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Optimization of Mannitol Fermentation by Leuconostoc mesenteroides sp. strain JFY (Leuconostoc mesenteroides sp. strain JFY 균주에 의한 만니톨 발효 조건의 최적화)

  • Yoo Sun Kyun;Hur Sang Sun;Song Suckhwan;Kim Kyung Min;Whang Kyung Sook
    • Journal of Life Science
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    • v.15 no.3 s.70
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    • pp.374-381
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    • 2005
  • The production of functional foods providing health benefit is one of the fast growing fields in the food industry. Mannitol as GRAS (generally recognized as safe) is a functional food. Mannitol is about $70\%$ as sweet as sucrose and slowly and incompletely absorbed from the intestine, suppling only about one-half energy value of glucose. Commercially, the mannitol is synthesized by catalytic or electrochemical reduction of glucose. However, as strong demand for natural products increased, biological techniques have been developed for mannitol production. The object of this study was to determine the optimum conditions of mannitol fermentation by Leuconostoc mesenteroides sp. strain JFY isolated from fermented vegetables. The processes parameters such as pH, temperature, yeast extract concentration, and fructose concentration were optimized. The chosen ranges were 4.5 to 7.5 for pH, 22 to $34^{\circ}C$ for temperature, 0.05 to $2.0\%$ for yeast extract. and 5 to 350 g/L for fructose. The mineral medium used consisted of 3.0g $KH_2PO_4,\;0.01g\;FeSO_4{\cdot}H_2O,\;0.01g\;MnSO_4{\cdot}4H_2O,\;0.2g\; MgSO_4{\cdot}7H_2O,\;0.01g\;NaCl,\;and\;0.05g\;CaCl_2$ per 1 liter of deionized water. The optimum values of pH, temperature, yeast extract, and fructose concentration were obtained at about pH 6.5, temperature $28^{\circ}C$, yeast extract $0.5\%$ and fructose 30g/L. At optimum condition, the production of mannitol amounted to 31.6g/l. We hope that these findings are of particular importance for industrial application of mannitol production.

Mobility of Nitrate and Phosphate through Small Lysimeter with Three Physico-chemically Different Soils (소형 라이시메터시험을 통한 토양특성에 따른 질산과 인산의 이동성 비교)

  • Han, Kyung-Hwa;Ro, Hee-Myong;Cho, Hyun-Jun;Kim, Lee-Yul;Hwang, Seon-Woong;Cho, Hee-Rae;Song, Kwan-Cheol
    • Korean Journal of Soil Science and Fertilizer
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    • v.41 no.4
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    • pp.260-266
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    • 2008
  • Small lysimeter experiment under rain shelter plastic film house was conducted to investigate the effect of soil characteristics on the leaching and soil solution concentration of nitrate and phosphate. Three soils were obtained from different agricultural sites of Korea: Soil A (mesic family of Typic Dystrudepts), Soil B (mixed, mesic family of Typic Udifluvents), and Soil C (artificially disturbed soils under greenhouse). Organic-C contents were in the order of Soil C ($32.4g\;kg^{-1}$) > Soil B ($15.0g\;kg^{-1}$) > Soil A ($8.1g\;kg^{-1}$). Inorganic-N concentration also differed significantly among soils, decreasing in the order of Soil B > Soil C > Soil A. Degree of P saturation (DPS) of Soil C was 178%, about three and fifteen times of Soil B (38%) and Soil A (6%). Prior to treatment, soils in lysimeters (dia. 300 mm, soil length 450 mm) were tabilized by repeated drying and wetting procedures for two weeks. After urea at $150kg\;N\;ha^{-1}$ and $KH_2PO_4$ at $100kg\;P_2O_5\;ha^{-1}$ were applied on the surface of each soil, total volume of irrigation was 213 mm at seven occasions for 65 days. At 13, 25, 35, 37, and 65 days after treatment, soil solution was sampled using rhizosampler at 10, 20, and 30 cm depth and leachate was sampled by free drain out of lysimeter. The volume of leachate was the highest in Soil C, and followed by the order of Soils A and B, whereas the amount of leached nitrate had a reverse trend, i.e. Soil B > Soil A > Soil C. Soil A and B had a significant increase of the nitrate concentration of soil solution at depth of 10 cm after urea-N treatment, but Soil C did not. High nitrate mobility of Soil B, compared to other soils, is presumably due to relatively high clay content, which could induce high extraction of nitrate of soil matrix by anion exclusion effect and slow rate of water flow. Contrary to Soil B, high organic matter content of Soil C could be responsible for its low mobility of nitrate, inducing preferential flow by water-repellency and rapid immobilization of nitrate by a microbial community. Leached phosphate was detected in Soil C only, and continuously increased with increasing amount of leachate. The phosphate concentration of soil solution in Soil B was much lower than in Soil C, and Soil A was below detection limit ($0.01mg\;L^{-1}$), overall similar to the order of degree of P saturation of soils. Phosphate mobility, therefore, could be largely influenced by degree of P saturation of soils but connect with apparent leaching loss only more than any threshold of P accumulation.

Liquid culture condition of Tremella fuciformis mycelia (흰목이 균사 액체배양 조건)

  • Chang, Hyun-You;Lee, Chan;Choi, Sung-Woo;Yun, Jong Won
    • Journal of Mushroom
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    • v.6 no.1
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    • pp.27-31
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    • 2008
  • The optimization of submerged culture conditions for mycelial growth and exopolysaccharide (EPS) production in an edible mushroom Tremella fuciformis were studied in shake flasks and bioreactors. The temperature of $28^{\circ}C$ and pH 8 in the beginning of fermentation in agitated flasks was the most efficient condition to obtain maximum mycelial biomass and EPS. The optimal medium constituents were as follows (g l-1): glucose 20, tryptone 2, $KH_2PO_4$ 0.46, $K_2HPO_4$ 1 and $MgSO_4H_2O$ 0.5. The fungus was cultivated under various agitation and aeration conditions in a 5L stirred-tank bioreactor. The maximum cell mass and EPS production were obtained at a relatively high agitation speed of 200 rpm and at an aeration rate of 2 vvm. The flow behavior of the fermentation broth was Newtonian and the maximum apparent viscosity (35 cP) was observed at a highly aerated condition (2 vvm). The EPS productivity in an airlift reactor was higher than that in the stirred-tank reactor. The EPS was protein-bound polysaccharides consisted of mainly mannose, xylose, and fructose. The molecular weights of EPS were determined to be $1.3{\sim}1.5{\times}10^6$.

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Purification and Characterization of a Novel Alkaline Protease from Bacillus horikoshii

  • Joo, Han-Seung;Choi, Jang-Won
    • Journal of Microbiology and Biotechnology
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    • v.22 no.1
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    • pp.58-68
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    • 2012
  • An investigation was conducted on the enhancement of production and purification of an oxidant and SDS-stable alkaline protease (BHAP) secreted by an alkalophilic Bacillus horikoshii, which was screened from the body fluid of a unique Korean polychaeta (Periserrula leucophryna) living in the tidal mud flats of Kwangwha Island in the Korean West Sea. A prominent effect on BHAP production was obtained by adding 2% maltose, 1% sodium citrate, 0.8% NaCl, and 0.6% sodium carbonate to the culturing medium. The optimal medium for BHAP production contained (g/l) SBM, 15; casein, 10; $K_2HPO_4$, 2; $KH_2PO_4$, 2; maltose, 20; sodium citrate, 10; $MgSO_4$, 0.06; NaCl, 8; and $Na_2CO_3$, 6. A protease yield of approximately 56,000 U/ml was achieved using the optimized medium, which is an increase of approximately 5.5-fold compared with the previous optimization (10,050 U/ml). The BHAP was homogenously purified 34-fold with an overall recovery of 34% and a specific activity of 223,090 U/mg protein using adsorption with Diaion HPA75, hydrophobic interaction chromatography (HIC) on Phenyl-Sepharose, and ion-exchange chromatography on a DEAE- and CM-Sepharose column. The purified BHAP was determined a homogeneous by SDS-PAGE, with an apparent molecular mass of 28 kDa, and it showed extreme stability towards organic solvents, SDS, and oxidizing agents. The $K_m$ and $k_{cat}$ values were 78.7 ${\mu}M$ and $217.4s^{-1}$ for N-succinyl-Ala-Ala-Pro-Phe-pNA at $37^{\circ}C$ and pH 9, respectively. The inhibition profile exhibited by PMSF suggested that the protease from B. horikoshii belongs to the family of serine proteases. The BHAP, which showed high stability against SDS and $H_2O_2$, has significance for industrial application, such as additives in detergent and feed industries.

Studies on Microbial Utilization of Agricultural Wastes (Part 8) Pilot Plant Operation for the Production of Cellulosic Single Cell Protein (농산폐자원의 미생물학적 이용에 관한 연구 (제8보) 섬유질단세포단상질 생산의 시험공장조업)

  • Ko, Young-Hee;Lee, Kye-Joon;Bae, Moo
    • Microbiology and Biotechnology Letters
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    • v.5 no.3
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    • pp.119-125
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    • 1977
  • The cultivation of C. flavigena KIST 321, capable of utilizing cellulosic resources, was carried out in a 500 L fermentor by the batch process and the productivities of cellulosic SCP have been investigated by establishing the optimal conditions and levels of cellulosic material and others as medium components. The highest yield of the cell mass in the batch process was atttained under tile conditions at 30$^{\circ}C$, pH 7.4, 0.4∼0.6 VVM of aeration and at 130 rpm of agitation. According to the material balance of cellulosic SCP production using tile pretreated rice straw as a carbon source, more than 25 percent of rice straw on the base of drying weight was recovered in the form of cell mass.

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Characterization of Proteolytic Streptococcus sp. Isolated from Market Foods (시판식품에서 분리된 단백분해성이 강한 Streptococcus sp.의 특성)

  • CHANG Dong-Suck;LEE Jong S.
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.16 no.3
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    • pp.225-230
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    • 1983
  • The proteolytic bacteria were isolated from the market foods such as ground beef, cooked shrimp meat, perch fillet, oyster meat, beef with textured vegetable protein and fish digest distributed at supermarket in Corvallis, Oregon, U.S.A. Two hundred and twenty-eight strains($30.8\%$) have proteolytic activity from 740 strains isolated from the examined samples and the strongest proteolytic strain among them was identified as a Streptococcus sp. Its maximum growth was showed at about 6 hours culture at $37^{\circ}C$ with shaking incubator in the medium added $0.15\%$ potassium phosphate monobasic and $0.4\%$ potassium phosphate dibasic, while the strongest activity of its extracellular protease was observed after 7 hours culture. The exoenzyme produced by the Streptococcus sp. was observed as a metal chelator sensitive protease, which are strongly inhibited by EDTA and o-phenanthroline but not affected by phenylmethylsulfonylfluoride and p-hydroxymercuribenzoate.

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Studies on the Production of 5'-Nucleotides by Streptomyces spp. -Part 1. Isolation of 5'-phosphodiesterase Producing Microorganisms- (방사균(放射菌)에 의한 5'-Nucleotide류(類)의 생산(生産)에 관한 연구(硏究) -제 1 보(第 1 報) 5'-Phosphodiesterase생산균(生産菌)의 분리(分離)-)

  • Kim, Hong-Jip;Bae, Chong-Chan;Hwang, Kyu-Ln;Kong, Un-Young
    • Applied Biological Chemistry
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    • v.22 no.4
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    • pp.210-216
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    • 1979
  • RNA degrading bacteria were isolated from soil of Korea. One strain (no. JSC-114), having strong 5'-phosphodiesterase activity, was identified as belonging to the genus Streptomyces on the basis of taxonomic characteristics. The optimum conditions of 5'-phophosdiesterase production were found at $30^{\circ}C$ for 4 day in a medium containing 4.5% of soluble starch, 0.15% of peptone, 0.6% of yeast extract, 0.1% of $MgSO_4{\cdot}7H_2O$, 0.01% of $CaCl_2{\cdot}2H_2O$, 0.25% of $KNO_3$, and 0.5% of $KH_2PO_4$(pH 7.0). The maximum production rate of 5'-nucleotides from yeast RNA was 95% at $40-45^{\circ}C$ for 4hrs, and the products were identified as 5'-IMP, 5'-GMP, 5'-CMP and 5'-UMP(5.5 : 5.0 : 4.9 : 5.0).

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Effect of Cultural Conditions on Polysaccharide Production and its Physicochemical Properties in Cordyceps militaris

  • Kwon, Jeong-Seok;Hong, Eock-Kee
    • 한국생물공학회:학술대회논문집
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    • 2005.04a
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    • pp.140-143
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    • 2005
  • This study was concerned with the optimization of liquid culture conditions for mycelial growth and polysaccharide production and its physicochemical properties in Cordyceps militaris. The one factor at a time method was adopted to investigate the effects of medium composition, environmental factors and C/N ratio. Among the these varialbles, glucose 80g/L, yeast extract 10g/L, $MgSO_4{\cdot}7H_{2}O\;0.5g/L$, $KH_{2}PO_4\;0.5g/L$ were proved to be the most suitable carbon, nitrogen, and mineral sources, respectively. The optimal temperature, initial pH, working volume were identified to be $24^{\circ}C$, 7.0 and 100ml, respectively. Under the optimal conditions, the strategies in shake flask culture and 5L jar fermentor led to mycelial growth of 29.43 g/L, 28.88g/L and polysaccharide production of 2.53g/L, 6.38 g/L, respectively. Among the phisicochemical properties, relative concentrations(w/v) of total sugar, uronic acid, protein and hexoseamine were identified to be 74.07%, 1.13%, 0.91%, and 0.46%, respectively. The fraction of neutral and acidic polysaccharide were identified to be 81.9% and 18.1%, respectively.

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HPLC Chromatographic Methods for Simultaneous Determination of Pholcodine and Ephedrine HCI with Other Active Ingredients in Antitussive-Antihistamine Oral Liquid Formulations

  • Abdallah, Rokia M.
    • Natural Product Sciences
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    • v.12 no.1
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    • pp.55-61
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    • 2006
  • A description of simple, isocratic and precise reversed phase HPLC methods is given for simultaneous quantification of pholcodine and ephedrine hydrochloride together with either carbinoxamine maleate or terfenadine in antitussive-antihistaminic oral pharmaceutical formulations. Separations were carried out on X-Terra and symmetry shield C18 column $(250\;{\times}\;4.6\;mm,\;5\;{\mu}m)$. The used isocratic elution systems were either $0.02\;M\;KH_2PO_4-acetonitrile$ in the ratio of 75 : 25 and pH adjusted to 7.70 with orthophosphoric acid or sodium hydroxide, for syrup (method A), or 0.02 octanesulphonic acid sodium salt solution-acetonitrile-acetic acid in the ratio of 75 : 25 : 0.5 for suspension (method B). The elution of both mixtures was achieved with a flow rate of 1 ml/min. Detection was carried out by UV absorbance at wavelengths of 220 and 250 nm for syrup and suspension, respectively. The quantification of the components in synthetic mixtures and actual syrup and suspension were calculated using the internal standard technique with metoclopramide HCl and codeine phosphate as internal standards (IS), respectively. The methods, for both mixtures, were validated and met all the requirements for the quality control analysis recommended by FDA and ICH.

Some Cultural Characteristics of Bacillus megaterium YWO-5 Producing Bioflocculant for Wastewater Treatment (폐수처리용 생물응집제를 생산하는 Bacillus megaterium YWO-5의 배양특성)

  • Seo, Ho-Chan;Yeo, Sung-Jee;Cho, Hong-Yeon;Yang, Han-Chul
    • Microbiology and Biotechnology Letters
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    • v.27 no.1
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    • pp.80-85
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    • 1999
  • To develop bioflocculant for wastewater treatment, about 60 type culture strains and 450 strains isolated from natural sources were examined for screening their ability to flocculate the swine wastewater. Among them, YWO-5 showed the highest activity for NTU removal efficiency and was identified as Bacillus megaterium according to the cultural, morphological and physiological properties. The maximum production of the flocculant was achieved in culture medium containing 2% glucose, 0.05% soytone, 0.01% $CaCl_2$, 0.05% $KH_2PO_4$, and 0.05% yeast extract with initial pH 6.5 when cultured with rotary shaker controlled at 20$^{\circ}C$ and 150 rpm. With jar fermentor, the maximum production was reached to NTU removal efficiency of 93% after 3 days under the optimal conditions. The bioflocculant produced by Bacillus megaterium YWO-5 was effective on various suspended solids and organic wastewaters.

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