production to solve its supply. Currently, a large portion of the taxol in the market comes from chemical semi-synthesis, but the semi-synthetic precursors such as baccatin III and 10-deacetyl-baccatin III are extracted from needles and twigs of yew trees. Taxol-producing fungi as a renewable resource is a very promising way to increase the scale of taxol production. Our group has obtained a taxol-producing endophytic fungus, Aspergillus niger subsp. taxi HD86-9, to examine if A. niger can produce the taxanes. Six compounds from the fermentation broth of strain HD86-9 were isolated and identified by $^1H$ NMR, $^{13}C$ NMR, and ESI-MS. The results showed that the six compounds included four taxane diterpenoids (taxol, cephalomannine, baccatin III, and 10-deacetyl-baccatin III) and two non-taxane compounds (${\beta}-sitosterol$ and flavonoid isovitexin). The study verified that the taxanes can be produced by the A. niger, which is very important to taxol production via chemical semi-synthesis. Additionally, the finding is potentially very significant to solve the taxol semi-synthetic precursors extracted from needles and twigs of yew trees, and the precursor production can be easily increased through the culture condition optimization, genetic breeding, and metabolic engineering of the A. niger.