• Title/Summary/Keyword: Protease production

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The Effects of Enzyme Complex on Performance, Intestinal Health and Nutrient Digestibility of Weaned Pigs

  • Yi, J.Q.;Piao, X.S.;Li, Z.C.;Zhang, H.Y.;Chen, Y.;Li, Q.Y.;Liu, J.D.;Zhang, Q.;Ru, Y.J.;Dong, B.
    • Asian-Australasian Journal of Animal Sciences
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    • v.26 no.8
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    • pp.1181-1188
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    • 2013
  • Two experiments were conducted to evaluate the effect of supplementing a corn-soybean meal-based diet with an enzyme complex containing amylase, protease and xylanase on the performance, intestinal health, apparent ileal digestibility of amino acids and nutrient digestibility of weaned pigs. In Exp. 1, 108 piglets weaned at 28 d of age were fed one of three diets containing 0 (control), 100, or 150 ppm enzyme complex for 4 wks, based on a two-phase feeding program namely 1 to 7 d (phase 1) and 8 to 28 d (phase 2). At the end of the experiment, six pigs from the control group and the group supplemented with 150 ppm enzyme complex were chosen to collect digesta samples from intestine to measure viscosity and pH in the stomach, ileum, and cecum, as well as volatile fatty acid concentrations and composition of the microflora in the cecum and colon. There were linear increases (p<0.01) in weight gain, gain: feed ratio and digestibility of gross energy with the increasing dose rate of enzyme supplementation during the whole experiment. Supplementation with enzyme complex increased the digesta viscosity in the stomach (p<0.05) and significantly increased (p<0.01) the concentrations of acetic, propionic and butyric acid in the cecum and colon. Enzyme supplementation also significantly increased the population of Lactobacilli (p<0.01) in the cecum and decreased the population of E. coli (p<0.05) in the colon. In Exp. 2, six crossbred barrows (initial body weight: $18.26{\pm}1.21$ kg), fitted with a simple T-cannula at the distal ileum, were assigned to three dietary treatments according to a replicated $3{\times}3$ Latin Square design. The experimental diets were the same as the diets used in phase 2 in Exp. 1. Apparent ileal digestibility of isoleucine (p<0.01), valine (p<0.05) and aspartic acid (p<0.05) linearly increased with the increasing dose rate of enzyme supplementation. In conclusion, supplementation of the diet with an enzyme complex containing amylase, protease and xylanase improved piglet performance. This is likely a result of improvement in nutrient digestibility, volatile fatty acid concentrations and bacteria ratio in the large intestine.

Characteristics and functional analysis of Bacillus strains from the fermented soybean products, Cheonggukjang (전통 발효 청국장으로부터 분리한 Bacillus 균주들의 특성 및 기능 분석)

  • Moon, Ji-Young;Kwon, Soon-Wo;Hong, Seung-Beom;Seok, Soon-Ja;Kim, Jeong-Seon;Kim, Soo-Jin
    • Korean Journal of Microbiology
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    • v.51 no.3
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    • pp.300-307
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    • 2015
  • For selecting Bacillus strains producing high-quality Cheonggukjang, 8 strains were isolated from the different Cheonggukjang samples. Seven of them exhibited the highest 16S rRNA gene sequence similarity value of over 99.9% to Bacillus subtilis subsp. subtilis and one of them showed the similarity to B. licheniformis. All the strains showed positive activities for amylase, cellulase, protease and lipase, and 6 strains are positive for fibrinolytic activity. To confirm the safety of the strains isolated from the samples of Cheonggukjang which are manufactured by traditional method, strains were analyzed for the presence of seven toxin genes of Bacillus cereus and results were found negative. And 7 strains did not produce at all or merely produce both histamine and tyramine, the representative biogenic amines. Biogenic amine degradation analysis by HPLC revealed that, most of them exhibited tyramine degradation activity. For Cheonggukjang fermented by artificial inoculation of selected strains, fermentation property, sensory test, volatile basic nitrogen production and metabolic profiles by $^1H-NMR$ were tested. Seven strains were confirmed to make high-quality Cheonggukjang.

Effects of Deodeok Contents on the Qualities of Quick Fermented Doenjang Type Product (더덕을 첨가하여 속성시킨 된장형 제품의 품질에 미치는 영향)

  • Hong, Seong-Cheol;Choi, Ki-Soon;Lee, Ho-Joon;Kwon, Dong-Jin
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.39 no.5
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    • pp.757-763
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    • 2010
  • The qualities of Deodeok Doenjang including physicochemical characteristics and sensory properties were investigated over the following range of Deodeok levels: 5, 10, 15 and 20% (all w/w). The strains used in the manufacturing of Doenjang were Bacillus sp. B-3 and Aspergillus sp. M-9 with the highest enzyme activities like amylase and protease. In case of Deodeok Doenjangs prepared with Bacillus sp. B-3, the amino-type nitrogen of Doenjang containing 10% (w/w) Deodeok was produced more than others during fermentation for 40 days. Amino-type nitrogen produced in Deodeok Doenjang prepared with Aspergillus sp. M-9 was more than one of Deodeok Doenjangs with Bacillus sp. B-3 and a commercial Doenjang. When Deodeok content exceeded 15% (w/w), higher content resulted in lesser amino-type nitrogen production. The results showed that Deodeok had influenced growth of Bacillus sp. and Aspergillus sp. Sensory evaluation showed that Deodeok Doenjangs containing 10% (w/w) Deodeok and 1% (w/w) Bacillus sp. B-3 and containing 20% (w/w) Deodeok and 1% (w/w) Aspergillus sp. M-9 were superior to the other Doenjangs tested.

Changes in Biological Qualities of Soy Grits Cheonggukjang by Fermentation with β-Glucosidase-Producing Bacillus Strains (β-Glucosidase 활성이 있는 균주 Bacillus Strains를 접종해 제조한 Soy Grits 청국장의 품질 특성)

  • Lee, Kyung Ha;Choi, Hye Sun;Hwang, Kyung A;Song, Jin
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.5
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    • pp.702-710
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    • 2016
  • The bioactivity of soy grits cheonggukjang was enhanced by fermentation using three ${\beta}$-glucosidase-producing Bacillus subtilis strains (HJ 18-9, HJ 25-8, and HJ 18-9+HJ 25-8) for 48 h at $37^{\circ}C$. The results indicate that protease, cellulase, and a-amylase activities significantly increased (P<0.05) with increasing fermentation time. In addition, the amino-type nitrogen content of B. subtilis-fermented soy grits cheonggukjang increased to 91.0~168.0 mg% after 48 h of fermentation. Among the isoflavones in soy grits cheonggukjang, contents of ${\beta}$-glucosides or acetyl-glucosides were reduced while aglycone content increased upon fermentation. In particular, soy grits cheonggukjang fermented with B. subtilis HJ18-9 and HJ25-8 showed the largest increases in aglycone content compared to complex treatment. These results provide useful information for development starter (single and complex) as well as for production of high quality fermented soybean food.

Physicochemical and Antioxidant Properties of Red Ginseng Marc Fermented by Bacillus subtilis HA with Mugwort Powder Addition (고초균 발효에 의한 홍삼박 발효물에 쑥 분말 첨가에 따른 물리화학적 및 항산화적 특성)

  • Jung, Hye-Won;Kim, Ji-Eun;Seo, Ji-Hyun;Lee, Sam-Pin
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.39 no.9
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    • pp.1391-1398
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    • 2010
  • Red ginseng marc (RGM) was fermented by the solid-state fermentation using Bacillus subtilis HA to produce biologically active compounds. The red ginseng marc fermented without mugwort possessed higher mucilage content (11.5%) and proteolytic activity (277.5 unit/g). The RGM fermented with 3% mugwort showed lower production of mucilage and protease activity whereas higher tyrosine content (581.3 mg%) and consistency index ($8.8\;Pa{\cdot}s^n$). The mucilage produced from fermented RGM contained $\gamma$-PGA with 1,100 kDa of molecular weight, and its yield was 15.9 g/kg. 70% ethanol extract from the RGM fermented with 3% mugwort had the highest DPPH radical scavenging effect ($IC_{50}$ value of 0.57 mg/mL), and the water extract showed the highest ABTS radical scavenging effect, indicating $IC_{50}$ value of 1.24 mg/mL. Overall, the RGM fermented by B. subtilis HA with mugwort contained various biologically active compounds having antioxidant effects.

Effects of Anti-Microbial Materials on Storages of Low Salted Doenjang (항균물질을 첨가한 저식염 된장의 저장성)

  • Kim, Jeong-Rye;Kim, Yon-Kyung;Kim, Dong-Han
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.11
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    • pp.1864-1871
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    • 2013
  • The effect of additives on the quality of low salted doenjang was investigated during storage. Amylase activity gradually decreased during storage and protease activity decreased after four weeks. The number of yeast was lower in the mustard or ethanol added groups without a difference in bacterial count. The L- and b-values decreased gradually during storage with lower total color difference (${\Delta}E$) in garlic added doenjang. Gas production was reduced in the ethanol or mustard added groups. Titratable acidity and acid values were low in the ethanol and ethanol-garlic added ones. A reducing sugar content was higher in the groups with added additives. Ethanol decreased to the largest extent in mustard added doenjang. Amino-type nitrogen decreased in ethanol added doenjang, whereas ammonia-type nitrogen was low in the ethanol or mustard added groups. The taste, flavor, and overall acceptability of doenjang were significantly higher in the ethanol or garlic added groups than in the other groups.

Selection of Biocontrol Agent of Tomato Gray Mold Disease from Flower and Pollinator Hive (토마토 꽃과 수정용 벌집으로부터 잿빛곰팡이병 방제 길항균주 선발)

  • Kim, Da-Ran;Lee, Jun-Taek;Kim, Hye sun;Jeon, Chang Wook;Kwak, Youn-Sig
    • The Korean Journal of Pesticide Science
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    • v.21 no.1
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    • pp.90-96
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    • 2017
  • Gray mold disease, cause by Botrytis cinerea, occurs severe damage on varieties of fruit and vegetable production, and have no a critical control method. In case of chemicals control, it is a trigger emergence of drug resistance strains due to using them continuously. In addition, the pathogen is difficult to control naturally because it is possible to survive regardless of host status. In this study, microorganisms were isolated from tomato flower and hive samples and in order to select suitable microbial control agents for tomato gray mold disease. During six-months study, we isolated 1,004 isolates from flower and 925 isolates from pollinator hive samples. Among them, 6 strains were selected based on result of antifungal activity test. The selected strains showed not only strong antifungal activity against gray mold pathogen, but also cellulase and protease enzyme activities. The selected strains were identified as Paenibacillus polymyxa. In plant assay, P. polymyxa prevented the gray mold disease occurrence near 75%.

Evaluation of Rhizobacterial Isolates for Their Antagonistic Effects against Various Phytopathogenic Fungi (식물 근권에서 분리한 미생물의 식물병원성 진균에 대한 길항효과 검정)

  • Kim, Yun Seok;Kim, Sang woo;Lamsal, Kabir;Lee, Youn Su
    • The Korean Journal of Mycology
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    • v.44 no.1
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    • pp.36-47
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    • 2016
  • This study was conducted to evaluate five different strains of rhizobacterial isolates viz. PA1, PA2, PA4, PA5 and PA12 for biological control against Colletotrichum acutatum, C. coccodes, C. gloeosporioides, C. dematium, Botrytis cinerea, Rhizoctonia solani, Sclerotinia minor and Fusarium sp. In vitro inhibition assay was performed on three different growth mediums, potato dextrose agar (PDA), tryptic soy agar (TSA), and PDA-TSA (1:1 v/v) for the selection of potential antagonistic isolates. According to the result, isolate PA2 showed the highest inhibitory effect with 65.5% against C. coccodes on PDA and with 96.5% against S. minor on TSA. However, the same isolate showed the highest inhibition with 58.5% against C. acutatum on PDA-TSA. In addition, an in vivo experiment was performed to evaluate these bacterial isolates for biological control against fungal pathogens. Plants treated with bacteria were analyzed with phytopathogens and plants inoculated with phytopathogens were treated with isolates to determine the biological control effect against fungi. According to the result, all five isolates tested showed inhibitory effects against phytopathogens at various levels. Mode of action of these rhizobacterial isolates was evaluated with siderophore production, protease assay, chitinase assay and phosphate solubilizing assay. Bacterial isolates were identified by 16S rDNA sequencing, which showed that isolates PA1 and PA2 belong to Bacillus subtilis, whereas, PA4, PA5, and PA12 were identified as Bacilus altitudinis, Paenibacillus polymyxa and Bacillus amyloliquefaciens, respectively. Results of the current study suggest that rhizobacterial isolates can be used for the plant growth promoting rhizobacteria (PGPR) effect as well as for biological control of various phytopathogens.

Changes in Quality Characteristics of Cheongkukjang added with Deodeok (더덕이 첨가된 청국장의 품질특성 변화)

  • Hong, Seong-Cheol;Kwan, Dong-Jin
    • Food Science and Preservation
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    • v.18 no.2
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    • pp.171-177
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    • 2011
  • In order to demonstrate the usefulness of Deodeok. the functions of Deodeok and the qualities of Cheongkukjang were investigated. DPPH (1.1-diphenyl-2-picryl-hydrazyl) radical scavenging activity of Deodeok extracted with 70% ethanol was higher than that of water extract. The direct antimutagenic effect of ethanol extract of Deodeok was examined by Ames test using Salmonella typimurium TA 98. The inhibition rates on ethanol extract at concentrations of 200, 1.000, 2.000, 3.000 and 4.000 ${\mu}g$/plate were 5.75, 31.38, 34.75, 53.50 and 83.75%, respectively. The inhibition rates of ethanol extract was higher over 2 times than that of water extract. The qualities including physicochemical and sensory properties of Deodeok Cheongkukjang were investigated over the following range of Deodeok levels; 5, 10, 15 and 20% (w/w). The strain used in Cheongkukjang manufacture was Bacillus sp. B-3 with the highest enzyme activities such as amylase and protease. During fermentation at $40^{\circ}C$ for 6 days. amino-type nitrogen content of Cheongkukjang containing 10% (w/w) Deodeok was more than others. When Deodeok content exceeded 15% (w/w), higher contents resulted in lesser amino-type nitrogen production. The results showed that Deodeok had influenced the growth of Bacillus sp. B-3. The L-value of Deodeok Cheongkukjang was decreased according to increasing the Deodeok contents. Sensory evaluation showed that Deodeok Cheongkukjang containing 10% (w/w) Deodeok was superior to other tested.

Enhanced Production of hCTLA4Ig through Increased Permeability in Transgenic Rice Cell Cultures (형질전환 벼 현탁세포 배양에서 투과성 증진을 통한 hCTLA4Ig의 생산성 증대)

  • Choi, Hong-Yeol;Cheon, Su-Hwan;Kwon, Jun-Young;Lim, Jung-Ae;Park, Hye-Rim;Kim, Dong-Il
    • KSBB Journal
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    • v.31 no.4
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    • pp.277-283
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    • 2016
  • In this system, rice cells were genetically modified to express human cytotoxic T-lymphocyte antigen 4-immunoglobulin (hCTLA4Ig) using RAmy3D promoter induced by sugar depletion. Even though the target protein fused with signal sequence peptide, plant cell wall can be a barrier against secretion of recombinant proteins. Therefore, hCTLA4Ig can be trapped inside cell wall or remained in intracellular space. In this study, to enhance the secretion of hCTLA4Ig from cytoplasm and cell walls into the medium, permeabilizing agents, such as dimethyl sulfoxide (DMSO), Triton X-100 and Tween 20, were applied in transgenic rice cell cultures. When 0.5% (v/v) of DMSO was added in sugar-free medium, intracellullar hCTLA4Ig was increased, on the other hand, the secreted extracellular hCTLA4Ig was lower than that of control. DMSO did not give permeable effects on transgenic rice cell cultures. And Triton X-100 was toxic to rice cells and also did not give enhancing permeability of cells. When 0.05% (v/v) Tween 20 was added in rice cell cultures, however, intracellular hCTLA4Ig was lower than that of control cultures. And the maximum 44.76 mg/L hCTLA4Ig was produced for 10 days after induction, which was 1.4-fold increase compared to that of control cultures. Especially, Tween 20 at 0.05% (v/v) showed the positive effect on the secretion of hCTLA4Ig though the decrease of intracellular hCTLA4Ig. Also, Tween 20 as a non-toxic surfactant did not affect the cell growth, cell viability and protease activity. In conclusion, secretion of hCTLA4Ig could be increased by enhancing permeability of cells regardless of the cell growth, cell viability and protease activity.