• Asian-Australasian Journal of Animal Sciences
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• v.27 no.9
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• pp.1285-1292
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• 2014

An in vitro experiment was conducted to evaluate the effects of Aspergillus oryzae culture (AOC) and 2-hydroxy-4-(methylthio)-butanoic acid (HMB) on rumen fermentation and microbial populations between different roughage sources. Two roughage sources (Chinese wild rye [CWR] vs corn silage [CS]) were assigned in a $2{\times}3$ factorial arrangement with HMB (0 or 15 mg) and AOC (0, 3, or 6 mg). Gas production (GP), microbial protein (MCP) and total volatile fatty acid (VFA) were increased in response to addition of HMB and AOC (p<0.01) for the two roughages. The HMB and AOC showed inconsistent effects on ammonia-N with different substrates. For CWR, neither HMB nor AOC had significant effect on molar proportion of individual VFA. For CS, acetate was increased (p = 0.02) and butyrate was decreased (p<0.01) by adding HMB and AOC. Increase of propionate was only occurred with AOC (p<0.01). Populations of protozoa ($p{\leq}0.03$) and fungi ($p{\leq}0.02$) of CWR were differently influenced by HMB and AOC. Percentages of F. succinogenes, R. albus, and R. flavefaciens (p<0.01) increased when AOC was added to CWR. For CS, HMB decreased the protozoa population (p = 0.01) and increased the populations of F. succinogenes and R. albus ($p{\leq}0.03$). Populations of fungi, F. succinogenes (p = 0.02) and R. flavefacien (p = 0.03) were increased by adding AOC. The HMB${\times}$AOC interactions were noted in MCP, fungi and R. flavefacien for CWR and GP, ammonia-N, MCP, total VFA, propionate, acetate/propionate (A/P) and R. albus for CS. It is inferred that addition of HMB and AOC could influence rumen fermentation of forages by increasing the number of rumen microbes.

• Asian-Australasian Journal of Animal Sciences
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• v.12 no.3
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• pp.417-421
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• 1999

This experiment examined the effects of feeding Aspergillus oryzae (AO) culture to laying hens, on fecal microbial populations, fecal pH and moisture content, egg quality, and metabolizabilities of several nutrients. Sixteen commercial 38-wk-old laying hens were randomly allotted to four diets: control; with 0.15% locally produced AO culture; with 0.3% locally produced AO culture, and; or with 0.3% imported AO. Each treatment consisted of four replicates (cages) containing one bird per cage according to a completely randomized design. After 4 wk, AO were recovered in the feces of birds fed the AO diets, indicating that AO might pass through the fore-gut alive and become active in the hind gut. The number of Lactobacillus spp. in feces was higher in all treated groups than that of the control, indicating that AO would provide a beneficial environment for the Lactobacillus spp. to proliferate in the intestine. The number of fecal E. coli was significantly reduced by the addition of AO. A similar trend was also found for aerobic bacteria. Although not significant, fecal moisture contents tended to be reduced by the addition of AO. Fecal pH was not significantly different among the treatments. The addition of AO did not affect the various economic traits of eggs. Metabolizabilities of gross energy and dry matter measured during the 5th wk were increased by the AO supplementation. It appears that AO culture alone could be used as a probiotic supplement for layers.

• Preventive Nutrition and Food Science
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• v.20 no.1
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• pp.29-37
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• 2015

Most of the wheat germ in cereal grains is removed during the milling process. Various physiological effects have been reported for bioactive substances in wheat germ such as phenolic acids and flavonoids. In this study, the antioxidant and anti-adipogenic effects of ethanol extracts from wheat germ (WGE) and wheat germ fermented with Aspergillus oryzae (F-WGE) were investigated in HepG2 and 3T3-L1 cells. The anti-oxidant activity of F-WGE was demonstrated by a dose-dependent increase in the enhanced scavenging capacity of hydroxyl radicals and $Cu^{2+}$-chelating activity compared to WGE. WGE and F-WGE treatment at doses between 10 and $400{\mu}g/mL$ did not affect the viability of HepG2 and 3T3-L1 cells. Intracellular ROS levels from $Cu^{2+}$-induced oxidative stress were significantly decreased by F-WGE treatment in HepG2 cells compared to WGE. Lipid accumulation was increased in 3T3-L1 adipocytes by $100{\mu}M$ $Fe^{2+}$ treatment, but the accumulation was strongly inhibited by $100{\mu}g/mL$ of WGE and F-WGE treatment. These results suggest that changes in bioactive substances during the fermentation of wheat germ can potentiate scavenging activities against transition metal-induced oxidative stress and lipid accumulation in 3T3-L1 adipocytes. Therefore, we propose that F-WGE is a novel food materials and provided scientific evidences for its efficacy in the development of functional foods.

• Asian-Australasian Journal of Animal Sciences
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• v.13 no.8
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• pp.1076-1083
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• 2000

The aim of this study was to evaluate the effect of Aspergillus oryzae fermentation extract (AFE) on in situ degradation of the various concentrates, forages and by-products in Taiwan. The in situ trial was conducted to determine the effect of AFE on the rate of ruminal degradation of dry matter (DM), organic matter (OM), neutral detergent fiber (NDF), and acid detergent fiber (ADF) of the various local available feedstuff commonly used for dairy cattle. Two ruminal fistulated cows were arranged into a two by two switchback trial. Two dietary treatments were control without AFE inclusion diet and diet with 3 g of AFE (Amaferm) added daily into the total mixed ration (TMR). Results showed that effect of AFE inclusion on the ruminal degradability of concentrates vary; soybean meal is the most responsive feedstuff, corn is the next, whereas full-fat soybean did not response the AFE inclusion at all. The inclusion of AFE significantly depressed most of the nutrient degradation of the concentrates of soybean meal in the first 12-hour in situ incubation. The effect declined in the next 12 hours. Rapeseed meal showed a different trend of response: addition of AFE improved its NDF degradation. The inclusions of AFE significantly improved ADF degradation of roughage after 24 or 48 hours of incubation. However, corn silage and peanut-vines showed a different trend. Effects of AFE inclusion on the by-products degradability were inconsistent. Most of nutrients in rice distillers grain and some in beancurd pomace did show increased degradation by the AFE inclusion.

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.1
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• pp.53-62
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• 2004

The aim of this study was to predict the energy value and dynamic degradation of roughage in Taiwan using the $Daisy^{(R)}$. in vitro fermentation method to provide information on one of the very important nutrients for ration formulation. The second objective was to study the effects of Aspergillus oryzae (AFE) inclusion on nutrient utilization. Three ruminal fistulated dry dairy cows were used for rumen fluid and fifteen conventional forages used in dairy cattle were collected around this island. The degradability of these feedstuffs with and without AFE ($Amaferm^{(R)}$.) treatment was measured using the $Daisy^{(R)}$. in vitro method. The roughage energy values, including TDN and NEL, were calculated according to Robinson (2000). Results from the 30 h in vitro neutral detergent fiber (NDF) degradability and predicted energy evaluations showed that alfalfa (among the forages) contained the highest degradability and energy values, Bermuda straw having the lowest. Peanut vines and corn silage contained higher energy values and the lowest value found in Pangola and Napier grasses among the locally produced forages. Pangola and Napier grasses had lower values than most imported forages except Bermuda straw. Among the by-products, wheat middling contained the highest NDF degradability, while rice bran contained the richest energy value due to its high oil content. From the dynamic dry matter (DM), organic matter (OM), acid detergent fiber (ADF) and neutral detergent fiber (NDF) degradation, corn silage contained the highest effective degradation among the local forages; wheat middling (among the by-products) degraded the fastest in DM, OM, ADF and NDF and showed the highest effective degradability. AFE inclusion was inconsistent among the forages. Alfalfa hay showed significantly increased 30 h NDF degradability and energy values, Pangola hay, Napier grass and brewer's grains showed decreased degradability and energy values. AFE inclusion increased the DM, OM and NDF degradation rate in most forage, but only increased the DM degradation rate in sorghum distiller's grains, the OM degradation rate in bean curd pomace and the NDF and ADF degradation rates in soy pomace (among the by-products).

• Korean Journal of Fisheries and Aquatic Sciences
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• v.32 no.6
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• pp.725-729
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• 1999

In order to utilize large size anchor effectively, fermented and powdered anchovy seasoning for extractives was manufactured, Fermented anchovy saesoning was fabricated by adding $10\%$ koji of Aspergillus oryzae and mixing with $5\%$ Laminalia. The optimum fermentation temperature, humidity and time for manufacture of anchovy saesoning were $40^{\circ}C$, RH $80\%$ and 48 hrs, respectively. The amount of total free amino acids in anchovy seasoning with $5\%$ Lansinaria was 6,486.9 mg/100 g, while that of commercial product was 444.4 mg/100 g. The principal taste compounds in anchovy seasoning material were IMP and amino acids such as leucine, glutamic acid, histidine, alanine and valine. Extractive nitrogen and organoleptic quality of the extractives in anchovy seasoning packed in tea bag with air permeability, 100 $m^3/m^2/min$, were better than those of commercial product.

• Journal of Microbiology and Biotechnology
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• v.26 no.2
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• pp.347-355
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• 2016

An exo-β-D-glucosaminidase (AorCsxA) from Aspergillus oryzae FL402 was heterologously expressed and purified. The deduced amino acid sequence indicated that AorCsxA belonged to glycoside hydrolase family 2. AorCsxA digested colloid chitosan into glucosamine but not into chitosan oligosaccharides, demonstrating exo-β-D-glucosaminidase (CsxA) activity. AorCsxA exhibited optimal activity at pH 5.5 and 50℃; however, the enzyme expressed in Pichia pastoris (PpAorCsxA) showed much stronger thermostability at 50℃ than that expressed in Escherichia coli (EcAorCsxA), which may be related to glycosylation. AorCsxA activity was inhibited by EDTA and most of the tested metal ions. A single amino acid mutation (F769W) in AorCsxA significantly enhanced the specific activity and hydrolysis velocity as revealed by comparison of V_max and k_cat values with those of the wild-type enzyme. The three-dimensional structure suggested the tightened pocket at the active site of F769W enabled efficient substrate binding. The AorCsxA gene was heterologously expressed in P. pastoris, and one transformant was found to produce 222 U/ml activity during the high-cell-density fermentation. This AorCsxA-overexpressing P. pastoris strain is feasible for large-scale production of AorCsxA.

• Animal Bioscience
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• v.34 no.3_spc
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• pp.371-384
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• 2021

Objective: Wheat bran (WB) was co-fermented with Aspergillus oryzae and phytase (Phy) to determine whether co-fermentation improve WB phosphorus and fiber utilization in Isa-brown layers. Methods: A total of 112 Isa brown layer were randomly divided into 7 treatments with 8 replicates per a treatment and 2 hens per a replicate. The treatments included basal diet (control), basal diet supplemented with 250 unit/kg Phy (control+Phy), diet with 10% WB (10% WB), diet with 5% WB and 250 unit/kg Phy (5% WB+Phy) diet with 10% WB and 250 unit/kg Phy (10% WB+Phy), diet with 5% fermented WB supplemented with molasses and phy (PCFWH) and 125 unit/kg Phy (5% PCFWH), and diet with 10% PCFWH (10% PCFWH). The intestinal microbial population, intestinal morphology, serum antioxidant enzyme activities, and excreta phosphorus content were assessed. Results: In PCFWH, spore counts, protease activity, xylanase activity, and ferulic acid were 8.50 log/g dry matter (DM), 190 unit/g DM, 120 unit/g DM, and 127 ㎍/g, respectively. Xylobiose and xylotriose were released in PCFWH, while they were not detectable in WB. Antioxidant capacity was also enhanced in PCFWH compared to WB. The 10% WB+Phy and 10% PCFWH groups produced higher egg mass, but hens fed 5% WB+Phy had the lowest amount of feed intake. Eggs from 10% PCFWH had better eggshell weight, eggshell strength, and eggshell thickness. Birds fed with 10% PCFWH also had higher serum superoxide dismutase and catalase activities. Compare to control, 10% PCFWH significantly reduced excreta phosphorus content. Conclusion: Diet inclusion of 10% PCFWH improved egg quality, antioxidant status, and excreta phosphorus content of laying hens.

• Journal of the Korean Society of Food Science and Nutrition
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• v.37 no.2
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• pp.141-147
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• 2008

This study investigated the isoflavone contents and physiological properties of non-fermented soybean (NF) and the fermented soybeans prepared with Asp. oryzae (AO) and B. natto (BN). The total isoflavone contents (daidzin, genistin, daidzein and genistein) of NF, AO and BN were 81.8 mg/100 g, 130.7 mg/100 g and 139.5 mg/100 g, respectively. Especially, the total phenol contents of NF, AO and BN were 2.1%, 4.3% and 7.6%, and the total flavonoid contents were 1.3%, 1.6% and 2.7%, respectively. The nitrite-scavenging abilities of NF, AO and BN were 34.4%, 55.2% and 92.5%, respectively. Antimicrobial activity of BN was shown to be the strongest to Bacillus cereus, Staphylococcus aureus and Escherichia coli. The SOD-like activity was the strongest in AO, whereas the electron donating ability was the strongest in BN. Antioxidant activity of AO at concentration of 0.02% was stronger than BN or NF.

• Korean Journal of Microbiology
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• v.32 no.1
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• pp.84-90
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• 1994

Kinetic parameters of various substrates and inhibitors were measured to elucidate the binding requirements of the active site of intracellular adenosine deaminase (ADA) in Aspergillus oryzae. 3'-Deoxyadenosine was the best substrate according to the value of relative kcat/$K_m$. Purine riboside was found to be the strongest inhibitor with the $K_i$ value of $3.7{\times}10^{-5}$M. Adenine acted neither as a substrate nor as an inhibitor, suggesting the presence of ribose at N-9 of adenosine was crucial to binding. ADA also catalyzed the dechlorination of 6-chloropurine riboside, generating inosine and chloride ions. Substrate specificity of 6-chloropurine riboside was 0.86% of adenosine. Purine riboside, a competitive inhibitor of ADA, inhibit the dechlorination with similar $K_i$ value, suggesting that the same binding site was involved in deamination and dechlorination. Among the sulfhydryl group reagents, mercurials, pchloromercuribenzoate (PCMB), mersalyl acid and $HgCl_2$ inactivated the enzyme. Mersalyl acid-inactivated ADA was reactivated by thiol reagents, but PCMB-inactivated enzyme was not. When ADA was treated with the mercurial reagents, the inhibition constants and inhibition patterns were determined. Each inhibition was competitive with substrate. The $K_i$ values of these mercurial reagents were lower in 10 mM phosphate buffer than in 100 mM phosphate buffer, showing phosphate dependency.