The Japonica cultivar 'Janganbyeo' was cultivated and about half of paddy field was totally lodged at 30 days after heading. Both lodged paddy and unlodged paddy were harvested at intervals. The lodged paddy and brown rice did not show any signs of viviparity until the $4^{th}$ day of lodging. The activities of $\alpha$-amylase, diastase, and lipase of lodged paddy and brown rice began to increase on the $2^{nd}$ day of lodging and then increased slowly afterwards. The activities of protease and phytase of lodged paddy and brown rice reached the maximum values on the $2^{nd}$ day and then decreased until the final day of lodging. In contrast, unlodged paddy and brown rice showed relatively small increase in viviparity and in activities of $\alpha$-amylase, diastase, protease, lipase, and phytase. Especially, activities of diastase and lipase were observed even in unlodged paddy and brown rice.
Objective: This study was conducted to determine catalytic properties of wheat phytase with exopolyphosphatase activity toward medium-chain and long-chain inorganic polyphosphate (polyP) substrates for comparative purpose. Methods: Exopolyphosphatase assay of wheat phytase toward polyP75 (medium-chain polyP with average 75 phosphate residues) and polyP1150 (long-chain polyP with average 1150 phosphate residues) was performed at pH 5.2 and pH 7.5. Its activity toward these substrates was investigated in the presence of Mg2+, Ni2+, Co2+, Mn2+, or ethylenediaminetetraacetic acid (EDTA). Michaelis constant (Km) and maximum reaction velocity (Vmax) were determined from Lineweaver-Burk plot with polyP75 or polyP1150. Monophosphate esterase activity toward p-nitrophenyl phosphate (pNPP) was assayed in the presence of polyP75 or polyP1150. Results: Wheat phytase dephosphorylated polyP75 and polyP1150 at pH 7.5 more effectively than that at pH 5.2. Its exopolyphosphatase activity toward polyP75 at pH 5.2 was 1.4-fold higher than that toward polyP1150 whereas its activity toward polyP75 at pH 7.5 was 1.4-fold lower than that toward polyP1150. Regarding enzyme kinetics, Km for polyP75 was 1.4-fold lower than that for polyP1150 while Vmax for polyP1150 was 2-fold higher than that for polyP75. The presence of Mg2+, Ni2+, Co2+, Mn2+, or EDTA (1 or 5 mM) exhibited no inhibitory effect on its activity toward polyP75. Its activity toward polyP1150 was inhibited by 1 mM of Ni2+ or Co2+ and 5 mM of Ni2+, Co2+, or Mg2+. Ni2+ inhibited its activity toward polyP1150 the most strongly among tested additives. Both polyP75 and polyP1150 inhibited the monophosphate esterase activity of wheat phytase toward pNPP in a dose-dependent manner. Conclusion: Wheat phytase with an unexpected exopolyphosphatase activity has potential as a therapeutic tool and a next-generational feed additive for controlling long-chain polyP-induced inappropriate inflammation from Campylobacter jejuni and Salmonella typhimurium infection in public health and animal husbandry.
A gene coding $endo-{\beta}$,-1, 4 glucanase from Actinomyces sp. KNG40 and phytase from Hansenula polymorpha were cloned into Esherichia coli JM101 by using E. coli/Lactobacillus shuttle vector pNZ3004 and pNZ123. The plasmid p3PS(1-4) and p123(1-4) have slpA promoter and slpA signal sequence. So, I constructed expression vectors, p3PS(1-4)Endo, phy and p123(1-4)Endo, phy. These constructed vector was transformed in target host Lactobacillus gasseri and reutri. These transformed host expressed endoglucanase and phytase as extracellular fraction. In the enzyme activity of the same vector, host L, gasseri was higher activity than L. reuteri. This indicates that L. gasseri recongnize promoter and signal sequence very well.
Fandrejewski, H.;Raj, S.;Weremko, D.;Zebrowska, T.;Han, In K.;Kim, J.H.;Cho, W.T.
Asian-Australasian Journal of Animal Sciences
/
제10권6호
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pp.665-670
/
1997
The digestibility (apparent) of P and other nutrients from the RSM, SBM and 2 batches of maize, wheat and barley was investigated in two experiments with 24 castrated male growing pigs. The effect of supplemental microbial phytase (1,000 U/kg) was also evaluated. The diets contained 25% RSM (Exp. 1) or 40% SBM (Exp. 2) and had no inorganic P. In each period of digestive trial, after 9 days of adaptation, faeces were collected for 5 days. The digestibility of P contained in the RSM and SBM was calculated by difference method. The P digestibility in maize, wheat, barley was on average 20, 34 and 36%, respectively. The digestibility of P in the RSM and SBM estimated from maize-based diets were 19 and 24 %, respectively. Kind of cereal grain had significant (p < 0.05) influence on the digestibility of P which was lower in the diets based on maize than wheat or barley. The digestibility of P significantly increased with the supplemental microbial phytase (on an average of 17%). Moreover, inclusion of enzyme into the diets positively affected digestibility of other nutrients, namely the protein and organic matter.
This study was conducted to investigate the effects of dietary supplementation with multiple enzymes composed of phytase plus carbohydrolases (ROVABIO$^{(R)}$ Max, RM) on growth performance, nutritional availability and intestinal viscosity in broiler chicks. A total of one thousand, one-day-old male broiler chicks were randomly allotted into treatment groups that received one of five experimental diets for 32 days. Each group consisted of 40 birds and all experiments included five replicates. The dietary treatments included PC (a positive control diet), NC1 (65 kcal/kg, 0.15% and 0.10% less ME, available phosphorus and calcium levels, respectively, than the PC diet), NC2 (85 kcal/kg, 0.20% and 0.10% less ME, available phosphorus and calcium levels, respectively, than the PC diet), NC1+RM (NC1 plus ROVABIO$^{(R)}$ Max) and NC2+RM (NC2 plus ROVABIO$^{(R)}$ Max). The average body weights, daily body weight gains and feed conversion rates of the chicks fed a diet containing RM improved significantly or tended to improve. The treatments also had no effect on the carcass characteristics or blood parameters, but the viscosity of the intestinal contents of the chicks fed the diet containing RM was significantly lower than that of chicks in the NC without RM groups. Additionally, chicks fed the dietary RM showed increased breaking strength and ash content of the tibia when compared to chicks that received the non-RM diets. Taken together, the results of the present study indicated that the addition of multiple enzymes consisting of phytase plus NSP enzymes improved the growth performance and mineral status of the tibia in broiler chickens fed corn-wheat-soybean meal-based diets with reduced levels of nutrients. Further, these findings suggest that the improved animal performance is associated with reduced intestinal viscosity by the dietary enzyme complex.
Ilyas, A.;Hirabayasi, M.;Matsui, T.;Yano, H.;Yano, F.;Kikishima, T.;Takebe, M.;Hayakawa, K.
Asian-Australasian Journal of Animal Sciences
/
제8권2호
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pp.135-138
/
1995
Soybean meal was fermented by Aspergillus usami in order to reduce phytate content. Aflatoxin B1 was not detected in the fermented soybean meal. The contents of crude protein, crude fiber, ether extract and crude ash were slightly increased following fermentation with a concomitant reduction in nitrogen free extract. Though the fermentation partly degraded proteins in the soybean meal, there was small difference in amino acid composition between the soybean meal and the fermented soybean meal. The results showed that the fermentation did not affect nutritional value of protein in soybean meal. Approximately 55% of phosphorus extracted by trichloroacetic acid was inositol hexaphosphate (phytate) in the soybean meal. The content of inositol tetra to hexaphosphates was not detected in the fermented soybean meal. These results indicated that the fermentation almost completely eliminated phytate in soybean meal. Phytase activity was not detected in the unfermented soybean meal. However, the enzyme activity in the fermented soybean meal was 167.7 U/g. When the fermented soybean meal in supplemented in formula feeds, phytase in the fermented soybean meal might partly degrade the phytate in other ingredients in the digestive tract. The fermented soybean meal is possibly used as a phytate-free protein source of feed, which contains high available phosphorus.
Huang, Chung Ming;Chuang, Wen Yang;Lin, Wei Chih;Lin, Li Jen;Chang, Sheng Chang;Lee, Tzu Tai
Animal Bioscience
/
제34권3_spc호
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pp.371-384
/
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.
Kim, Young-Ok;Park, In-Suk;Nam, Bo-Hye;Kong, Hee-Jeong;Kim, Woo-Jin;Lee, Sang-Jun;Kim, Kyung-Kil
Fisheries and Aquatic Sciences
/
제13권1호
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pp.49-55
/
2010
A bacterial strain with phytase and glucose-1-phosphatase activity was isolated from seawater. The colony was identified as an Enterobacter cloacae strain and named E. cloacae B11. A gene, agpEnB11, coding for an intracellular acid glucose phosphatase was cloned from the strain and sequenced. It comprised 1,242 nucleotides and encoded a polypeptide of 413 amino acids. Recombinant glucose-1-phosphatase (AgpEn) was overexpressed in Escherichia coli and purified using Ni-NTA column under native conditions. Purified protein displayed a single band of 47 kDa on SDS-PAGE. AgpEn hydrolyzed a wide variety of phosphorylated compounds, with high activity for glucose-1-phosphate and glucose-6-phosphate. Optimum pH and temperature for enzyme activity were pH 5.0 and $50^{\circ}C$, respectively. Enzyme activity was stimulated by $Ca^{2+}$ and $Co^{2+}$, and inhibited by $Cu^{2+}$.
Xuan, Z.N.;Kim, J.D.;Lee, J.H.;Han, Y.K.;Park, K.M.;Han, In K.
Asian-Australasian Journal of Animal Sciences
/
제14권2호
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pp.231-236
/
2001
This study was conducted to investigate the effect of supplemented enzyme complex on growth performance and nutrient digestibility in pigs weaned at 14 days of age. Eighty pigs ($4.02{\pm}0.11kg$ of average body weight) were allotted in a completely randomized block design. Treatments were as follows: 1) control (negative), 2) control (positive, $Kemzyme^{(R)}$), 3) 0.1%, 4) 0.2% and 5) 0.3% of newly developed enzyme complex. Each treatment has 4 replicates with 4 pigs per replicate. During phase I period (d 0 to 14), ADG and ADFI were numerically higher in pigs fed diets supplemented enzyme complex regardless of their inclusion levels compared to pigs fed control (negative) diet. Feed/gain (F/G) was also better in pigs fed enzyme complex diet than that of pigs fed control (negative) diet. In addition, with increasing the inclusion level of enzyme complex, ADG and ADFI were improved. However, there was no significant difference between treatment in all growth parameters. During phase II period (d 15 to 28), ADG, ADFI and F/G showed the same tendency as in phase I period. For overall period (d 0 to 28) ADG was highest in pigs fed diet included 0.2% enzyme complex in all treatments but not significantly different. During phase I period, the digestibilities of all nutrients did not showed any significant difference between treatments. However, pigs fed diet contained enzyme complex and positive control diet (Kemzyme) showed numerically higher nutrient digestibilities in all nutrients than pigs fed negative control diet. During phase II period, data were consistent with those observed in phase I period. Especially, the digestibility of phosphorus was significantly higher in pigs fed diets contained enzyme complex including phytase than pigs fed control (negative and positive) diets (p<0.05). For overall experimental period, fecal or ileal amino acid digestibility were not affected by dietary treatment. Enzyme complex newly developed and used in this study can be possibly recommended as a growth promoter when supplemented in diet for early weaned piglets.
Three trials were conducted to determine the available energy of different wheat screening varieties collected from different locations of Khorasan in Iran. In experiment 1, chemical composition and the nitrogen corrected true metabolisable energy (TMEn) were evaluated. A precision-fed rooster assay was used, in which, each wheat screening sample was tube fed to adult roosters, and the excreta were collected for 48-h. In Exp. 2 and 3, five and two wheat screening verities-based diets with or without xylanase and phytase were fed to 16-day old battery reared chicks respectively, and total feed consumption and excreta were measured during next three days. The variable nature of wheat screening varieties led to significant differences in mean TMEn values (p<0.01). The TMEn values of samples determined with adult roosters varied by ${\pm}5.03%$ of the mean value ($3,097.65{\pm}49.32\;kcal/kg$) and ranged from 2,734.90 to 3,245.12 kcal/kg. There was a significant correlation (p<0.05) between crude fiber (CF), neutral detergent fiber (NDF), and acid detergent fiber (ADF) with TMEn, and the greatest correlation coefficient was observed between NDF and TMEn (r = -0.947; p<0.001). The optimal equation in terms of $R^2$ from using a single chemical analysis was obtained with NDF: TMEn = 4,152.09-27.80 NDF ($R^2$ = 0.90, p<0.0001), and the TME prediction equation was improved by the addition of the crude protein (CP) and ASH content to sequential analysis: TMEn = 3,656.97-28.65 NDF+32.54 CP+38.70 ASH ($R^2$ = 0.98, p<0.0001). The average AMEn values of 5 and 2 wheat screening varieties determined with young broiler chickens were $2,968.41{\pm}25.70\;kcal/kg$ and $2,976.38{\pm}8.34\;kcal/kg$ in Exp. 2 and Exp. 3, respectively. Addition of xylanase and phytase to wheat screenings resulted in significant (p<0.01) improvement in AMEn by 4.21 and 2.92%, respectively.
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