• Title/Summary/Keyword: Amino-acid

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Growth Performance and Meat Quality of Broiler Chickens Supplemented with Bacillus licheniformis in Drinking Water

  • Liu, Xiaolu;Yan, Hai;Lv, Le;Xu, Qianqian;Yin, Chunhua;Zhang, Keyi;Wang, Pei;Hu, Jiye
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.5
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    • pp.682-689
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    • 2012
  • A feeding trial was conducted to investigate effects of Bacillus licheniformis on growth performance and meat quality of broilers. Nine hundred one-d-old broiler chicks were randomly assigned to 3 experimental groups with three replicate pens of 100 broiler chicks. Three treatments were i) control, ii) basal diets supplemented with 1 ml of B. licheniformis for each in feed water per day iii) basal diets supplemented with 2 ml of B. licheniformis per chick in feed water per day. The supplementation of B. licheniformis significantly increased body weight in grower chickens (p<0.05), and significantly improved the feed conversion in 3 to 6 and 0 to 6 wk feeding period compared with the control group (p<0.05). Additionally, the supplement also resulted in increased protein and free amino acid contents, and decreased fat content in chicken breast fillet (p<0.05). Furthermore, improvement in sensory attributes was observed in broilers fed with the probiotic. In conclusion, B. licheniformis treatments resulted in a significant increase (p<0.05) in broiler productivity based on an index taking into account daily weight gain and feed conversion rate. Meanwhile, the probiotic contributed towards an improvement of the chemical, nutritional and sensorial characteristics of breast fillet. Overall, the study indicates that B. licheniformis can be used as a growth promoter and meat quality enhancer in broiler poultry.

Production and Separation of Anti-hypertensive Peptide during Chunggugjang Fermentation with Bacillus subtilis CH-1023 (청국장 발효과정 중 항고혈압성 peptide의 생산 및 분리)

  • Cha, Woen-Suep;Bok, Su-Kyung;Kim, Myoung-Uk;Chun, Sung-Sook;Choi, Ung-Kyu;Cho, Young-Je
    • Applied Biological Chemistry
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    • v.43 no.4
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    • pp.247-252
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    • 2000
  • As functionality investigation of Korean traditional soybean fermentation foods, an antihypertensive peptide was separated during Chunggugjang fermentation by Bacillus subtilis CH-1023 and investigated inhibitory effect against angiotensin converting enzyme. After incubation at $20^{\circ}C,\;30^{\circ}C,\;40^{\circ}C,\;50^{\circ}C,\;60^{\circ}C$ for the $0{\sim}72$ hrs, protein content, protease activity and angiotensin converting enzyme inhibitory rate were determined. The protein content and protease activity were increased and reached maximum at 60 hrs fermentation with $40^{\circ}C$ and decreased after the 60 hrs fermentation. The optimum condition for antihypertensive peptide from Chunggugjang was appeared for 60 hrs at $40^{\circ}C$. Crude extract of Chunggugjang was partially purified by Amicon YM-3 membrane filtration and Sephadex G-10, G-25 gel filtration. The purified peptide showed inhibitory rate of 94.3% with 0.5 mg peptide content. The most prominent amino acid composition of the peptide from Chunggugjang was alanine, followed by phenylalanine, histidine.

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Effect of Filtrate of Loess Suspension on Growth and Quality of Soybean Sprouts (콩나물의 생장과 품질에 미치는 황토 지장수의 효과)

  • Kang, Jeng-Yeol;Kang, Sun-Chul;Park, Shin
    • Applied Biological Chemistry
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    • v.43 no.4
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    • pp.266-270
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    • 2000
  • Using the filtrate of loess suspension, we cultivated soybean sprouts and investigated its effect on growth and quality in soybean sprouts. In comparison with soybean sprouts cultivated by tap water, the soybean sprouts cultivated by the filtrate of loess suspension at $20^{\circ}C$ showed increases in its weight by 11.4% and length by 14.9%. When cultivated at $25^{\circ}C$, the soybean sprouts by filtrate of loess suspension also showed increases in its weight by 9.9% and length by 11.0%. We compared inorganic element contents and pH level between the filtrate of loess suspension and tap water. Contents of Ca, Mg, K, Na, Zn, Mn, and Cu did not show any difference, while only P was higher in the tap water. pH value did not show much difference either. Consequently, it seemed that inorganic element contents and pH in the filtrate of loess suspension did not give any effect on the growth of the soybean sprouts. And also there was no any significant difference in inorganic element and amino acid contents in two kinds of soybean sprouts. However in a sensory test, the color and overall acceptability of the soybean sprouts cultivated by the filtrate of loess suspension showed better than the soybean sprouts cultivated by the tap water.

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Purification and Properties of Arylsulfatase of Serratia marcescens (Serratia marcens Arylsulfatase의 정제와 성질)

  • Yim, Moo-Hyun
    • Microbiology and Biotechnology Letters
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    • v.5 no.4
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    • pp.177-184
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    • 1977
  • Arylsulfatase catalyzes the release of SO$\sub$4//sup2/- from sulfate esters of simple phenols. Arylsolfatase occurs widely in animal tissues and in microorganisms including soil bacteria. Its widespread distribution suggests that it has a rather fundamental function and environmental meaning. It has been shown previously that arylsulfatase of Klebsiella was purified and characterized. A condition of arylsulfatase synthesis was tested with several strains of Serratia. Serratia marcescens could not utilize some sugars, such as xylose, rhamnose, glucosamine and arabinose hut glucose and mannitol as a sole carbon source. However, arylsulfatase synthesis was repressed by glucose but not by mannitol. The enzyme synthesis was repressed ob inorganic sulfate and methionine, and this repression was relieved by addition of tyramine. Arylsulfatase of S. marcescen was purified by fractionation with ammonium sulfate and followed by chromatographies on DEAE-Cellulose CM-Cellulose, and DEAE-Sephadex A-25. The molecular weight of arylsulfatase was determined to be 46,000 by SDS-Gel electrophoresis and 49,000 by Sephadex G-100 column chromatography. The enzyme showed some different properties with that of K. aerogenes. The activity was maximum at pH 6.8. The Km and Vmax values for p-nitrophenyl sulfate were 2.5${\times}$10$\^$-4/ M and 20 nmoles/min/mg protein, respectively. The enzyme showed high activities toward phenyl sulfate, ο-and p-nitro phenyl sulfates, and p-nitrocatechol sulfate. The inhibition of enzyme was strongly affected by hydroxylamine, inorganic fluoride, sulfide and phosphate, but by inorganic sulfate. Like Klebsiella arylsulfatase, tyramine, octopamine, and dopamine gave signifcant inhibitory effect.

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Secretory Expression and Characterization of an Acidic Endo-Polygalacturonase from Aspergillus niger SC323 in Saccharomyces cerevisiae

  • Zhou, Huoxiang;Li, Xi;Guo, Mingyue;Xu, Qingrui;Cao, Yu;Qiao, Dairong;Cao, Yi;Xu, Hui
    • Journal of Microbiology and Biotechnology
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    • v.25 no.7
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    • pp.999-1006
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    • 2015
  • The endo-polygalacturonase gene (endo-pgaA) was cloned from DNA of Aspergillus niger SC323 using the cDNA synthesized by overlapping PCR, and successfully expressed in Saccharomyces cerevisiae EBY100 through fusing the α-factor signal peptide of yeast. The fulllength cDNA consists of 1,113 bp and encodes a protein of 370 amino acids with a calculated molecular mass of 38.8 kDa. After induction by galactose for 48 h, the activity of recombinant endo-PgaA in the culture supernatant can reach up to 1,448.48 U/mg. The recombinant protein was purified to homogeneity by ammonium sulfate precipitation and gel filtration column chromatography and subsequently characterized. The optimal pH and temperature of the purified recombinant enzyme were 5.0 and 50℃, respectively. The Michaelis-Menten constant (Km) and maximal velocity (Vmax) of the enzyme for pectin were 88.54 μmol/ml and 175.44 μmol/mg/min, respectively. The enzyme activity was enhanced by Ca2+, Cu2+, and Na+, and strongly inhibited by Pb2+ and Mn2+. The pectin hydrolysates were mainly galacturonic acid and other oligo-galacturonates. Therefore, these characteristics suggest that the recombinant endo-PgaA may be of potential use in the food and feed industries.

Identification and Characterization of a New Alkaline SGNH Hydrolase from a Thermophilic Bacterium Bacillus sp. K91

  • Yu, Tingting;Ding, Junmei;Zheng, Qingxia;Han, Nanyu;Yu, Jialin;Yang, Yunjuan;Li, Junjun;Mu, Yuelin;Wu, Qian;Huang, Zunxi
    • Journal of Microbiology and Biotechnology
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    • v.26 no.4
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    • pp.730-738
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    • 2016
  • est19 is a gene from Bacillus sp. K91 that encodes a new esterase. A comparison of the amino acid sequence showed that Est19 has typical Ser-Gly-Asn-His (SGNH) family motifs and could be grouped into the SGNH hydrolase family. The Est19 protein was functionally cloned, and expressed and purified from Escherichia coli BL21(DE3). The enzyme activity was optimal at 60℃ and pH 9.0, and displayed esterase activity towards esters with short-chain acyl esters (C2-C6). A structural model of Est19 was constructed using phospholipase A1 from Streptomyces albidoflavus NA297 as a template. The structure showed an α/β-hydrolase fold and indicated the presence of the typical catalytic triad Ser49-Asp227-His230, which were further investigated by site-directed mutagenesis. To the best of our knowledge, Est19 is a new member of the SGNH hydrolase family identified from thermophiles, which may be applicable in the industrial production of semisynthetic β-lactam antibiotics after modification.

The Site-Directed A184S Mutation in the HTH Domain of the Global Regulator IrrE Enhances Deinococcus radiodurans R1 Tolerance to UV Radiation and MMC Shock

  • Zhang, Chen;Zhou, Zhengfu;Zhang, Wei;Chen, Zhen;Song, Yuan;Lu, Wei;Lin, Min;Chen, Ming
    • Journal of Microbiology and Biotechnology
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    • v.25 no.12
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    • pp.2125-2134
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    • 2015
  • IrrE is a highly conserved global regulator in the Deinococcus genus and contributes to survival from high doses of UV radiation, ionizing radiation, and desiccation. Drad-IrrE and Dgob-IrrE from Deinococcus radiodurans and Deinococcus gobiensis I-0 each share 66% sequence identity. However, Dgob-IrrE showed a stronger protection phenotype against UV radiation than Drad-IrrE in the D. radiodurans irrE-deletion mutant (ΔirrE), which may be due to amino acid residues differences around the DNA-binding HTH domain. Site-directed mutagenesis was used to generate a Drad-IrrE A184S single mutant, which has been characterized and compared with the ΔirrE mutant complemented strain with Drad-irrE, designated ΔirrE-E. The effects of the A184S mutation following UV radiation and mitomycin C (MMC) shock were determined. The A184S mutant displayed significantly increased resistance to UV radiation and MMC shock. The corresponding A184 site in Dgob-IrrE was inversely mutated, generating the S131A mutant, which exhibited a loss of resistance against UV radiation, MMC shock, and desiccation. qPCR analysis revealed that critical genes in the DNA repair system, such as recA, pprA, uvrA, and ddrB, were remarkably induced after UV radiation and MMC shock in the ΔirrE-IE and A184S mutants. These data suggested that A184S improves the ability against UV radiation and MMC shock, providing new insights into the modification of IrrE. We speculated that the serine residue may determine the efficiency of DNA binding, leading to the increased expression of IrrE-dependent genes important for protection against DNA damage.

Mutational Analysis of Thermus caldophilus GK24 ${\beta}$-Glycosidase: Role of His119 in Substrate Binding and Enzyme Activity

  • Oh, Eun-Joo;Lee, Yoon-Jin;Choi, Jeong-Jin;Seo, Moo-Seok;Lee, Mi-Sun;Kim, Gun-A;Kwon, Suk-Tae
    • Journal of Microbiology and Biotechnology
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    • v.18 no.2
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    • pp.287-294
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    • 2008
  • Three amino acid residues (His119, Glu164, and Glu338) in the active site of Thermus caldophilus GK24 ${\beta}$-glycosidase (Tca ${\beta}$-glycosidase), a family 1 glycosyl hydrolase, were mutated by site-directed mutagenesis. To verify the key catalytic residues, Glu164 and Glu338 were changed to Gly and Gln, respectively. The E164G mutation resulted in drastic reductions of both ${\beta}$-galactosidase and ${\beta}$-glucosidase activities, and the E338Q mutation caused complete loss of activity, confirming that the two residues are essential for the reaction process of glycosidic linkage hydrolysis. To investigate the role of His119 in substrate binding and enzyme activity, the residue was substituted with Gly. The H119G mutant showed 53-fold reduced activity on 5mM p-nitrophenyl ${\beta}$-D-galactopyranoside, when compared with the wild type; however, both the wild-type and mutant enzymes showed similar activity on 5mM p-nitrophenyl ${\beta}$-D-glucopyranoside at $75^{\circ}C$. Kinetic analysis with p-nitrophenyl ${\beta}$-D-galactopyranoside revealed that the $k_{cat}$ value of the H119G mutant was 76.3-fold lower than that of the wild type, but the $K_m$ of the mutant was 15.3-fold higher than that of the wild type owing to the much lower affinity of the mutant. Thus, the catalytic efficiency $(k_{cat}/K_m)$ of the mutant decreased to 0.08% to that of the wild type. The $k_{cat}$ value of the H119G mutant for p-nitrophenyl ${\beta}$-D-glucopyranoside was 5.l-fold higher than that of the wild type, but the catalytic efficiency of the mutant was 2.5% of that of the wild type. The H119G mutation gave rise to changes in optima pH (from 5.5-6.5 to 5.5) and temperature (from $90^{\circ}C\;to\;80-85^{\circ}C$). This difference of temperature optima originated in the decrease of H119G's thermostability. These results indicate that His119 is a crucial residue in ${\beta}$-galactosidase and ${\beta}$-glucosidase activities and also influences the enzyme's substrate binding affinity and thermostability.

Comparison of Microbial Diversity and Composition in the Jejunum and Colon of Alcohol-Dependent Rats

  • Fan, Yang;Ya-E, Zhao;Ji-dong, Wei;Yu-fan, Lu;Ying, Zhang;Ya-lun, Sun;Meng-Yu, Ma;Rui-ling, Zhang
    • Journal of Microbiology and Biotechnology
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    • v.28 no.11
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    • pp.1883-1895
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    • 2018
  • Alcohol dependence is a global public health problem, yet the mechanisms of alcohol dependence are incompletely understood. The traditional view has been that ethanol alters various neurotransmitters and their receptors in the brain and causes the addiction. However, an increasing amount of experimental evidence suggests that gut microbiota also influence brain functions via gut-to-brain interactions, and may therefore induce the development of alcohol use disorders. In this study, a rat model of alcohol dependence and withdrawal was employed, the gut microbiota composition was analyzed by high-throughput 16S rRNA gene sequencing, and the metagenome function was predicted by PICRUSt software. The results suggested that chronic alcohol consumption did not significantly alter the diversity and richness of gut microbiota in the jejunum and colon, but rather markedly changed the microbiota composition structure in the colon. The phyla Bacteroidetes and eight genera including Bacteroidales S24-7, Ruminococcaceae, Parabacteroides, Butyricimonas, et al were drastically increased, however the genus Lactobacillus and gauvreauii in the colon were significantly decreased in the alcohol dependence group compared with the withdrawal and control groups. The microbial functional prediction analysis revealed that the proportions of amino acid metabolism, polyketide sugar unit biosynthesis and peroxisome were significantly increased in the AD group. This study demonstrated that chronic alcohol consumption has a dramatic effect on the microbiota composition structure in the colon but few effects on the jejunum. Inducement of colonic microbiota dysbiosis due to alcohol abuse seems to be a factor of alcohol dependence, which suggests that modulating colonic microbiota composition might be a potentially new target for treating alcohol addiction.

Effects of Composted Liquid Manure and Microbial Agent Types on Growth and Thatch Decomposing of Creeping Bentgrass (가축분뇨발효액비와 미생물제제 종류별 시용에 따른 크리핑 벤트그래스의 생육과 토양중 대취분해에 미치는 영향)

  • Lim, Ji Yeon;Ham, Suon Kyu;Lee, Yeong Min;Cha, Young Gi
    • Journal of the Korea Organic Resources Recycling Association
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    • v.22 no.4
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    • pp.54-61
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    • 2014
  • This study was conducted to investigate the effect of Actinomyces sp. and Bacillus sp., United States granular microorganisms and Japan granular microorganisms on turfgrass growth and thatch decomposing of creeping bentgrass in golf course by measuring turf color index, chlorophyll index, thatch content of soil, root length, turf density and chemical properties and thatch content of soil. Fertilizer treatment was designed as follows; control(CF; compound fertilizer), microorganism medium(M; CF+M), microorganism medium and livestock manure fertilizer(M-L; CF+M+LMF), microorganism medium, livestock manure fertilizer and amino acid liquid fertilizer(M-L-A; MM+LMF+ALF), United States granular microorganisms(USGM; CF+USGM), Japan granular microorganisms(CF+JGM). Soil properties investigated after experiment was scarcely affected by applied fertilizers in root zone of creeping bentgrass. The turf color index and chlorophyll index of M, M-L, M-L-A, USGM, JGM treatment were higher than those of CF. The turfgrass root in M-L treatment was longer than others. The thatch content of soil in M treatment was longer than others. The thatch content of M was decreased than that of CF by 6.8%. These was suggested that application of M induced the development of quality and growth of creeping bentgrass by assisting turfgrass growth and thatch decomposing.