Brewer's yeast extract can be used as a good substrate for the culture of Lentinula edodes Mycelia(LEM). We found that it was better to filter the extract through three kinds of sieves and then to heat for hydrolysis and concentration at $90^{\circ}C$ prior to use. Also the maximum condition for the growth of LEM was investigated. We found that addition of inorganic salts such as calcium enhanced the growth of LEM. On the other hand, addition of carbon and nitrogen sources to the medium did not affect, and even inhibited under certain conditions, the growth of LEM. The maximum temperature for the growth of LEM was around $25^{\circ}C$. Also, it grows better when agitated by shaking at 100 rpm for airration. The appropriate concentration of the extract to use was 10%. Under these conditions, LEM could reach to the confluency after cultivation of 12 days. Our extract formula seems better than other available media for LEM growth, producing higher crude protein content and better taste.
Kim, Yu-Jin;Jang, Seo-Jung;Park, Jung-Min;Kim, Chang-Uk;Park, Young-Seo
Food Engineering Progress
/
v.14
no.1
/
pp.65-74
/
2010
Culture conditions of L. plantarum TJ-LP-002, the garlic resistant strain isolated from pakimchi (green onion kimchi), were investigated for the use of feed additives. Acetic acid, citric acid, lactic acid, and tartaric acid were detected in the culture supernatant, and especially the concentrations of lactic acid and acetic acid significantly increased during cultivation. The antimicrobial activity of L. plantarum TJ-LP-002 was not affected by proteases, calatase or cellulase, which showed that the antimicrobial activity might be due to the production of acids rather than proteinaceous antimicrobial substances. L. plantarum TJ-LP-002 was resistant to neomycin sulfate, spectinomycin dihydrochloride, and lincomycin hydrochloride, sensitive to streptomycin sulfate, and intermediate resistant to ampicillin trihydrate, chloramphenicol, erythromycin, tetracycline hydrochloride, and kanamycin sulfate. The optimum initial pH of medium, fermentation temperature and time for the cell growth and antibacterial activity were pH 7.0, 30${^{\circ}C}$ and 24hr, respectively. The optimal composition of culture medium for the cell growth and antimicrobial activity was 3%(w/v) glucose as a carbon source, 3%(w/v) yeast extract as a nitrogen source, and manganese sulfate and ammonium citrate as inorganic salts. The combinatorial supplementation of these inorganic salts, rather than sole addition as an inorganic salt, resulted in better antibacterial activity.
From a sample of bamboo byproduct, the protease-producing yeast strain CO-1 was newly isolated. Strain CO-1 is spherical to ovoid in shape and measures $3.1-4.0{\times}3.8-4.4{\mu}m$. For the growth of strain CO-1, the optimal temperature and initial pH were $30^{\circ}C$ and 4.0, respectively. The strain was able to grow in 0.0-15.0%(w/v) NaCl and 0.0-9.0%(v/v) ethanol. Based on a phylogenetic analysis of its 18S rDNA sequences, strain CO-1 was identified as Pichia anomala. The extracellular protease produced by P. anomala CO-1 was partially purified by ammonium sulfate precipitation, which resulted in a 14.6-fold purification and a yield of 7.2%. The molecular mass of the protease was recorded as approximately 30 kDa via zymogram. The protease activity reached its maximum when 1.0%(w/v) CMC was used as the carbon source, 1.0%(w/v) yeast extract was used as the nitrogen source, and 0.3%(w/v) $MnSO_4$ was used as the mineral source. The protease revealed the highest activity at pH 7.0 and $30^{\circ}C$. This enzyme maintained more than 75% of its stability at a pH range of 4.0-10.0. After heating at $65^{\circ}C$ for 1 hr, the neutral protease registered at 60% of its original activity. The protease production coincided with growth and attained a maximal level during the post-exponential phase.
Ha, Gwangsu;Shin, Su-Jin;Jeong, Seong-Yeop;Yang, HoYeon;Im, Sua;Heo, JuHee;Yang, Hee-Jong;Jeong, Do-Youn
Journal of Life Science
/
v.29
no.8
/
pp.861-870
/
2019
This study was undertaken to establish optimum medium compositions for cost-effective mannitol production by Leuconostoc mesenteroides SRCM201425 isolated from kimchi. L. mesenteroides SRCM21425 from kimchi was selected for efficient mannitol production based on fructose analysis and identified by its 16S rRNA gene sequence, as well as by carbohydrate fermentation pattern analysis. To enhance mannitol production by L. mesenteroides SRCM201425, the effects of carbon, nitrogen, and mineral sources on mannitol production were first determined using Plackett-Burman design (PBD). The effects of 11 variables on mannitol production were investigated of which three variables, fructose, sucrose, and peptone, were selected. In the second step, each concentration of fructose, sucrose, and peptone was optimized using a central composite design (CCD) and response surface analysis. The predicted concentrations of fructose, sucrose, and peptone were 38.68 g/l, 30 g/l, and 39.67 g/l, respectively. The mathematical response model was reliable, with a coefficient of determination of $R^2=0.9185$. Mannitol production increased 20-fold as compared with the MRS medium, corresponding to a mannitol yield 97.46% when compared to MRS supplemented with 100 g/l of fructose in flask system. Furthermore, the production in the optimized medium was cost-effective. The findings of this study can be expected to be useful in biological production for catalytic hydrogenation causing byproduct and additional production costs.
To study the productivity of single cell protein from the petroleum hydrocarbon utilizing yeasts, 242 soil samples, such as oil soaked soil of gas stations and garage, coal, farm soil, and sewage, from 135 places in Korea were collected. From these samples 468 yeast strains which utilize petroleum hydrocarbon as a sole organic carbon source were isolated and identified by observing the growth rates. For the identified strains optimum culture conditions were determined and analysis of cell components were performed. 1. 90.8% of petroleum hydrocarbon utilizing yeast strains were found from oil soaked soil and about 10% from coal, farm soil and sewage etc. 2. The yeast strain of the highest cell productivity was isolated from oil soaked soil and was identified as Candida curvata HY-69-19. 3. The optimum culture conditions for the selected yeast strain were found to be pH 5.0, $28^{\circ}C$ and affluent aerated state. 4. Candida curvata HY-69-19 was found to utilize favorably the heavy gas oil fractionated at above $268.9^{\circ}C$ as carbon source and urea as inorganic nitrogen source. 5. The growth curve of this strain on heavy gas oil medium showed that the yeast has a lag phase up to 18 hours and logarithmic growth phase between 24 to 42 hours. Generation time was found to be between 3.8 and 4.5 hours during the logarithmic growth phase. 6. About 300 mg dried cells per heavy gas oil was harvested under the culture conditions of adjusted pH to 5.0 at time intervals of 6 hours for 54 hours and heavy gas oil urea for shaking culture medium. 7. Chemical composition of the yeast cell was found to be 40.25%, 14.81%, 24.32% and 10.63% for crude protein, crude lipid, carbohydrate and ashes, respectively.
To solve the problems with excessive accumulation of soil inorganic N and resulting saline soils from overuse of nitrogen fertilizer, the effect of sucrose application on decrease of soil inorganic N content and electrical conductivity (EC) was studied. Sucrose treatment greatly reduced ${NH_4}^+$-N content in soil. The amount of reduction was greater as the amount of sucrose treatment was increased. When ${NH_4}^+$-N content was reached the lowest point (about 10 mg $kg^{-1}$or lower), the C/N ratio, which determines the amount of sucrose treatment, was around 10 regardless of initial ${NH_4}^+$-N content. For the rate of ${NH_4}^+$-N reduction 15~36 hours was required to reduce the initial ${NH_4}^+$-N content to half, and 36~69 hours to lower ${NH_4}^+$-N content to the lowest point (about 10 mg $kg^{-1}$or lower). In addition, sucrose treatment greatly lowered ${NO_3}^-$-N content. In case of C/N ratio above 10, initial ${NO_3}^-$-N content of 348 mg $kg^{-1}$ was reduced to the lowest of 14~21 mg $kg^{-1}$. As for the rate of ${NO_3}^-$-N reduction by sucrose treatment, it took 36~60 hours for ${NO_3}^-$-N content to reach the lowest point for C/N ratio of 10 or higher, and it took 3 weeks, comparably longer time, for C/N ratio of 5. Lowering soil EC from sucrose treatment showed the same trend as ${NO_3}^-$-N content. As an important energy and carbon source for humankind, sugar should not be wasted and must be carefully applied to soil. In principle, the best way of preventing salt accumulation in soil is to optimize the fertilizer input. However, when over-fertilization should be dealt with, the sucrose treatment would be a possible and effective counter-measure to reduce overdosed nitrogen sources in soil.
Cordyceps pruinosa grows upon dead pupae of Lepidoptera and produces one or $3{\sim}4$ club-shaped stromata per host. The stromata have distinct club-shaped head and long stalk. The length of stromata varies from $1{\sim}3\;cm$. Apical head consists of densely crowded semi-immersed perithecia, which are $360{\sim}400\;{\times}\;180{\sim}200\;{\mu}m$ in size. Asci are $150\;{\mu}m$ in length and $2.8{\sim}3\;{\mu}m$ in diameter. Ascospores, which are $124{\sim}141\;{\mu}m$ in length, have thin thread-like structures in the middle with part-spores attached on both sides. Each ascospore does not separate into part-spores after dispersal, but each part-spore germinates and together develops a colony. The imperfect form produces phialides of $15{\sim}24\;{\times}\;2{\sim}3\;{\mu}m$ size, with spherical or spindle shaped conidia of $4{\sim}6\;{\times}\;1.8{\sim}2.4\;{\mu}m$ size, The anamorph was identified as Mariannaea elegans Samson. YMA and SDAY agar media with pH 7 was produced abundant mycelial growth with high density. Best mycelial growth was observed when dextrin was used as a carbon source. Lactose, saccharose and sucrose also produced high mycelial growth. Peptone, yeast extract and tryptone produced abundant mycelial growth, when used as nitrogen sources. Highest mycelial growth and density was observed when C/N ratio was 1 : 1 at the concentration of 12.5 g/l each. $KH_2PO_4$ was the best mineral source for mycelial growth. Highest mycelial dry wt. was produced in YM and SDAY broths. Optimum inoculum for 100 ml of liquid broth was 6 mycelial discs. Similarly, optimum liquid culture period was 7 days.
1. Isolation and identification of amylase-producing bacteria. The powerful strain A-12 and S-8 were respectively isolated from air and soil after screening a large number of amylase-producing bacteria. Their bacterial characteristics have been investigated and it has been found that all characteristics of strain A-12 and S-8 are similar to Bac. subtilis of Bergey's manual except for the acid formation from a few carbohydrates and the citrate utilization, i.e., the strain A-12 shows negative in the citrate utilization, and the acid formation from arabinose and xylose, S-8 shows negative in the acid formation from xylose. 2. Amylase production by Liquid cultures with solid materials. Several conditions for amylase production by strain A-12 in stationary cultures have been studied. The results obtained are as follows. (1) The optimum conditions are:temperature $35^{\circ}C$, initial pH 6.5 to 7.0 and incubation time 3 to 4 days. (2) The amylase production is not affected by the preservation period of the stock cultures. (3) Among the various solid material, the defatted soy bean is found to be the best for t1e amylase production. However, the alkali treatment of the defatted soy bean gives no effect contrary to the cage of defatted rape seed. The addition of soluble starch to the alkali extract of defatted soy bean shows the increased amylase production. (4) Up to 1% addition of ethanol to carbon dificient media gives the improved amylase production, whereas the above effect is not found in the case of carbon rich media. (5) The amylase production can be increased 2.5 times when 10% of defatted soy bean is admixed to cheaply available wheat bran. (6) The excellent effect is found for amylase production when 20% of wheat bran is admixed to defatted dry milk which is a poor medium. The activity is found to be $D^{40^{\circ}}_{30'}$ 7,000(L.S.V. 1,800) in 10% medium. (7) No significant effect is observed due to the addition of various inorganic salts. 3. Amylase production by solid cultures. Several conditions for amylase production by strain A-12 in wheat bran cultures have been studied and the results obtained are as follows. (1) The optimum conditions: are temperature $33^{\circ}C$, incubation lime 2 days, water content added 150 to 175% and the thickness of the medium 1.5cm, The activity is found to be $D^{40^{\circ}}_{30'}$ 36,000(L.S.V. 15,000) (2) No significant effect is found in the case of the additions of various organic and inorganic substances.
Burkholderia sp. D5, a polyaromatic hydrocarbons(PAHs)-degrading bacterium, was isolated from oil-contaminated soil. The bacterium could utilize phenanthrene (Phe) as a sole carbon source but could not use pyrene (Pyr). However, the strain could degrade Pyr when a cosubstrate such as yeast extract (YE) was supplemented. The PAH degradation rate of the bacterium was enhanced by the addition of other organic materials such as YE, peptone and glucose. YE was a particularly effective additive in stimulating cell growth as well as PAH degradation. When 1 g-YE/L was supplemented into the basal salt medium (BSM) with 215 mg-Phe/L, the specific growth rate (0.28 h-1) and Phe-degrading rate (29.30 μmol/L/h) were enhanced approximately ten and two times more than those obtained in the BSM with 215 mg-Phe/L, respectively. Through kinetic analysis, the maximum specific growth rate (μmax) and PAH degrading rate (Vmax) for Phe were obtained as 0.34/h and 289 ${\mu}mol$/L/h, respectively. Also, μmax and Vmax for Pyr were 0.27 h-1 and 50 ${\mu}mol$/L/h, respectively. The degradation rates for each Phe (2.20 μmol/L/h) and Pyr (2.18 μmol/L/h) were lower in mixture substrates than in a single substrate (29.30 ${\mu}mol$/L/h and 9.58 ${\mu}mol$/L/h, respectively). Burkholderia sp. D5 can degrade Phe and Pyr contained in soil, and the PAH degradation rates in soil were 20.03 ${\mu}mol$/L/h for Phe and 1.09 ${\mu}mol$/L/h for Pyr.
A series of experiments was conducted to study the behavior of the phosphorus added to the soils having the high phorphorus fixing capacity derived from volcanic ash in Cheju Island. Soil samples were taken from different depths of 0-10, 10-30, and 30-50cm in six citrus orchards where heavy application of phosphate fertilizer has been practised. Various forms of phosphorus were determined and phosphorus adsorption experiments were performed. The results obtained can be summarized as follows: 1. The content of inorganic phosphorus fractions determined by the method of Chang and Jackson was: water soluble P
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