• Title/Summary/Keyword: yeast protein

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Construction of High Sensitive Detection System for Endocrine Disruptors with Yeast n-Alkane-assimilating Yarrowia lipolytica

  • Cho, Eun-Min;Lee, Haeng-Seog;Eom, Chi-Yong;Ohta, Akinori
    • Journal of Microbiology and Biotechnology
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    • v.20 no.11
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    • pp.1563-1570
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    • 2010
  • To construct a highly sensitive detection system for endocrine disruptors (EDs), we have compared the activity of promoters with the n-alkane-inducible cytochrome P450 gene (ALK1), isocitrate lyase gene (ICL1), ribosomal protein S7 gene (RPS7), and the translation elongation factor-1${\alpha}$ gene (TEF1) for the heterologous gene in Yarrowia lipolytica. The promoters were introduced into the upstream of the lacZ or hERa reporter genes, respectively, and the activity was evaluated by ${\beta}$-galactosidase assay for lacZ and Western blot analysis for hER${\alpha}$. The expression analysis revealed that the ALK1 and ICL1 promoters were induced by n-decane and by EtOH, respectively. The constitutive promoter of RPS7 and TEF1 showed mostly a high level of expression in the presence of glucose and glycerol, respectively. In particular, the TEF1 promoter showed the highest ${\beta}$-galactosidase activity and a significant signal by Western blotting with the anti-estrogen receptor, compared with the other promoters. Moreover, the detection system was constructed with promoters linked to the upstream of the expression vector for the hER${\alpha}$ gene transformed into the Y. lipolytica with a chromosome-integrated lacZ reporter gene under the control of estrogen response elements (EREs). It was indicated that a combination of pTEF1p-hER${\alpha}$ and CXAU1-2XERE was the most effective system for the $E_2$-dependent induction of the ${\beta}$-galactosidase activity. This system showed the highest ${\beta}$-galactosidase activity at $10^{-6}\;M\;E_2$, and the activity could be detected at even the concentration of $10^{-10}\;M\;E_2$. As a result, we have constructed a strongly sensitive detection system with Y. lipolitica to evaluate recognized/suspected ED chemicals, such as natural/synthetic hormones, pesticides, and commercial chemicals. The results demonstrate the utility, sensitivity, and reproducibility of the system for identifying and characterizing environmental estrogens.

Molecular Cloning and Characterization of Salt-inducible Aldolase from Salicornia herbacea (퉁퉁마디로부터 염에 의하여 유도되는 Aldolase 유전자의 분리 및 발현분석)

  • Cha, Joon-Yung;Netty Ermawati;Kim, Soon-Gil;Lee, Jeung-Joo;Lim, Chae-Oh;Chung, Woo-Sik;Lee, Kon-Ho;Son, Dae-Young
    • Journal of Plant Biotechnology
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    • v.30 no.4
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    • pp.323-328
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    • 2003
  • Soil salinity is one of the most serious abiotic stresses limiting the productivity of agricultural crops. To cope with salt stress, plants respond with physiological, developmental and biochemical changes, including the synthesis of a number of proteins and the induction of gene expression. Salicornia herbacea is a halophytic plant that grows in salt marshes and on muddy seashores. In order to understand the biochemical and molecular mechanisms of salt tolerance in S. herbacea, we isolated several genes that involved in the salt tolerance by mRNA differential display. Here we report the cloning of a cDNA encoding fructose-1, 6-bisphosphate aldolase, named ShADL, which is 1293 bp long and contains an open reading frame consisted of 359 amino acids with calculated molecular mass of 39 kDa. ShADL protein showed 86% identity with Arabidopsis and 78% with aldolase of common ice plant. Northern blot analysis revealed that the transcript of ShADL gene was increased dramatically depending on the NaCl concentrations.

Effects of Roasted Cassia tora L. Extracts on the Chemical Changes and Microbial Growth (열처리가 결명자의 화학성분 변화 및 추출물의 균체증식에 미치는 영향)

  • Yun, Jong-Bum;Kim, Kyeong-Gu;Sa, Tong-Min;Lee, Young-Tack;Joo, Hyun-Kyu
    • Applied Biological Chemistry
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    • v.40 no.6
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    • pp.472-477
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    • 1997
  • The effects of roasting Cassia tora L. were investigated for proximate composition, color, volatile flavor profile, microbial growth and alcohol fermentation. While moisture, protein and fat contents decreased with increasing roasting temperature, fiber and ash contents increased. The L, a and b values of Cassia tora L. extracts decreased with increasing temperature, and only a small difference in total color difference$({\Delta}E)$ was observed. A little difference in major flavor components between raw and roasted treatment was found during roasting. Furfuryl alcohol, a major component of coffee flavor, was separated from Cassia tora L. extracts extracted with ethyl ether. The yeast growth was the highest on the water-extract of Cassia tora L. roasted at $160^{\circ}C$. With increased levels of water-extract at $160^{\circ}C$, S. cerevisiae grew rapidly for 24 hr incubation and the growth rate was higher than the unroasted control group. The growth rate of Bacillus subtilis was the highest in a treatment of 0.5% concentration. Little differences in ${\alpha}-amylase$ produced from Bacillus subtilis were observed among the treatment groups. The total alcohol content increased with increasing roasting temperature during alcohol fermentation.

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Quality Characteristics of Ginseng Treated by Hot Air Drying after Being Dried using Super-heated Steam (초급속 과열증기를 이용한 인삼의 품질 특성)

  • Kim, Kyung-Tack;Hong, Hee-Do;Kim, Sung-Soo
    • Journal of Ginseng Research
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    • v.33 no.4
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    • pp.361-366
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    • 2009
  • This research aims to develop a new drying technology using super-heated steam to reduce the time required for drying ginseng and to enhance its quality. The drying rate of ginseng powder treated with hot-air drying after drying with super-heated steam was about 40% faster than the drying rate of ginseng powder treated with hot-air drying. Minimal difference was found between hot-air drying and hot-air drying after drying with super-heated steam in the general composition of the ginseng powder, such as its crude lipid, crude protein, crude fiber, and ash. Moreover, no difference was found between hot-air drying and hot-air drying after drying with super-heated steam in terms of the ginseng powder's mineral and free sugar contents. The crude saponin contents was 5.43% in the ginseng treated with hot-air drying and 5.4% in the ginseng treated with hot-air drying after drying with super-heated steam. The ginseng powder that was treated with hot-air drying after drying with super-heated steam had a slightly higher redness compared to the ginseng powder that was treated with hot-air drying alone. The number of microorganisms of the ginseng powder treated with hotair drying after drying with super-heated steam showed a reduction of one log cycle in the total bacteria and two log cycles in the yeast molds compared to the ginseng powder treated with hot-air drying alone.

Soju brewing characteristics of yeast strains N4 and N9 isolated from Korean traditional Nuruk (재래누룩에서 분리한 N4와 N9 효모의 증류식 소주 양조특성)

  • Choi, Han-Seok;Kang, Ji-Eun;Jeong, Seok-Tae;Kim, Chan-Woo;Baek, Seong-Yeol;Yeo, Soo-Hwan
    • Food Science and Preservation
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    • v.24 no.5
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    • pp.714-724
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    • 2017
  • Soju is a Korean traditional distilled alcoholic beverage produced from mashes various crops and Nuruk which is cultured with wild microorganisms. This study was conducted to investigate rice-Soju brewing characteristics of yeasts isolated from Korean traditional Nuruk. The general components of rice (Hanarumbyeo) raw materials were 14.7 g of water, 6.8 g of crude protein, 0.9 g of crude lipid, 0.4 g of crude ash, and 76.5 g of carbohydrate in 100 g. Saccharifying and proteolytic activities in Hanarumbyeo ipguk (solid-state culture of Aspergillus luchuensis) were also determined. The alcohol content of the fermented wash from isolates was 15.37-16.58% (v/v), which is 16.7-36.0% higher than that of industrial yeasts (12.33-13.19%). Reducing sugar contents were 2.04-3.92 and 7.92-8.78 g/100 mL in the isolates and industrial yeasts, respectively. The isolated yeasts showed 25.2-52.7% higher yield of distillates (41% alcohol) compared to industrial yeasts. Forty-one components were detected in the rice distillated Soju (25% alcohol) and principal component analysis revealed differences between the isolated and industrial yeasts with respect to the contents of i-BuOH, isobutanal diethyl acetal, ethyl caprate, and tetradecanoic acid.

Production of Single Cell Protein on Petroleum Hydrocarbon -IV. On the Continuous Fermentation and Some Cultivation Conditions for Candida tropicalis KIST 351- (석유탄화수소를 이용한 단세포단백질의 생산에 관한 연구 -IV. Candida tropicalis KIST 351의 연속배양 및 몇가지 조건에 관하여-)

  • Lee, Yong-Hyun;Pyun, Yoo-Ryang;Kwon, Tai-Wan
    • Korean Journal of Food Science and Technology
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    • v.4 no.3
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    • pp.200-205
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    • 1972
  • Effects of several different petroleum fractions (LGO, HGO, VGO, Diesel oil, SP(E), HGO-wax, L/M-wax), stepwise addition of calculated amounts of HGO at defined intervals, recycling of spent media on cell growth of Candida tropicalis KIST 351 were studied using $2.5{\ell}$ fermenter by batch process. In addition, continuous cultivation of the yeast was also performed in the light of biomass production using $28{\ell}$ fermenter with LGO. 1) Cell concentration, yield on the basis of gas oil and n-paraffin with the petroleum fractions were in the range of $11{\sim}15g/{\ell}$, $10{\sim}12%$ and $77{\sim}82%$, respectively. 2) By stepwise addition of the gas oil, cell concentration and yield on the oil were increased up to 18.9 g/land 13%, respectively. 3) Spent medium slowed emulsifying ability of hydrocarbon and stimulating effect on the cell growth. Without additional supplementation of $Mg^{++}$ up to 20% of spent medium could be reused, while by adding of the $Mg^{++}$, 50% of medium could be recycled. 4) Optimum condition of continuous cultivation for biomass production was attained at the dilution rate of $D=0.1{\sim}0.125\;hr^{-1}$. Maximum yield coefficient on consumed n-paraffin was 0.94 at $D=0.1\;hr^{-1}$, however, 24% of supplied n-paraffin in the media was not utilized at this dilution rate.

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Comparison of Physicochemical Properties between Organic and Conventional Soybean by Steaming Treatment (증자 처리한 일반콩과 유기콩의 이화학적 특성 분석)

  • Kim, Jin-Sook;Park, Soo-Jin;Choi, Mi-Kyeong;Moon, Eun-Young;Kang, Myung-Hwa
    • Journal of the East Asian Society of Dietary Life
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    • v.20 no.6
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    • pp.963-968
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    • 2010
  • The differences between organic and conventional soybean were studied based on their physicochemical properties by steaming treatment. There was no difference between the two kinds of soybean regarding crude protein, crude fat and crude ash contents. Reduction in sugar content was higher in raw organic soybean than raw conventional soybean. However, there was no difference between the two soybeans after steaming. Amino nitrogen content was higher in organic than conventional soybean. Further, there was no difference between the two soybeans after steaming. Regarding pH, acidity and salinity, there was no differences between the two soybean after steaming. Organic soybean was more bright than conventional soybean after steaming. The yeafercontent and number of bacteria in raw soybean were not different between the two soybean types. The number of fungi was higher in organic than conventional soybean. Upon steaming, the yeast number was different between two kinds of soybean. Further, the number of bacteria was higher in organic than conventional soybean. The number of fungi was not different between the two soybeans.

Production of Keratinolytic Protease by Bacillus pumilus RS7 and Feather Hydrolysate As a Source of Amino Acids (Bacillus pumilus RS7에 의한 난분해성 케라틴 분해효소의 생산 및 아미노산 공급원으로서 우모 분해산물)

  • Woo, Eun-Ok;Kim, Min-Ju;Son, Hyeng-Sik;Ryu, Eun-Youn;Jeong, Seong-Yun;Son, Hong-Joo;Lee, Sang-Joon;Park, Geun-Tae
    • Journal of Environmental Science International
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    • v.16 no.10
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    • pp.1203-1208
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    • 2007
  • Feathers are produced in huge quantities as a waste product at commercial poultry processing plants. Since feathers are almost pure keratin protein, feather wastes represent an alternative to more expensive dietary ingredients for animal feedstuffs. Generally they become feather meal used as animal feed after undergoing physical and chemical treatments. These processes require significant energy and also cause environmental pollutions. Therefore, biodegradation of feather by microorganisms represents an alternative method to prevent environment contamination. The aim of this study was to investigate cultural conditions affecting keratinolytic protease production by Bacillus pumilus RS7. We also assessed the nutritive value of microbial and alkaline feather hydrolysates, The composition of optimal medium for the keratinolytic protease was fructose 0.05%, yeast extract 0.3%, NaCl 0.05%, K2HPO4 0.03%, KH2PO4 0.04% and MgCl2 6H2O 0.01%, respectively. The optimal temperature and initial pH was $30^{\circ}C$ and 9.0, respectively. The keratinolytic protease production under optimal condition reached a maximum after 18 h of cultivation. Total amino acid content of feather hydrolysates treated by NaOH and B. pumilius RS7 was $113.8\;{\mu}g/ml$ and $504.9\;{\mu}g/ml$, respectively. Essential amino acid content of feather hydrolysates treated by NaOH and B. pumilius RS7 was $47.2\;{\mu}g/ml$ and $334.0\;{\mu}g/ml$, respectively. Thus, feather hydrolysates have the potential for utilization as an ingredient in animal feed.

Characterization of a Novel DWD Protein that Participates in Heat Stress Response in Arabidopsis

  • Kim, Soon-Hee;Lee, Joon-Hyun;Seo, Kyoung-In;Ryu, Boyeong;Sung, Yongju;Chung, Taijoon;Deng, Xing Wang;Lee, Jae-Hoon
    • Molecules and Cells
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    • v.37 no.11
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    • pp.833-840
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    • 2014
  • Cullin4-RING ubiquitin ligase (CRL4) is a family of multi-subunit E3 ligases. To investigate the possible involvement of CRL4 in heat stress response, we screened T-DNA insertion mutants of putative CRL4 substrate receptors that exhibited altered patterns in response to heat stress. One of the mutants exhibited heat stress tolerance and was named heat stress tolerant DWD1 (htd1). Introduction of HTD1 gene into htd1-1 led to recovery of heat sensitivity to the wild type level, confirming that the decrease of HTD1 transcripts resulted in heat tolerance. Therefore, HTD1 plays a negative role in thermotolerance in Arabidopsis. Additionally, HTD1 directly interacted with DDB1a in yeast two-hybrid assays and associated with DDB1b in vivo, supporting that it could be a part of a CRL4 complex. Various heat-inducible genes such as HSP14.7, HSP21, At2g03020 and WRKY28 were hyper-induced in htd1-1, indicating that HTD1 could function as a negative regulator for the expression of such genes and that these genes might contribute to thermotolerance of htd1-1, at least in part. HTD1 was associated with HSP90-1, a crucial regulator of thermotolerance, in vivo, even though the decrease of HTD1 did not affect the accumulation pattern of HSP90-1 in Arabidopsis. These findings indicate that a negative role of HTD1 in thermotolerance might be achieved through its association with HSP90-1, possibly by disturbing the action of HSP90-1, not by the degradation of HSP90-1. This study will serve as an important step toward understanding of the functional connection between CRL4-mediated processes and plant heat stress signaling.

Characteristics of Kiwifruit-Added Traditional Kochujang (키위 첨가 전통고추장의 품질 특성)

  • Kim, Young-Soo;Song, Geun-Seoup
    • Korean Journal of Food Science and Technology
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    • v.34 no.6
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    • pp.1091-1097
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    • 2002
  • In order to improve palatability and quality of traditional kochujang, kiwifruit (Actinidia deliciosa) was added to traditional kochujang fermented for 3 months. Physicochemical and microbial characteristics were investigated during fermentation at $30^{\circ}C$. Moisture contents of all treated kochujangs increased with increasing amount of kiwifruit added during fermentation. pH of kiwifruit-added kochujang was lower than that of the control kochujang, whereas no significant differences in titratible acidity was observed among all treatments. Crude protein and salt contents of kiwifruit-added kochujang were lower than those of the control, whereas no difference in crude fat contents were found. Ethanol and amino-nitrogen contents of 6 and 9% kiwifruit-added kochujang were higher than those of the control. Fructose contents of kiwi-added kochujangs were higher than that of the control. Major organic acids of kochujang were in the order of malic acid > citric acid > succinic acid > acetic acid > lactic acid > oxalic acid. Succinic acid level increased significantly during fermentation and was higher in kiwifruit-added kochujang than in the control. Bacterial cell counts of all treatments were not different, and viable cell count of yeast was slightly higher in kiwifruit-added kochujang only at the initial fermentation period. Sensory evaluation revealed that the addition of 9% kiwifruit was the optimum condition for improving kochujang quality.