• Journal of Microbiology and Biotechnology
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• 제22권11호
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• pp.1518-1522
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• 2012

Leuconostoc mesenteroides is a heterofermentative Grampositive bacterium that plays key roles in fermentation of foods such as kimchi, sauerkraut, and milk, leading to the production of various organic acids and aromatic compounds. To study the microbiological and genomic characteristics of L. mesenteroides, we have developed a new chemically defined minimal medium by using the single omission technique. During the exponential cell growth, this species required glutamine, methionine, valine, and nicotinic acid as essential nutrients and 8 amino acids (arginine, cysteine, histidine, leucine, phenylalanine, proline, threonine, and tryptophan), 5 vitamins (ascorbic acid, folic acid, inosine, calcium panthothenate, and thiamine), and others (manganese, magnesium, adenine, uracil, and Tween 80) as supplemental nutrients. This medium is useful to study the metabolic characteristics of L. mesenteroides and to explain its role in food fermentation.

• 한국해양바이오학회지
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• 제10권1호
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• pp.18-25
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• 2018

Chlorella is quantitatively and qualitatively high in protein with balanced essential amino acid profiles, vitamins and minerals. ${\gamma}-Aminobutyric$ acid (GABA) is broadly distributed in nature and fulfills multi-physiological functions including effect such as a health-promoting functional compound. To improve the GABA production, Chlorella protothecoides were grown through the modified mixtrophic culture medium containing 2L of sterilized bristol medium with 0.01% urea and 4.0% glucose in a 5L fermenter. The results showed that nineteen kinds of amino acid including GABA at C. protothecoides sample were analyzed using high performance liquid chromatography (HPLC). Glutamic acid in total concentration (%) of amino acid is the most abundant amino acid (33.10%), followed by alanine (20.48%) and GABA (17.48%). Three amino acids including GABA were responsible for more than 70% total concentration in C. protothecoides sample including eight essential and nine non-essential amino acids: aspartic acid, asparagine, serine, glutamine, histidine, glycine, threonine, arginine, tyrosine, valine, methionine, tryptophan, phenylalanine, isoleucine, leucine, lysine. As a result of this experiment, it is expected that Chlorella will be developed to a critical product having high value as, GABA, functional food materials.

• Asian-Australasian Journal of Animal Sciences
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• 제15권12호
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• pp.1714-1718
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• 2002

Twelve prepubertal Karan Fries heifers (15 months, $167.7{\pm}13.5kg$) were divided into two equal groups. Group 1 was fed as per NRC requirements and group 2 was fed 20% more protein than group 1 heifers. The experimental feeding was continued until the onset of puberty in both the groups. Blood samples were collected at fortnightly intervals and analyzed for amino acids using HPLC. Group 1 and 2 heifers required $178.6{\pm}33.8$ and $152.8{\pm}33.2$ days of experimental feeding to exhibit first estrus resulting in total age at puberty as $639.4{\pm}27.3$ and $618.6{\pm}24.6$ days in the two groups respectively. The concentration of total amino acids averaged 4.40 and 4.89 mmol/l and those of non-essential amino acids (NEAA) was 2.32 and 2.49 mmol/l in groups 1 and 2, respectively. The concentration of plasma essential amino acids i.e. histidine, threonine, valine, methionine, isoleucine, leucine and phenylalanine were higher (p<0.01) in group 2 than group 1. Plasma concentration of large neutral amino acids (LNAA) was significantly higher in group 2 (1.28 mmol/l) than in group 1 (1.12 mmol/l). Increased levels of leucine, isoleucine and valine are implicated in increased follicular growth and development in prepubertal heifers and resulted in a 26 day earlier attainment of puberty by 26 days in an experimental period of six months in group 2 heifers. Increased concentrations of aspartate and tyrosine in group 2 heifers might be associated with the release of GnRH from the hypothalamus influencing LH release from anterior pituitary in such animals. It is therefore evident that increased availability of certain amino acids in heifers fed high protein diet might have led to early onset of puberty.

• Journal of Microbiology and Biotechnology
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• 제13권3호
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• pp.415-421
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• 2003

A bacterium, Pseudomonas fluorescens 2112, that is antagonistic against a red-pepper blight-causing fungus, Phytophthora capsici, was isolated from the local soil of Gyongju, Korea. This strain formed an orange-colored clear halo zone on chrome azurol S (CAS) blue agar, suggesting the production of a siderophore in addition to an antifungal antibiotic. The optimal culture conditions for siderophore production by P. fluorescens 2112 were 30-h cultivation at $25^{\circ}C$ and pH 6.5 in King's B medium. The presence of $20{\mu}g/ml\;of\;Fe^3+$ ion or EDDHA promoted the production of siderophore in King's B medium. The siderophore was purified from culture broth by CM-Sephadex C-25 and Sephadex G-25 column chromatographies. The UV spectra of the purified siderophore was the same as that of pyoverdins or pseudobactins. The molecular mass was 1,958 Da determined by FAB-rlass spectrometer, and the amino acid composition analysis showed that the purified siderophore consisted of glycine/threonine/serine/glutamic acid/alanine/lysine with the molar ratio of 3:2:1:1:1:1, DL-Threo-${\beta}$-hydroxyaspartic acid and $N^{\delta}$-hydroxyornithine, two of the essential constituents of pyoverdin, were also found. The purified siderophore pyoverdin showed strong in vitro and in vivo antagonistic activities against phytophthora blight-causing P. capsici. Especially in an in vivo pot test, the siderophore protected red-pepper Capsicum annum L. very well from the attack of P. capsici. These results indicated that the purified siderophore of P. fluorescens 2112 played a critical role in the biocontrol of the red-pepper blight disease, equivalent to treatment by P.fluorescens 2112 cells.

• Asian-Australasian Journal of Animal Sciences
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• 제13권6호
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• pp.817-823
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• 2000

Four groups of six growing gilts each were fed ad libitum diets composed of cereals and soyabean (SBM) or rapeseed (RSM) meal and containing two levels of crude protein: high - 18.0% (RSM-H) and 16.9% (SBM-H) or low - 15.6% (RSM-L) and 15.1% (SBM-L). The diets were balanced by supplementation with crystalline amino acids and contained apparent ileal digestible lysine, methionine, threonine and tryptophan in proportions (1.00:0.32:0.57:0.18) according to CVB (1995). Voluntary feed intake, weight gain and slaughter and chemical body composition of animals were assessed. Protein and energy balances from 25 to 70 kg body weight were calculated by the comparative slaughter method. Protein source had a significant effect on voluntary feed intake; it was 0.12 kg/d lower in pigs fed the SBM than RSM-diets. Pigs fed on the SBM-L diet consumed the least amount of feed (2.17 kg). Daily gain (average, $900{\pm}12.59g$) and feed conversion ratio ($2.54{\pm}0.04kg/kg$) were not statistically affected by source (SMB and RSM) and protein level (high and low). In empty body similar amounts (g/kg) of protein (163 g), water (635 g) and ash (28 g) were found. However, pigs fed the RSM-L diet were fatter than those fed the SBM-L diet (188 vs. 161 g/kg). No statistical differences were observed in daily protein deposition, which on average amounted to $142{\pm}11g$, or carcass characteristics. An improvement of crude protein utilization by 6.3 percentage units was found by decreasing the protein concentration in the diets. Heat production in the body was not significantly affected by the treatments.

• Genomics & Informatics
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• 제13권1호
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• pp.2-6
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• 2015

Pathogenic bacteria survive in iron-limited host environments by using several iron acquisition mechanisms. Acinetobacter baumannii, causing serious infections in compromised patients, produces an iron-chelating molecule, called acinetobactin, which is composed of equimolar quantities of 2,3-dihydroxybenzoic acid (DHBA), L-threonine, and N-hydroxyhistamine, to compete with host cells for iron. Genes that are involved in the production and transport of acinetobactin are clustered within the genome of A. baumannii. A recent study showed that entA, located outside of the acinetobactin gene cluster, plays important roles in the biosynthesis of the acinetobactin precursor DHBA and in bacterial pathogenesis. Therefore, understanding the genes that are associated with the biosynthesis and transport of acinetobactin in the bacterial genome is required. This review is intended to provide a general overview of the genes in the genome of A. baumannii that are required for acinetobactin biosynthesis and transport.

• Journal of Microbiology and Biotechnology
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• 제25권7호
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• pp.1015-1025
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• 2015

The development of diverse polymyxin derivatives is needed to solve the toxicity and resistance problems of polymyxins. However, no platform has generated polymyxin derivatives by genetically engineering a polymyxin synthetase, which is a nonribosomal peptide synthetase. In this study, we present a two-step approach for the construction of engineered polymyxin synthetases by substituting the adenylation (A) domains of polymyxin A synthetase, which is encoded by the pmxABCDE gene cluster of Paenibacillus polymyxa E681. First, the seventh L-threonine-specific A-domain region in pmxA was substituted with the L-leucine-specific A-domain region obtained from P. polymyxa ATCC21830 to make polymyxin E synthetase, and then the sixth D-leucine-specific A-domain region (A_6-D-Leu-domain) was substituted with the D-phenylalanine-specific A-domain region (A_6-D-Phe-domain) obtained from P. polymyxa F4 to make polymyxin B synthetase. This step was performed in Escherichia coli on a pmxA-containing fosmid, using the lambda Red recombination system and the sacB gene as a counter-selectable marker. Next, the modified pmxA gene was fused to pmxBCDE on the chromosome of Bacillus subtilis BSK4dA, and the resulting recombinant strains BSK4-PB and BSK4-PE were confirmed to produce polymyxins B and E, respectively. We also succeeded in constructing the B. subtilis BSK4-PP strain, which produces polymyxin P, by singly substituting the A_6-D-Leu-domain with the A_6-D-Phe-domain. This is the first report in which polymyxin derivatives were generated by genetically engineering polymyxin synthetases. The two recombinant B. subtilis strains will be useful for improving the commercial production of polymyxins B and E, and they will facilitate the generation of novel polymyxin derivatives.

• Asian-Australasian Journal of Animal Sciences
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• 제14권5호
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• pp.655-667
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• 2001

This experiment was conducted to investigate the effect of feeding a low CP diet supplemented with synthetic amino acids on performance, nutrient utilization and carcass characteristics of finishing pigs fed under a three-phase feeding regimen. Ninety-six finishing pigs (Landrace$\times$Large White$\times$Duroc), $55.75kg{\pm}0.65$ of initial body weight, were blocked by weight and sex and allotted to four dietary treatments in a randomized block design. There were six pens per treatment and four pigs per pen. Pigs were fed a 16%-14%-12% CP (for phase I-II-III, respectively), sequence of diets. Dietary treatments were 1) Control, 2) Con+L (a sequence of diets reduced in CP by l percentage unit with lysine (L) supplementation, 3) Con+LMT (a sequence of diets reduced in CP by 2 percentage unit with LYS, methionine (MET) and threonine (THE) supplementation) and 4) Con+LMTT (a sequence of diets reduced in CP by 3 percentage unit with LYS, MET, THR and tryptophan (TRP) supplementation). The finishing period (55 to 105 kg) was divided into three phases (55 to 72 kg, 72 to 90 kg and 90 to 105 kg). Pigs fed either the control or Con+L diet grew faster (p<0.05) than pigs fed the Con-LMT or Con+LMTT diet. There was no difference in ADFI among dietary treatments. Phosphorus (P) digestibility was lowest in the control group and highest in the Con+LMTT group (p<0.05). Within each phase, no significant differences in dry matter (DM) and CP digestibilities were found. Although some amino acid digestibilities were affected by dietary treatments, digestibilities of essential amino acids (EAA), non-essential amino acids (NEAA) and total amino acid were not significantly influenced by dietary treatments. For the entire experiment periods, Con+L, Con+LMT and Con+LMTT treatments resulted in 13.4, 18.8 and 21.6% lower total N excretion compared with the control. Con+LMT and Con+LMTT treatments showed significantly lower BUN concentration compared with the control and Con+L treatment (p<0.05), but there was no significant difference in BUN concentration between pigs fed the control and Con+L treatment or between pigs fed Con+LMT and Con+LMTT treatments (p>0.05). Carcass length, backfat thickness and carcass grade were not significantly affected by dietary treatments (p>0.05). In conclusion, reducing dietary CP level by 1 percentage unit and supplementing only LYS at each phase could be a very beneficial feeding strategy for finishing pigs fed under a three phase feeding regimen in terms of both environmental and economical aspects.

• 한국미생물·생명공학회지
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• 제37권2호
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• pp.140-146
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• 2009

Cyclosporin A 분자의 구성 아미노산 가운데 분지 아미노산인 valine, leucine, isoleucine, threonine 등에 대한 첨가 효과를 확인한 결과, L-valine 첨가구는 비첨가구보다 cyclosporin A 역가는 3.7배, cyclosporin A 구성비율은 약 40%로 가장 크게 증가하였다. 그리고 L-valine의 최적 농도는 0.75%로서 비첨가구에 비하여 cyclosporin A 역가는 3배, cyclosporin A 구성비율은 30% 증가하였다. 그러나 L-valine 1% 단독 첨가구보다 L-valine 0.5%, L-leucine 0.5% 혼합구가 cyclosporin A 역가에서는 약 16% 우수하였고, cyclosporin A 구성비율에서는 약 5% 증가하였다. 또한 다양한 유기질소원의 첨가 효과를 확인한 결과, meat peptone인 Bacto-peptone 첨가구는 유기질소원 비첨가구보다 cyclosporin A의 역가는 약 2배, cyclosporin A 구성비율은 약 13% 증가하였고, Bacto-peptone의 최적농도는 1%인 것으로 확인되었다. 그러나 Casiton, Tryptone, NZ-amine 등 우유 유래 유기질소원 첨가구의 경우, 비첨가구보다 cyclosporin A 역가가 오히려 약 10-20% 감소하는 현상을 보여 cyclosporin A 생합성을 억제하는 것으로 확인되었다. 본 연구에서 최적화된 배지 조건에서, 모균주인 T. inflatum ATCC 34921와 UV 조사로 얻어진 4종의 cyclosporin A 고생산 변이주 4종의 생산성을 비교한 결과, 변이주 YHC-004의 cyclosporin A 역가는 3,430 mg/L로 모균주인 ATCC 34921보다 약 7.1배로 증가하였지만, 모균주와 변이주 4종의 cyclosporin A 구성비율은 약 93-94% 범위에서 비슷한 값을 보였다.

• Asian-Australasian Journal of Animal Sciences
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• 제24권11호
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• pp.1623-1628
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• 2011

The effects of dietary lysine (Lys) at a fixed ratio to crude protein (CP) and essential amino acids (EAA) on early growth response of broilers were studied. Four diets were formulated to contain similar metabolizable energy (ME, 2,950 kcal/kg) but contained graded levels of incremental Lys (1.1, 1.2, 1.3 and 1.4%) while also increasing the dietary CP and EAA (methionine, methionine+ cystine, threonine and tryptophan) to maintain a constant ratio with Lys. Each diet was fed at random to 10 replicates of 6 chicks each throughout the experimental period (1-21 d). At the lowest concentration of Lys of 1.1% (19.04% CP), body weight gain (BWG) was lowest and feed conversion ratio (FCR) was poorest. The BWG increased and FCR decreased linearly as dietary Lys increased upto 1.3% (22.5% CP). Lowest feed consumption was observed in the dietary group that contained 1.1% Lys (19.04% CP) in the diet. Increasing the concentration of Lys to 1.2% (20.77% CP), significantly increased the feed consumption. The concentrations of protein, calcium, phosphorus and cholesterol in serum were not influenced by the variation in Lys contents in the diet. The humoral immune response as measured by antibody titre in response to SRBC inoculation was significantly lower in the diets containing 1.1% Lys compared to 1.4%. It is concluded that the Lys requirement of broilers is 1.3% (22.5% CP) during 0 to 21 days of age for eliciting optimum performance when a fixed ratio of Lys to CP (1:17.31) and essential AA is maintained (1:0.47 Met; 1:0.56 Thr; 1:0.17 Try).