• Biotechnology and Bioprocess Engineering:BBE
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• 제8권3호
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• pp.187-191
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• 2003

Proteolytic enzymes existing in plant cell cultured media are the major reason for the loss of secreted human granulocyte-macrophage colony-stimulating factor (hGM-CSF). The addition of pepstatin, aprotinin and PMSF relatively decreased the proteolytic degradation of hGM-CSF in a conditioned medium, but sufficient prevention against the proteolytic activity could not be obtained with chemical protease inhibitors. Gelatin, as a competitive substrate for protease, showed a stabilizing effect in a conditioned medium. Compared to the initial hGM-CSF concentration in a conditioned medium. with 10 g/L of gelatin, 68% of the hGM-CSF remained after 5 days. In a cell culture experiment, 5 g/L of gelatin significantly stimulated the hGM-CSF production and accumulation in culture media, with no growth inhibition. compared to the controls (4.72 $\mu\textrm{g}$/L), the extracellular hGM-CSF level could be increased to 39.78 $\mu\textrm{g}$/L with the addition of 5 g/L of gelatin.

• Plant Resources
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• 제2권1호
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• pp.42-48
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• 1999

This study was undertaken to examine the feasibility of black locust (Robinia pseudoacacia) as substrates for several edible mushrooms. For the cultivation of several edible and/or medicinal fungi on black locust, optimum bulk densities, synthetic or semisynthetic additives, natural additives and pretreatment methods were investigated. Fruit body yields of the fungi on various sawdust media composed of different wood species were also analyzed for testing the capability of black locust as a substrate for mushroom production. Mycelial growths decreased proportional when the bulk density increased. The most suitable carbon and nitrogen sources as additives to promote the mycelial growth were sucrose (2%, w/w) and ammonium phosphate (0.2%, w/w) respectively. When corn-powder and beer-waste as natural additives were added to sawdust of black locust showed the significant growth of mycelia. And the optimum mixing ratio was 10:2:1 (sawdust: corn-powder: beer-waste, w/w). Black locust after cold water treatment showed the outstanding mycelial growths. Any significant changes of pH, moisture content (%) and dry-weight losses (%) could not be found among culture substrates (sawdust of black locust, oak and poplar wood) examined before and after harvesting of fruit bodies. Yield of fruit bodies on black locust culture media were comparable with those culture media composed with oak and poplar wood. The present work indicated strongly the potentiality of black locust as raw materials for edible and medicinal mushrooms.

• KSBB Journal
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• 제11권3호
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• pp.367-373
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• 1996

Rhodotorula sp. Y -55 효모의 중식특성을 etha nol, acetic acid 또는 acetaldehyde 상에서 batch culture로 조사하였다. 기질농도가 중가함에 따라 중식수율이 감소하고 유도기가 연장되었다. 비성장속도(${\mu}$)는 지수기에서 ethanol과 acetaldehyde의 초기농도에 따라 변화하였으나 acetic acid는 이와 상관이 없었으며 그 값은 $O.107h^{-1}$로 나타났다. 최대 비성장속도는 20g/L의 ethanol에서 $O.270h^{-1}$, O.2g/L의 acetaldehyde에서 $O.041h^{-1}$이었다. 또 최대 중식수융은 109/L의 ethanol에서 32.6%, 20g/L의 a acetic acid에서 25.6%와 O.2g/L의 acetaldehyde 에서 45%였다. 세포내 조단백질과 핵산 함량은 e­t thanol상에서 각각 41.5wt%와 4.9wt %였고 acetic acid상에서는 각각 40.2wt %와 4.7wt%로 나타났다.

• Journal of the Korean Wood Science and Technology
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• 제17권3호
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• pp.52-60
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• 1989

This study was carried out to know the possibility of simultaneous utilization of laccase from white-rot fungus with cellulase on enzymatic hydrolysis of cellulosic substrate from autohydrolyzed oak wood. Laccases from 3 white-rot fungi, Pleurotus ostreatus. Ganoderma lucidum, and Phanerochaete chrysosporium, were isolated, purified and measured their activities. The highest activity was shown in Pleurotus ostreatus and the lowest in Phanerochaete chrysosporium. Laccase from Pleurotus ostreatus has optimum pH of 5.94, Km value of 3.209 mM and appeared to be stable at relatively wide pH range, 4.7-8.72. Temperature stability showed that 60% activity was preserved after 40 minutes at $50^{\circ}C$. Laccase from Ganoderma lucidum reached to the maximum activity during 15-20 day incubation. This enzyme has optimum pH of 6.45, Km value of 6.71 mM and pH range of 5.0-9.0 for stabilization. 95% activity was preserved at $30^{\circ}C$ and 58% activity at $50^{\circ}C$. Concerned to the enzymatic hydrolysis of cellulosic substrate with both enzymes, cellulase and laccase, simultaneously, mixed culture filtrates and mycellium extracts were shown higher hydrolysis rates than those of Trichoderma viride. There were no significant differences in the extent of hydrolysis among various mixed culture filtrates and mycellium extracts.

• 한국근적외분광분석학회:학술대회논문집
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• 한국근적외분광분석학회 2001년도 NIR-2001
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• pp.1157-1157
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• 2001

Yeast, Kurtzurnanomyces sp. I-11, produces biosurfactant, mannosyl erythritol lipid (MEL), from soya oil. The properties of biosurfactant MEL include low-toxicity and high biodegradability. MEL provides new possibilities for a wide range of industrial applications, especially food, cosmetic, pharmaceutical fields and chemicals for biotechnology. In the fermentation process, techniques of measuring and controlling substrates and products are important to obtain high productivity with optimum concentrations of substrate and product in the culture broth. The measurement system for the concentrations of soya oil and MEL in the fermentation process was developed using near-infrared spectroscopy (NIRS). Soya oil and MEL in the culture broth were extracted with ethyl acetate and NIR spectra was carried out between the second derivative NIR spectral data at 1312 and 2040 nm and MEL concentrations obtained using a thin-layer chromatography with a flame-ionization detector (TLC/FID) method. A calibration equation for soya oil was results of the validation of the calibration equation, good agreement was observed between the results of the TLD/FID method and those of the NIRS method for both constituents. NIR method was applied to the measurement of the concentrations of MEL and soya oil in the practical fermentation and good results were obtained. The study indicates that NIRS is a useful method for measurement of the substrate and product in the glycolipid fermentation.

• Korean Chemical Engineering Research
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• 제52권3호
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• pp.302-306
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• 2014

목질계 분해효소 활성 증대를 위해 밀짚을 이용한 고체발효에서 주요 발효인자의 최적화를 수행하였다. Trichoderma reesei와 Aspergillus niger를 이용한 혼합배양에서 고체발효에 주요한 영향을 미친다고 알려진 배양온도, pH, 수분함량과 고체기질 크기를 순차적 최적화를 진행하였다. 실험에 적용 된 인자 모두 목질계 분해효소 활성에 유의한 효과를 주었으며, 발효온도 $40^{\circ}C$, pH 7, 수분함량 75%와 고체기질 크기 0.25~0.5 mm가 목질계 분해효소 생산을 위한 최적 조건임을 알 수 있었다. 최적조건 하에서 밀짚을 이용한 고체발효를 수행하였을 때, 효소활성 기준 cellulase 10.3 IU, endoglucanase 100.3 IU, ${\beta}$-glucosidase 22.9 IU와 xylanase 2261.7 IU/g dry material을 배양 96시간에 확인할 수 있었다. 본 결과는 기존 효소활성 대비 각각 72.6, 48.7, 55.2와 51.9% 증가한 수치로 혼합배양과 순차적 최적화를 적용하여 효과적인 목질계 분해효소 활성 증대가 가능함을 확인하였다.

• 한국수소및신에너지학회논문집
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• 제22권4호
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• pp.451-457
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• 2011

Purple non-sulfur (PNS) bacterium $Rhodobacter$ $sphaeroides$ KD131 was studied with the aim of achieving maximum hydrogen production using various carbon and nitrogen sources at different pH conditions. Cells grew well and produced hydrogen using $(NH_4){_2}SO_4$ or glutamate as a nitrogen source in combination with a carbon substrate, succinate or malate. During 48h of photo-heterotrophic fermentation under 110$W/m^2$ illumination using a halogen lamp at $30^{\circ}C$, 67% of 30mM succinate added was degraded and the hydrogen yield was estimated as 3.29mol $H^2$/mol-succinate. However, less than 30% of formate was consumed and hydrogen was not produced due to a lack of genes coding for the formate-hydrogen lyase complex of strain KD131. Initial cell concentrations of more than 0.6g dry cell weight/L-culture broth were not favorable for hydrogen evolution by cell aggregation, thus leading to substrate and light unavailability. In a modified Sistrom's medium containing 30mM succinate with a carbon to nitrogen ratio of 12.85 (w/w), glutamate produced 1.40-fold more hydrogen compared to ammonium sulfate during the first 48h. However, ammonium sulfate was 1.78-fold more effective for extended cultivation of 96h. An initial pH range from 6.0 to 9.0 influenced cell growth and hydrogen production, and maintenance of pH 7.5 during photofermentation led to the increased hydrogen yield.

• 한국미생물·생명공학회지
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• 제13권3호
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• pp.239-243
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• 1985

Ethanol, Acetic acid 및 Acetaldehyde를 단일 탄소원으로 하여 Candida sP. JY-5 효모의 증식특성을 batch culture를 통해 검토하였다. Ethanol상에서 이 효모의 지수적 증식기 동안의 비증식도 ($\mu$) 값은 농도가 0.5g/l 이상일 경우 그 초기 농도에 따라 변화하나 비례하지는 않았다. 최대 비증식도는 0.291$hr^{-1}$이었고 최대 증식수율은 10g/l 농도에서 61.2 %이었다. Acetic acid상에서의 비증식도는 이 기질의 초기 농도에 관계없이 일정하였으며 그 값이 0.106$hr^{-1}$. 최대 증식수율은 10g/l 농도에서 46.8%로 나타났다. 한편 Acetaldehyde 기질에서 지수적 증식기 동안의 최대 비증식도는 0. 063$hr^{-1}$이었고 최대 증식수율은 0.2g/l 농도에서 44.9%였다.

• 한국균학회소식:학술대회논문집
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• 한국균학회 2018년도 춘계학술대회 및 임시총회
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• pp.29-29
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• 2018

Sparassis latifolia is called "Cauliflower Mushroom" and is known as an edible mushroom that has high content of ${\beta}$-glucan. Recently, artificial cultivation of S. latifolia has been done by bottle, plastic bag and wood cultivation in Korea. However it is not widely used because there are low incubation ratio and yield. For the high efficiency of production, we aim to find the superior strains and media for better mycelial and fruit body growth. First, we analyzed the genetic relationship among 31 strains and divided five groups with three kinds of URP primers. And then ten strains were selected from five groups based on the experiment of mycelial growth. The suitability of media for mycelial growth was different according to media type. The suitable solid and liquid media for mycelial growth of S. latifolia isolates were PDA and M2, respectively. In addition, with regard to C/N ratio, the mycelial growth increased even until C/N 160. Second, we investigated the production of fruitbody of the strains by plastic bag cultivation. The substrate was mixed with larch sawdust, corn flour, and wheat flour (8:1:1, v/v). Moisture content of substrate was controlled by about 60% with 10% molasses solution. Out of 31 strains, 19 strains formed primordia. The eight strains produced more than 140g/1kg in fresh weight. Third, molasses culture media was selected for the mycelial growth. And molasses suitable sugar content and input aeration were around 8Brix% and 0.3~0.6vvm, respectively. The longer the incubation period is, the more dried weight of mycelia increased, but medium volume decreased. Therefore, the best incubation period was 9 to 11 days depending on strains. In the future, research project entitled development of culture system and new variety for stable production of S. latifolia will be considered as a new item.

• Journal of Acupuncture Research
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• 제17권3호
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• pp.176-187
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• 2000

Objectives : This study was purposed to investigate the antioxidative effects of Paeoniae radix aqua-acupuncture solution(PR) on culture liver cell system, lipid peroxidation and antioxidative enzyme activities in tert-butyl hydroperoxide(t-BHP) treatmented conditions. Methods : Cultured normal rat liver cell(Ac2F) were prepared and incubated with or without PR(at 2% volume in culture medium). After 16~18hr, cells placed in DMEM medium without serum, and then incubated with 1mM t-BHP for 2hr. Viable cells were detected by MTT assay, and the levels of lipid peroxide(LPO) were measured by TBA method. And catalase activity was measured as the decrease in hydrogen peroxide absorbance at 240nm on spectrophotometer using 30mM hydrogen peroxide. Superoxide dismutase(SOD) were assayed by recording the inhibition of nitro blue tetrazolium reduction with xanthine and xanthine oxidase. Glutathione peroxidase(GPX) activity was determined by the modified coupled assay developed by Paglia and Lawrence. The reaction was started by addition of 2.2mM hydrogen peroxide as substrate. The change in absorbance at 340nm was measured for 1min on spectrophotometer. Glutathione-S-transferase(GST) activity was assayed with CDNB as substrate and enzyme activity of GST towards the glutathione conjugation of CDNB. Results : Cell killing was significantly enhanced by addition of t-BHP compared to those of untreated group. PR pretreated cell resisted the toxic effects of t-BHP. LPO levels of t-BHP treatment group were significantly higher than other groups. This increased level was significandy reduced by PR pretreatment. The t-BHP treatment resulted in a decrease of catalase, GPX and GST activities. By contrast, PR pretreatment markedly increased compare to those of untreated groups. Conclusions : T-BHP which can produce intracellular free radical was used for inducer of the peroxidation of cellular lipids. PR protected the cell death induced by t-BHP and significantly increased cell viabiliry in the normal rat liver cell, and showed effective inhibition of lipid peroxidation, and elevations of catalase, GPX and GST activities. These results suggested that PR might play a protective role in lipid peroxidation by free radicals.