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Effects of Amylase and Cellulase Supplementation in Sorghum-based Diets for Finishing Pigs

  • Park, J.S.;Kim, I.H.;Hancock, J.D.;Hines, R.H.;Cobb, C.;Cao, H.;Hong, J.W.;Kwon, O.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.1
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    • pp.70-76
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    • 2003
  • Three experiments were conducted to determine the effects of a sorghum-specific enzyme system, derived from an Aspergillus niger and Bacillus subtilis fermentation extract (carbohydrase activity of 1,650 $\alpha$-amylase units and cellulase activity of 30 fibrinolytic units/mL), on growth performance of finishing pigs. In Exp. 1,192 pigs (average initial BW of 46.1 kg) were fed sorghum-based diets without or with 360 mL of enzyme system per ton of sorghum in a 78 d growth assay. For d 0 to 39, gain/feed was improved (p<0.03) with enzyme supplementation, but ADG was not affected (p>0.15). For d 39 to 78 and overall (d 0 to 78), ADG, gain/feed, and digestibilities of DM and N were not affected (p>0.13) by enzyme supplementation. Backfat thickness, fat-free lean index, and scores for stomach keratinization and ulcers also were not affected (p>0.15) by the dietary treatments. In Exp. 2,168 pigs (average initial BW of 58.4 kg) were fed diets without or with 150, 300, or 450 mL/ton of the same enzyme system used in Exp. 1. Adding as much as 450 mL enzyme system / ton of sorghum did not affect (p>0.15) ADG or gain/feed for d 0 to 29 of the growth assay. However, during d 29 to 63, ADG increased by 11% (linear effect, p<0.02) and gain/feed increased by 10% (linear effect, p<0.06) as enzyme concentration was increased from none to 450 mL/ton of sorghum. For the overall period (d 0 to 63), ADG tended to increase (p<0.08) with enzyme supplementation, but gain/feed and digestibilities of DM and N were not affected (p>0.14). Carcass characteristics (dressing percentage, backfat thickness, and fat free lean index) also were not affected (p>0.20) by addition of the enzyme system. In Exp. 3,176 pigs (average initial BW of 46.7 kg) were fed diets without or with 450, 900, or 1,350 mL/ton of the same enzyme system used in Exp. 1 and 2 in a 71 d growth assay. Adding up to 1,350 mL/ton of enzyme had no effects (p>0.15) on ADG, gain/feed, digestibilities of DM and N, and carcass characteristics (dressing percentage, backfat thickness, and fat-free lean index). In conclusion, finishing pigs fed diets with a sorghum-specific enzyme system showed some positive trends for improved growth performance, but those effects were not large and (or) consistent.

Comparative Efficacy of Different Soy Protein Sources on Growth Performance, Nutrient Digestibility and Intestinal Morphology in Weaned Pigs

  • Yang, Y.X.;Kim, Y.G.;Lohakare, J.D.;Yun, J.H.;Lee, J.K.;Kwon, M.S.;Park, J.I.;Choi, J.Y.;Chae, B.J.
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.5
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    • pp.775-783
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    • 2007
  • To elucidate the efficacy of different soy protein sources on piglet's performance, a total of 280 weaned piglets ($Duroc{\times}Yorkshire{\times}Landrace$, $23{\pm}3$ d of age, $5.86{\pm}0.45$ kg initial BW) were allotted to 5 treatment diets comprising soybean meal (SBM), soy protein concentrate (SPC), Hamlet protein (HP300), fungal (Aspergillus oryzae) fermented soy protein (FSP-A), and fungal plus bacterial (A. oryzae+Bacillus subtilis) fermented soy protein (FSP-B), respectively. Experimental diets for feeding trial were formulated to contain each soy protein sources at 8% level to corn-whey powder basal diet. There were 14 pigs per pen and 4 pens per treatment. Experimental diets were fed from 0 to 14 d after weaning and then a common commercial diet was fed from 15 to 35 d. Also for ileal digestibility studies, 18 pigs were assigned to 6 dietary treatments as N-free, SBM, SPC, HP300, FSP-A and FSP-B with T-canulation at distal ileum for 6 days. At $14^{th}$ d of experimental feeding, the ADG was significantly higher (p<0.05) in SPC fed diet as compared with others. Similar trend was noticed during the 15-35 d and overall study (0-35 d). All the processed soy protein sources tested in this experiment improved (p<0.05) growth than SBM during overall study. The nutrient digestibility of GE, DM, CP and Ca showed lower (p<0.05) values in SBM and FSP-A fed groups than SPC and FSP-B treatments. The apparent ileal digestibility of TEAA, non-TEAA and TAA showed lower (p<0.05) in SBM treatments compared with other soy protein sources. The true ileal digestibility of TEAA, non-TEAA and TAA were lower (p<0.05) in SBM fed group than SPC and HP300 treatments, and lower than FSP treatments though they didn't achieve significant difference (p>0.05). Villous height and crypt depth was not affected by dietary treatments. In conclusion, the growth and digestibility of nutrients in weaned pigs fed SPC was superior to others. Also FSP-A and FSP-B showed improved performance than those fed SBM.

Effects of Fermented Soy Protein on Nitrogen Balance and Apparent Fecal and Ileal Digestibility in Weaned Pigs

  • Yoo, J.S.;Jang, H.D.;Cho, J.H.;Lee, J.H.;Kim, I.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.22 no.8
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    • pp.1167-1173
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    • 2009
  • This study was conducted to evaluate the effects of providing fermented soy protein to weaned pigs on nitrogen balance and apparent fecal and apparent ileal digestibility (AID) of AA. Four weaned ((Yorkshire${\times}$Landrace)${\times}$Duroc) barrows (BW = 6.58${\pm}$0.98 kg), surgically fitted with a simple T-cannula approximately 15 cm prior to the ileo-cecal junction, were fed four diets according to 4${\times}$4 Latin square design. Diets were a basal diet supplemented with one of the following: 3% SDPP (spray dried plasma protein), 5% RBP (soy protein fermented by Lactobacillus spp.), 5% PSP (soy protein fermented by Aspergillus oryzae and Bacillus subtilis), and 2.5% RPP (2.5% RBP+2.5% PSP). No differences were observed in DM and N intakes among treatments. However, the level of urine excretion was greater in the RPP group than in the PSP group. Additionally, fecal DM excretion, fecal N concentration and fecal N excretion were increased in the RBP, PSP and RPP groups when compared with the SDPP group (p<0.05). Furthermore, total excretion was increased in the RPP group when compared with the PSP group (p<0.05). In addition, N absorption and the N absorption ratio were higher in the SDPP group than in the RPP group (p<0.05). Moreover, the DM and N digestibilities were lower in the RBP, PSP and RPP groups than in the SDPP group (p<0.05), and the ash and energy digestibilities were higher in the SDPP and RBP groups than in the PSP and RPP groups (p<0.05). However, no significant differences were observed in the DM, N, Ash, Ca, P or ileal digestibilities among treatments, although the energy digestibility was higher in the SDPP group than the RBP group (p<0.05). In addition, the apparent ileal digestibilities of essential amino acids (Arg, His, Iso, Leu, Lys, Phe, Thr, and Val) were significantly higher in the SDPP group than in the other groups (p<0.05), and the levels of Ala, Cys, Glu and Try were greater in the SDPP treatment group than the RBP, PSP and RPP groups (p<0.05). Additionally, the levels of Asp, Gly and Ser were higher in the SDPP group than the PSP and RPP groups, and the level of Pro was higher in the SDPP group than the RPP group (p<0.05). Finally, total non-essential amino acid and total amino acid digestibility were higher in the SDPP group than in the other treatments (p<0.05). Taken together, the results of this study indicate that animal protein is more bioavailable than plant protein. However, the N absorption ratio and ileal digestibility were found to be similar in the SDPP and RBP groups.

Characterization of a Multimodular Endo-β-1,4-Glucanase (Cel9K) from Paenibacillus sp. X4 with a Potential Additive for Saccharification

  • Lee, Jae Pil;Kim, Yoon A;Kim, Sung Kyum;Kim, Hoon
    • Journal of Microbiology and Biotechnology
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    • v.28 no.4
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    • pp.588-596
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    • 2018
  • An endo-${\beta}$-1,4-glucanase gene, cel9K, was cloned using the shot-gun method from Paenibacillus sp. X4, which was isolated from alpine soil. The gene was 2,994 bp in length, encoding a protein of 997 amino acid residues with a predicted signal peptide composed of 32 amino acid residues. Cel9K was a multimodular enzyme, and the molecular mass and theoretical pI of the mature Cel9K were 103.5 kDa and 4.81, respectively. Cel9K contains the GGxxDAGD, PHHR, GAxxGG, YxDDI, and EVxxDYN motifs found in most glycoside hydrolase family 9 (GH9) members. The protein sequence showed the highest similarity (88%) with the cellulase of Bacillus sp. BP23 in comparison with the enzymes with reported properties. The enzyme was purified by chromatography using HiTrap Q, CHT-II, and HiTrap Butyl HP. Using SDS-PAGE/activity staining, the molecular mass of Cel9K was estimated to be 93 kDa, which is a truncated form produced by the proteolytic cleavage of its C-terminus. Cel9K was optimally active at pH 5.5 and $50^{\circ}C$ and showed a half-life of 59.2 min at $50^{\circ}C$. The CMCase activity was increased to more than 150% in the presence of 2 mM $Na^+$, $K^+$, and $Ba^{2+}$, but decreased significantly to less than 50% by $Mn^{2+}$ and $Co^{2+}$. The addition of Cel9K to a commercial enzyme set (Celluclast 1.5L + Novozym 188) increased the saccharification of the pretreated reed and rice straw powders by 30.4% and 15.9%, respectively. The results suggest that Cel9K can be used to enhance the enzymatic conversion of lignocellulosic biomass to reducing sugars as an additive.

Expression and Characterization of Calcium- and Zinc-Tolerant Xylose Isomerase from Anoxybacillus kamchatkensis G10

  • Park, Yeong-Jun;Jung, Byung Kwon;Hong, Sung-Jun;Park, Gun-Seok;Ibal, Jerald Conrad;Pham, Huy Quang;Shin, Jae-Ho
    • Journal of Microbiology and Biotechnology
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    • v.28 no.4
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    • pp.606-612
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    • 2018
  • The enzyme xylose isomerase (E.C. 5.3.1.5, XI) is responsible for the conversion of an aldose to ketose, especially xylose to xylulose. Owing to the ability of XI to isomerize glucose to fructose, this enzyme is used in the food industry to prepare high-fructose corn syrup. Therefore, we studied the characteristics of XI from Anoxybacillus kamchatkensis G10, a thermophilic bacterium. First, the gene coding for XI (xylA) was inserted into the pET-21a(+) expression vector and the construct was transformed into the Escherichia coli competent cell BL21 (DE3). The expression of recombinant XI was induced in the absence of isopropyl-thio-${\beta}$-galactopyranoside and purified using Ni-NTA affinity chromatography. The optimum temperature of recombinant XI was $80^{\circ}C$ and measurement of the heat stability indicated that 55% of residual activity was maintained after 2 h incubation at $60^{\circ}C$. The optimum pH was found to be 7.5 in sodium phosphate buffer. Magnesium, manganese, and cobalt ions were found to increase the enzyme activity; manganese was the most effective. Additionally, recombinant XI was resistant to the presence of $Ca^{2+}$ and $Zn^{2+}$ ions. The kinetic properties, $K_m$ and $V_{max}$, were calculated as 81.44 mM and $2.237{\mu}mol/min/mg$, respectively. Through redundancy analysis, XI of A. kamchatkensis G10 was classified into a family containing type II XIs produced by the genera Geobacillus, Bacillus, and Thermotoga. These results suggested that the thermostable nature of XI of A. kamchatkensis G10 may be advantageous in industrial applications and food processing.

Synergistic Action Modes of Arabinan Degradation by Exo- and Endo-Arabinosyl Hydrolases

  • Park, Jung-Mi;Jang, Myoung-Uoon;Oh, Gyo Won;Lee, Eun-Hee;Kang, Jung-Hyun;Song, Yeong-Bok;Han, Nam Soo;Kim, Tae-Jip
    • Journal of Microbiology and Biotechnology
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    • v.25 no.2
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    • pp.227-233
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    • 2015
  • Two recombinant arabinosyl hydrolases, α-L-arabinofuranosidase from Geobacillus sp. KCTC 3012 (GAFase) and endo-(1,5)-α-L-arabinanase from Bacillus licheniformis DSM13 (BlABNase), were overexpressed in Escherichia coli, and their synergistic modes of action against sugar beet (branched) arabinan were investigated. Whereas GAFase hydrolyzed 35.9% of L-arabinose residues from sugar beet (branched) arabinan, endo-action of BlABNase released only 0.5% of L-arabinose owing to its extremely low accessibility towards branched arabinan. Interestingly, the simultaneous treatment of GAFase and BlABNase could liberate approximately 91.2% of L-arabinose from arabinan, which was significantly higher than any single exo-enzyme treatment (35.9%) or even stepwise exo- after endo-enzyme treatment (75.5%). Based on their unique modes of action, both exo- and endo-arabinosyl hydrolases can work in concert to catalyze the hydrolysis of arabinan to L-arabinose. At the early stage in arabinan degradation, exo-acting GAFase could remove the terminal arabinose branches to generate debranched arabinan, which could be successively hydrolyzed into arabinooligosaccharides via the endo-action of BlABNase. At the final stage, the simultaneous actions of exo- and endo-hydrolases could synergistically accelerate the L-arabinose production with high conversion yield.

Isolation and Identification of Three Pseudomonas koreensis Strains with Anti-microbial Activities Producing Inducers of the Expression of Egr-1 Gene (Egr-1 유전자의 발현 유도물질을 생산하는 항균성 저 영양 세균의 분리 및 동정)

  • Yoon, Sang-Hong;Kim, Dong-Gwan;Lee, Young-Han;Shin, Soon-Young;Kwon, Soon-Woo;Lee, Chang-Muk;Kang, Han-Chul;Koo, Bon-Sung
    • Microbiology and Biotechnology Letters
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    • v.39 no.2
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    • pp.119-125
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    • 2011
  • The Egr-1 gene is known to be a transcription factor for activating the expression of many tumor-repressing genes. In this study, three strains activating the promoter of the Egr-1 gene were selected, through the use of Egr-1 luciferase reporter assay and western blotting, from amongst approximately 3,800 oligotrophic bacteria isolated from the cultivated soils of various regions within Korea. These strains were identified as Pseudomonas koreensis on the basis of phylogenetic tree analysis of their 16S ribosomal DNA sequences and biochemical characteristics analyses using a variety of commercial kits (API 20NE, ID 32GN, API ZYM kits). In addition, we discovered that these strains produced anti-bacterial activity against Bacillus subtilis, Staphylococcus aureus and Listeria monocytogenes.

Problems and Verification System of Probiotics as Livestock-environment Improving Agent Produced and Circulated (축산 환경개선제로 생산.유통되는 생균제의 문제점 및 검증방안)

  • Lee, Eun-Young
    • Microbiology and Biotechnology Letters
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    • v.36 no.2
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    • pp.87-95
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    • 2008
  • Probiotics are live organisms that when administered in adequate amounts confer a health benefit on hosts. The administration of direct-fed microbials (DFM) such as lactobacilli and bacillus, may be a more direct approach to beneficially alter gastrointestinal microflora than altering dietary ingredients or supplementing with growth-promoting levels of antibiotics. It is apparent that microbes have an important influence on immune development and resistance to infection; that microbes are not static colonizers of our bodies, but are dynamic, symbiotic coresidents. And it can improve the surrounding environments; decrease the malodor caused by degrade the excrement. Recently, new paradigm such as environment protection and safe food have been settled. In domestic farm house, there is a great demand for probiotics as a substitute of antibiotics for the improvement of environmental quality and the production of a competitive goods. Probiotics circulated in a country have three categories: an animal medicine permitted by national veterinary research quarantine service (NVRQS), a support feed registered in city or country house, and not-registered goods. However, lots of unqualified goods were produced and circulated. And thus, it is in urgent need of evaluating the present situation and effect of probiotics. This study was conducted to evaluate the system of a probiotics as a livestock-environment improving agents for the alternation of antibiotics and quality control of it.

Change of Microflora in Livestock Manure during Composting Process (축산폐기물의 퇴비화 과정중 미생물상의 변동)

  • Whang, Kyun-Sook;Chang, Ki-Woon
    • Korean Journal of Soil Science and Fertilizer
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    • v.29 no.3
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    • pp.303-311
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    • 1996
  • The microflora changes of 10 water-controled treatments combined with livestock manures(pig, chicken) and bulking agents(sawdust, paper sludge) were investigated. The B/F values of the P-1 and C-1(65%, $H_2O$) treatments were 3571 and 5400 respectively, but those of the P-4 and C-4(50%, $H_2O$) treatments showed very low values, 667 and 334, respectively. The B/F values tended to increase with higher water content of the treatments. In the composting processes, the successions of microflora, adapting the compost environments, took place via fluctuating temperature. In the high temperature period, the numbers of mesophilic bacteria and fungi decreased, but that of the spore forming bacteria increased. However, the number of mesophilic bacteria inereased during the cold period. The B/F values of compost ranged 25-300, which indicates a decrease in the quantity of bacteria. The time required for the temperature of compost to reach $60^{\circ}C$ showed different patterns. There was no pathogenic microorganism in the treatments which reached a high temperature in a short period of time, but, in the treatments which reached a high temperature over a Long period of time, the pathgenic microorganism was not still alive.

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Impacts of Chemical Properties on Microbial Population from Upland Soils in Gyeongnam Province (경남지역 밭 토양 화학성분이 미생물 생태에 미치는 영향)

  • Lee, Young-Han;Ha, Sang-Keun
    • Korean Journal of Soil Science and Fertilizer
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    • v.44 no.2
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    • pp.242-247
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    • 2011
  • Soil management for environment-friendly agriculture depends on the effects of soil microbial activities and soil fertility. To improve soil health for the upland crops, this study evaluated a relationship between soil chemical properties and soil microbial diversities at 25 sites in upland soils in Gyeongnam Province. The average nutrients in the upland soils were 1.7 times for available phosphorous, 1.4 times for exchangeable potassium and 1.5 times for exchangeable calcium higher compared to recommend concentrations in the upland soils. We found a significant positive correlation between the soil organic matter and the soil microbial biomass C (p<0.01). Contents of organic matter and dehydrogenase in the inclined piedmont soils were significantly higher than those in the other topographical soils (p<0.05). In addition, concentrations of organic matter and microbial biomass C in the loam soils were significantly higher than in the silt loam soils (p<0.05). In principal component analyses of chemical properties and microbial populations in the upland soils, our findings suggested that available phosphorous should be considered as potential factor responsible for the clear upland soils differentiation. The soil organic matter was positive correlation with Bacillus sp. and fungi, whereas soil pH was also positive correlation with Pseudomonas sp. in upland soils.