• Journal of The Korean Society of Grassland and Forage Science
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• v.32 no.1
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• pp.59-74
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• 2012

Buffer solubility and protein fractionation were evaluated from the hays (timothy, alfalfa and klein) and straws (tall fescue and rice), and $In$ $vitro$ trial was conducted to examine the effect of buffer extraction on fermentation characteristics, degradability and gas ($CO_2$ and $CH_4$) production. Buffer soluble protein (SP) content and A fraction in total protein were highest in alfalfa hay as 61% and 41.77%, respectively while lowest in rice straw (42.8% and 19.78%, respectively). No difference was observed in B1 fraction among forages but B2 fraction was slightly increased in klein hay (12.34%) and tall fescue straw (10.05%) compared with other forages (6.34~8.85%). B3 fraction of tall fescue was highest as 38.49% without difference among other forages while C fraction was highest in rice straw. pH in incubation solution was higher in all forages after extraction than before extraction at 3h (P<0.01) and 6h (P<0.05), and pH from hays of timothy and alfalfa was higher than the other forages at 6h (P<0.05) and 12h (P<0.001). Regardless of extraction, ammonia-N concentration from alfalfa hay was increased at all incubation times and extraction effect was appeared only at 3h incubation time (P<0.01). Total VFA concentration from alfalfa hay was highest up to 24h incubation while those from tall fescue straw and rice straw were lowest. Buffer extraction decreased (P<0.01~P<0.001) the total VFA concentration. Acetic acid proportion was increased (P<0.001) before extraction of forages but no difference was found between forages. Propionic acid($C_3$) proportion was also increased(P<0.001) before extraction in all forages than in straws at 3h, 24h and 48h incubations, and $C_3$ from hays were mostly higher (P<0.05) than from straws. Butyric acid proportion, however, was not affected by extraction at most incubation times. Parameter 'a' regarding to the dry matter (DM) degradation was increase (P<0.001) in all forages before extraction, and was decreased (P<0.05) in tall fescue straw and rice straw compared with hays. Parameter 'b' was also increased (P<0.001) before extraction but no difference was found between forages. Effective degradability of DM (EDDM) was higher (P<0.001) before extraction in most forages except for rice straw. Buffer extraction decreased (P<0.05) all parameters (a, b, and c) regrading to the crude protein (CP) degradation but no difference was found between forages. Effective degradation of CP (EDCP) was lower (P<0.05) in straws than in hays. Parameters 'a' and 'b' regarding to the NDF degradation (P<0.01) and effective degradability of NDF (EDNDF, P<0.001) were also higher in forages before extraction than after extraction but no difference was found between forages. Buffer extraction reduced (P<0.05~P<0.001) $CO_2$ production from all the forages uo to 24h incubation and its production was greater (P<0.05~P<0.01) from hays than straws. Methane ($CH_4$) production was also greater (P<0.01~P<0.001) in all forages at all incubation times, and its production was greater (P<0.05) from hays than from straws at most incubation times. Based on the results of the current study, it can be concluded that buffer solubility and CP fractionation might be closely related with $In$ $vitro$ VFA concentration, degradability and gas ($CO_2$ and $CH_4$) production. Thus, measurement of buffer solubility and protein fractionation of forages might be useful to improve TMR availability in the ruminants.

• Microbiology and Biotechnology Letters
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• v.15 no.2
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• pp.129-134
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• 1987

Aspergillus sp. (MK-24) producing a biological active substance that inhibited the venom proteinase activity was isolated from soil. The substance also inhibited the activity of trypsin and coagulation of blood, but did not inhibit papain, $\alpha$-chymotrypsin and pepsin. The substance was partially purified from culture filtrate by precipitaion with acetone, and by chromatography of DEAE-Sepadex A-50 column and Amberlite IRC-50 ion exchange. The inhibitory substance was stable in the wide pH range from 2.0 to 12.0 at 37$^{\circ}C$, but not stable at $65^{\circ}C$ in the alkaline pH. Only 12% of the activity was decreased by the heat treatment at 10$0^{\circ}C$ for two hours. The inhibition on venom proteinase (Agkistrodon bromohoffi brevicaudus) was a mixed type. The inhibitory activity depended on the preincubation time and completely depressed by cupric, zinc and cobalt ions. The inhibition on the venom proteinase was appeared strongly on casein but not on ovalbumin or hemoglobin as a substrate.

• Proceedings of the KSAR Conference
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• 2003.06a
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• pp.70-70
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• 2003

조류의 배자는 포유류의 배자처럼 모체로부터 영양분을 공급받는 것이 아니라 알속에 저장되어 있는 영양물질로 발육한다. 배자의 발육은 대부분 체외에서 진행된다. 난자는 배란된 후 수정되어 난관팽대부에서 1세포기 수정란이 된다. 그 후 난관협부로 이동하여 최초로 분열이 일어나 배자의 발육이 진행되고, 산란시에는 세포수가 약6만여 개에 달한다. 따라서 수정란에 유전조작기법을 사용하기 위해서는 모체의 난관속에서 일어나는 배발생과 난각속에서 일어나는 개체발생을 위한 체외배양법과 대리난각 배양법이 확립되어 있어야 한다. 그와 같은 기술은 닭 수정란의 배 발생 관찰 및 분석과 유용한 물질을 생산하기 위한 형질전환 가금 연구에 중요한 기술로 사용되고 있다. 따라서 본 연구는 1세포기 닭 수정란의 체외배양과 대리난각 배양에 있어서 수정란의 조건과 대리난각의 조건이 배자의 생존율과 부화율에 미치는 영향을 조사하여 체외배양과 대리난각 배양에 의한 병아리 생산 효율을 제고하기 위한 기초자료를 얻고자 실시하였다. 주요 조사 항목은 수정란의 채란위치, 배자의 발생단계, 수정란의 무게, 대리난각용 계란의 보존 기간, 대리난각의 두께, 대리난각 두께의 감소율, 대리난각의 크기 등을 조사하여 배자의 생존율과 부화율에 미치는 영향에 대하여 조사하였다. 본 실험의 결과는 초기 발생이 빠른 배자가 생존율과 부화율이 높았으며, 본 실험에 사용한 대리난각용 계란의 보관기간이 짧을수록 배자의 생존율과 부화율이 높은 것으로 조사되었다(p<0.05). 그러나 대리난각용 계란의 크기와 대리난각의 두께 차이는 배자의 생존율과 부화율에 큰 영향이 없는 것으로 조사되었다.하였을 때 분할율은 68.0%였으며, 이중 12.0%가 상실배 또는 배반포로 발달하였다. 뿐만 아니라 10% FBS가 첨가된 TCM-199 배양액에 난관상피세포와 공배양을 실시하였을 경우는 72.0%가 분할하였으며, 이중 16.7%가 상실배 또는 배반포로 발달하였다. 이상의 결과로 볼 때 활성화 처리는 ionomycin과 6-DMAP 용액처리가 적합하며, 단위발생란의 체외배양은 보다 적합한 배양조건의 확립이 필요한 것으로 생각된다.icalcium lactate 공동배양은 체외배양에 별다른 영향을 미치지 못하는 것으로 나타났다.생산하는 것을 확인할 수 있었다.4시간에 등급별 회수율이 각각 GI(27.4%), GII(14.7%), GIII(43.2%), GIV(14.7%)로 나타났으며, 46~50시간에는 GI(12.0%), GII(12.0%), GIII(28.0%), GIV(48.0%)로 나타났다. 이상의 결과로 볼 때 미성숙 난자의 회수는 hCG 투여 후 29~34시간이 적합한 것으로 생각된다. 가금의 생산에 있어서 매우 효율적이고 주목할 만한 방법으로 사료된다. 것으로 나타났다.적외선.열풍 복합건조방법이 높게 나타나 이것은 곡물 표면에 원적외선 방사에의한 복사열이 전달되어 열장해를 받았기 때문으로 판단되며, 금후 더 연구하여 적정 열풍온도 및 방사체 크기를 구명해야 할 것이다.으로 보여진다 따라서 옻나무 유래 F는 포유동물의 생식기능에 중요하게 작용하는 것으로 사료된다.된다.정량 분석한 결과이다. 시편의 조성은 33.6 at% U, 66.4 at% O의 결과를 얻었다. 산화물 핵연료의 표면 관찰 및 정량 분석 시험시 시편 표면을 전도

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2002.11a
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• pp.77-77
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• 2002

본 연구는 체세포 및 체세포 핵이식란의 염색체 이상에 체세포의 계대의 영향성을 조사하고자 하였다. 한우 성축의 귀 조직으로 얻어진 세포를 공여세포를 체외성숙 후 제핵된 난자에 핵이식을 실시하였으며, 1.9kv/cm, 20$\mu\textrm{s}$/2times의 전기자극으로 융합후 5$\mu\textrm{g}$/ml의 ionomycin에서 4min, 1.9mM 6-DMAP에서 4h동안 배양함으로써 활성화를 유도하였다. 핵이식란은 CRlaa에서 4일간 배양 후 8 -cell단계에서 중기상의 유도를 위하여 상기 배양액 1ml당 0.05$\mu\textrm{g}$ colcemid에서 6-8시간 더 배양하였다. 이후, 6% Fetal bovine serum이 함유된 1% sodium citrate용액에 20분간 저장처리 후, methanol 5 : aceticacid 1 : distilled water 4로 1차, methanol 3: aceticacid 1 로 조성된 2차, methanol 4 : acetic acid 3 : distilled water 1의 3차고정액으로 1분간 재 침지시켰다. 고정 처리가 완료된 slide는 4% Giemsa용액으로 염색한 후 광학현미경 하에서 핵형 양상을 검경하였다. 체세포의 5계대에서는 684개의 spreads를 검경한 결과 염색체 수는 72%가 정상으로 60개이었고, 24%가 60개 이하였으며 4%가 60개 이상을 보였다. 10계대도 5계대와 비슷하여 71%가 정상, 26%가 60개 이하, 3%가 60개 이상이었고, 15계대에서는 55%가 정상이었고, 30%가 60개이하, 15%가 60개 이상을 보였다. 10계대 까지는 mixoploid의 비율의 변화가 없었으나 15계대에서 현저하게 늘어남을 볼 수 있었다. 또한 체외수정란과 핵이식란의 비교에서는 체외수정란은 250개의 spreads를 검경한 결과 염색체 수는 95.6%가 정상으로 60개이었고, 2.0%가 60개 이하, 2.4%가 60개 이상이었으나, 핵이식란은 204개를 검경하여 88%가 정상이었고, 4.9%가 60개이하, 7.1%가 60개 이상을 보임으로써, 핵이식란이 체외수정란에 비하여 염색체 이상의 비율이 높았다. 따라서 계대에 따라 체세포의 염색체이상의 비율이 상대적으로 증가하고, 체세포 핵이식에 따른 염색체 이상이 생길 수 있음을 알 수 있었다. (이 논문은 농림부 연구비에 의해 수행되었음)

• The Korean Journal of Food And Nutrition
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• v.23 no.1
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• pp.63-69
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• 2010

With the increase of the use of antibiotics and invasive procedures, infections caused by multidrug-resistant Acinetobacter baumannii(MRAB) are increasing. We screened the antibiotic producing strain B-51 for antibacterial activity against MRAB from the soils and studied the effects of culture medium on the antibiotic production of B-51. The medium conditions for maximum antibiotic productivity of B-51 was 2% glycerol, 0.5% soybean meal, 0.01% $CaCl_2$, 0.01% $MgSO_4{\cdot}7H_2O$ and 0.01% $KH_2PO_4$ at an initial pH of 6.0, at $30^{\circ}C$ for 76 h.

• Microbiology and Biotechnology Letters
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• v.20 no.3
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• pp.335-343
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• 1992

A Streptomyces strain No. 182-27, which produced amino acid antimetabolite, was isolated from soil. During the course of screening for new amino acid antimetabolites from the culture broths of Actinomycetes, we found that the strain produced a substance active against Gram-positive bacteria and its activity was reversed by L-Ieucine on the synthetic minimal agar medium in the culture broth. The morphological and cultural characteristics serve to identify the producing organism strain 182-27 as the Streptomyces, although the species of this strain should be resolved in further studies. Fermentation was carried out in the synthetic medium at $28^{\circ}C$ for 78 hours. The fermentation yield reached about 2 mg per liter of the broth. Purification was done by ion exchange resin, active carbon, silica gel column chromatography and obtained 20 mg of pure active substance from the 20 $\ell$ culture broth. The 182-27 substance was obtained as white powder, mp 18SoC. From the physicochemical characteristics of the substance, it was amino acid like substance but unknown about its chemical structure. It is active against some Gram-positive bacteria and reversed by L-Ieucine.

• KSBB Journal
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• v.6 no.2
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• pp.143-146
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• 1991

The liquifying $\alpha-amylase$ production from B. subtilis, A. oryzae and B. natto using wheat and rice bran as low cost culture medium was investigated. Among 3 strains, B. natto showed heights productivity of $\alpha-amylase$ in the outer wheat bran medium. And the optimum culture condition is pH 6.8 and $37^{\circ}C$ for the production of $\alpha-amylase$. The $\alpha-amylase$ activity of the crude enzyme and the purified enzyme are 256 unit/ml and 10,700 unit/ml, respectivitly. The $\alpha-amylase$ from B. natto cultrtured in outer wheat bran medium was purified into nearly a pure state(98.7%). And the molecular weight of the purified $\alpha-amylase$ was 34,000.

• KSBB Journal
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• v.19 no.1
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• pp.17-26
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• 2004

In this study the extracellular production of 5-aminolevulinic aicd (ALA) by recombinant E. coli BL2l (DE3) pLysS harboring the plasmid pFLS45 are investigated. Optimum concentrations of succinic acid and glycine for cell growth and ALA production were found to be 30 mM and 15 mM, respectively. Levulinic acid (LA) as an inhibitor of ALAD was added to the culture medium in the end of exponential cell growth phase and its optimum concentration was 30 mM. Growth of recombinant E. coli BL2l (DE3) pLysS (pFLS45) was largely dependent upon the pH value of culture medium. When the pH of culture medium was in the range of 6.0 and 6.5, high cell mass and ALA production were obtained. IPTG induction for the expression of the fusion gene did not enhance the production of ALA. Recombinant cell grew at 30't faster than at 37$^{\circ}C$, but ALA productivity was lower than at 37$^{\circ}C$. Repeated addition of glycine, succinic acid, and LA increased the production of ALA and the inhibition of intracellular ALA dehydratase activity, with up to 1.3 g/L ALA having been produced in the cultivation.

• Journal of Applied Biological Chemistry
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• v.61 no.4
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• pp.405-410
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• 2018

Pseudomonas tolaasii strain No. 6264 has been isolated from mushroom tissue and identified as one of the major pathogen causing brown blotch disease. It secretes peptide toxins, known as tolaasin and its analogue peptides. P. tolaasii 6264 has been used as a typical pathogenic strain to study the brown blotch disease for last 20 years after confirming its blotch-forming ability, hemolytic activity, and white line formation. In this study, the characteristics of P. tolaasii 6264 strain were analyzed and compared according to storage period. Strains of P. tolaasii 6264 stored annually since 2012 were cultured and their pathogenic characters were analyzed. When the 16S rRNA sequences were compared, all strains were divided into two groups. Pathogenic characters including hemolytic activity, blotch-forming ability, and white line test were also investigated. The strains, P. tolaasii 6264-15-2 and P. tolaasii 6264-17, had all three activities; however, the rest of stored strains showed only blotch-forming ability losing other pathogenic characters. Tolaasin peptides were purified from the bacterial cultures and analyzed by mass spectrometry. The strains, P. tolaasii 6264-15-2 and P. tolaasii 6264-17, secreted Tol I (1987 Da), Tol II (1943 Da), and its analogues (1973 Da, 2005 Da) while some of these peptides were not found in the media cultured other strains. These results indicate that the pathogenicity of P. tolaasii could be varied during the storage period.

• Journal of Animal Science and Technology
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• v.51 no.5
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• pp.369-378
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• 2009

Protein fractionation was evaluated from whole crop silages of rye (RS), wheat (WS), triticale (TS), oat (OS), barley (BS), and rice straw silage (RSS), and in vitro trial was carried out to examine the effect of silage and extraction of soluble protein on fermentation characteristics, total gas production and degradation. Soluble protein of silages was extracted with borate-phosphate buffer, and fermentation characteristics, gas production and degradation of silages were estimated by incubating anaerobically the mixed solution of strained rumen fluid and artificial saliva (1:1, v/v) containing dried and ground silages placed in nylon bag at $39^{\circ}C$ up to 48h. Soluble protein (SP) content was lowest for RSS as 2.11% in total CP compared to those for other silages. Highest A fraction (NPN) was observed from RS (74.33% of total CP) while those from TS and RSS were relatively low (48%). B2 fraction was relatively higher for RS, RSS and WS than for TS and BS. $B_3$ fraction was lowest in WS among silages. C fraction (27.07) in RSS was higher than in other silages (1.40~9.93%). pH in incubation solution was increased (P<0.01~P<0.001) for extracted silages up to 12h but decreased (P<0.01) at 48h for non-extracted ones. Contents of ammonia-N (P<0.001) and total VFA (P<0.01~P<0.001) were higher for non-extracted silages than for extracted ones. Acetate proportion was increased (P<0.001) in buffer extracted silages while those of propionate and butyrate were decreased (P<0.001) up to 24h incubation. Increased (P<0.001) total gas production was obtained from non-extracted silages up to 12h while gas production was increased (P<0.01) in extracted ones thereafter. In vitro degradation of dry matter and CP was increased (P<0.001) in non-extracted silages but that of neutral detergent fiber was increased (P<0.001) in extracted ones without difference among silages. Difference in mean values of degradability for each silage prior to- and post extraction with borate buffer, however, was not found among silages. It may be concluded that high NPN content of silages may reduce the protein availability in silages and borate buffer soluble components in silages can stimulate the early stage of fermentation.