• KSBB Journal
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• v.19 no.1
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• pp.42-49
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• 2004

A gene coding for epoxide hydrolase (EH) of Aspergillus niger LK, a fungus possessing the enantioselective hydrolysis activity for racemic epoxides, was characterized by phylogenetic analysis. The deduced protein of A. niger LK epoxide hydrolase shares significant sequence similarity with several bacterial EHs and mammalian microsomal EHs (mEH) and belongs to the a/${\beta}$ hydrolase fold family. EH from A. niger LK had 90.6% identity with 3D crystal structure of lqo7 in Protein Data Bank. Sequence comparison with other source EHs suggested that Asp$\^$l92/, Asp$\^$374/ and His$\^$374/ constituted the catalytic triad. Based on the multiple sequence comparison of the functional and structural domain sequence, the phylogenetic tree between relevant epoxide hydrolases from various species were reconstructed by using Neighbor-Joining method. Genetic distances were so far as 1.841-2.682 but characteristic oxyanion hole and catalytic triad were highly conserved, which means they have diverged from a common ancestor.

• Applied Chemistry for Engineering
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• v.17 no.5
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• pp.557-560
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• 2006

The epoxide hydrolase (EH) of Aspergillus niger LK was expressed to high levels in Escherichia coli based on codon usage. E. coli, Rosetta (DE3)PLysS, containing a large number of tRNAs for rare-codons was employed as a host strain. The recombinant E. coli expressing A. niger EH showed an enhanced enantioselective hydrolysis activity toward racemic styrene oxide. Enantiopure (S)-styrene oxide with a high enantiopurity of 99% ee was obtained from racemic substrates.

• Journal of Life Science
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• v.28 no.8
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• pp.969-976
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• 2018

In this study, transglucosidase (TG), an enzyme produced by Aspergillus niger, synthesized isomaltooligosaccharide from ${\alpha}-(1{\rightarrow}4)$ linked substrates. The highest TG-producing A. niger KCTC6913 was selected from six kinds of species, and optimized TG producing conditions were established. Five different carbon sources (potato starch, sweet potato starch, corn starch, wheat starch, and dextrin) and three different nitrogen sources (yeast extract, malt extract, and beef extract) were tested to establish the carbon and nitrogen sources favorable for TG production. Measurements of TG activity after an initial culture at pH 5.0 for 15 days revealed that potato starch and yeast extract, which are basic culture media, resulted in the highest TG activity. In addition, A. niger KCTC6913 increased TG production under aerobic conditions and a controlled carbon/nitrogen ratio. In conclusion, to evaluate TG activity in the established optimal medium, it is confirmed that the basal and potato dextrose broth medium were used as a control, and the highest TG production was measured, which was highlighted in the established optimal medium.

• Microbiology and Biotechnology Letters
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• v.32 no.2
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• pp.154-157
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• 2004

Immobilized cells of Aspergillus niger was employed to produce citric acid by fermentation of milk factory waste water. A. niger ATCC 9142 as a citric acid production strain was cultured for 3 days and was entrapped with Ca-alginate bead about 2.5∼3.5 mm. The optimal pH and temperature were estimated to be 3.0 and $30^{\circ}C$, respectively. Dilution rate for fermentation was calculated to be $0.025 h^{－1}$ . Maximum amount of citric acid was obtained at 4.5 g/$\ell$ with the optimized fermentation condition. The yield of citric acid produced by immobilized A. niger ATCC 9143 was 70.3%. The yield was increased by 20% with immobilized cell, compared to that of the shake flask culture. Hence, the milk factory waste water is worthy to be used for the substrate of citric acid fermentation.

• Journal of Life Science
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• v.6 no.1
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• pp.6-13
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• 1996

The possible use of milk-wastewater as a fermentation media for the production of citric acid by Aspergillus niger has been investigated. The addition of Mn$^{2+}$ , Fe$^{2+}$ and Cu$^{2+}$ to the medium promoted citric acid production while only Mg$^{2+}$ decreased citric acid production. The concentrations of citric acid produced were marked up to 7.2g/l and 16.5g/l in a batch bioreactor by Aspergillus niger ATCC 9142 with 50g/l and 100g/l of reducing sugar concentration in milk-wastewater, respectively. A mathematical model was developed and simulating the predictability of cell growth, citric acid production and substrate consumption rate, and gave good agreement results with experimental data.

• The Korean Journal of Mycology
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• v.15 no.3
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• pp.158-168
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• 1987

Several species of the fungi were isolated from cassava(Manihot esculenta Gruntz) starch which had formed into pellet, those had been stored for a while in southern part of Thailand. The species of Rhizopus, Aspergillus niger, and Aspergillus fumigatus were identified. The experimental results are as follows; Dry weight increases were checked during the static liquid culture with modified Czapek Dox medium to which cassava starch was partly replaced to sugar, Aspergillus niger and Aspergillus fumigatus had grown more than Rizopus species when 6% cassava starch was replaced to sugar and had been cultured for 72 hours. Amounts of mycelial protein of Aspergillus niger were checked, the highest amount was shown in 6% cassava starch involved medium. When nitrogen sources were varied such as ammonium sulfate or urea against sodium nitrate, there was no significant difference in mycelial production. Alpha amylase activity of each fungus isolated here was checked, those of Aspergillus niger have shown the highest peak at 72 hours.

• The Korean Journal of Mycology
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• v.10 no.4
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• pp.193-202
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• 1982

The changes of total carbohydrates and trehalose levels during differentiation of A. niger were studied. Aspergillus niger was cultivated in Czapeck-Dox medium by the method of surface culture and shake culture. Total carbohydrates and trehalose were fractionated and determined by the method of Trevelyan and Harrison (1956). Total carbohydrates and trehalose accumulated in A. niger during sporulation. The influences of nutritional starvation on the levels of cellular carbohydrates in A. niger, which was cultivated in each nitrogen, phosphate and glucose limited Czapeck-Dox medium, were studied. The levels of total carbohydrates and trehalose in A. niger cultivated in nitrogen limited medium increased much than those cultured in full medium. The total carborates and trehalose levels in A. niger cultivated in phosphate limited medium increased, but the increases were less than those cultured in nitrogen limitted medium. In glucose limited medium, any changes of total carbohydrates and trehalose levels were not found. It is considered that the biochemical mechanisms responsible for the changes of total carbohydrates and trehalose levels may be related with differentiation of Aspergillus niger.

• KSBB Journal
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• v.13 no.2
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• pp.214-219
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• 1998

Polymerase chain reaction(PCR) was conducted to obtain the gene encoding glucose oxidase(GOD) from Aspergillus niger(ATCC 2110) and the DNA sequence determined was coincided with published GOD sequence from A. niger. Recombinant transforming vector containing GOD and hygromycin B(hyg.B) resistant gene(hph) was constructed and used for further transformation of A. niger ATCC 2110. Selectivity of hyg.B against A. niger differed depending on which media were used i.e., nutrient-rich media such as potato dextrose agar(PDA) and complete medium(CM) showed only 50% growth inhibition at 400 $\mu$m ml$^-1$ of hyg.B while the minimal media inhibited mycelial growth completely at 200 $\mu$m ml$^-1$ of hyg.B. Twenty to sixty putative transformants were isolated from the hyg.B-containing minimal top agar, transferred successively onto alternating selective and nonselective media for a mitotic stability of hyg.B resistance and, then, single-spored. Among the stable transformants, the transformant(GOD1-6) grown by flask culture showed the considerable increase of extracellular GOD activity, which was estimated to the degree of 50% - 100% comparing to that of wild type. Transformation of tGOD1-6 was resulted from integration of the vectors into heterologous as well as homologous regions of the A. niger genome. Southern blot analysis revealed that there were two independent integrations of vector into fungal genome and one into the GOD gene due to homologous recombination. In addition, GOD activity and sodium gluconate production when tGOD1-6 was fed-batch fermented were enhanced 11 fold and 2.25 fold, respectively, compared to that of the wild type.

• Journal of The Korean Society of Grassland and Forage Science
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• v.30 no.2
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• pp.179-190
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• 2010

The experiment was conducted to observe the effects of dried whole crop barley treated with cellulolytic microorganisms (Aspergillus niger KCCM 60357 and Bacillus licheniformis KCCM 40934) on the chemical composition, in vitro colonic fermentation and whole tract digestibility in swine. Whole crop barley were fermented with no microorganism addition (control), A. niger, B. licheniformis and co-culture of A. niger and B. licheniformis (Mixture) for 3 days at $30^{\circ}C$. In the feed chemical composition, CP contents of whole crop barley treated with A. niger (7.52%) and B. licheniformis (7.77%) were significantly higher than control (6.81%) (p<0.05). The in vitro colonic fermentation of dried whole crop barley fermented with control showed significantly higher $CH_4$ contents than A. niger, B. licheniformis and Mixture at 18h incubation (p<0.05). Dry matter (DM) digestibilities of A. niger (55%) and Mixture (57.42%) treatments were significantly higher than control (43.74%) (p<0.05). Ammonia-N was significantly increased in A. niger, B. licheniformis and Mixture relative to control at 24 hour incubation (p<0.05). Xylanase activities in A. niger, B. licheniformis and Mixture treatments were significantly higher than control at 24 hour incubation (p<0.05). Concentrations of total VFA were significantly increased in B. licheniformis (12.61 mM) at 24hour incubation (p<0.05). In vitro whole tract digestibility was significantly increased in B. licheniformis (49.61%) compared with the control (45.65%) (p<0.05). In conclusion, whole crop barley treated with cellulolytic microorganisms improved whole tract digestibility and colonic fermentation for swine.

• Microbiology and Biotechnology Letters
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• v.24 no.4
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• pp.427-429
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• 1996

We have already cloned an extracellular $\alpha$-glucosidase gene from Aspergillus niger with oligonucleotide probe synthesized on the basis of the peptide sequences determined previously. The DNA sequence revealed an open reading frame of 895 amino acids split by three introns. We are attempting to construct an A. niger strain deficient in the $\alpha$-glucosidase enzyme activity, which would be useful for the glucoamylase production without contamination by the industrially undesirable $\alpha$-glucosidase. For destruction of the $\alpha$-glucosidase gene, we try to make transformations. A cloned partial $\alpha$-glucosidase gene was introduced into Aspergillus niger, and transformants with suppressed $\alpha$-glucosidase activity were isolated. The transformants were cultured on YPD medium which contained Hygromycin B at 30$\circ$C. The activity of $\alpha$-glucosidase of the suppressed transformants was compared to that of wild type activity. As shown by southern-hybridization, we detected that the transformant was a heterocaryon.