• Title/Summary/Keyword: KEX2

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Endoproteolytic Processing of Human Parathyroid Hormone in Saccharomyces cerevisiae Mutants Lacking Genes Encoding the Yap3, Mkc7, and Kex2 Endoproteases

  • Choi, Won-A.;Kang, Hyun-Ah;Chung, Bong-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.9 no.5
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    • pp.679-682
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    • 1999
  • When the human parathyroid hormone (hPTH) is expressed as a secretory product in S. cerevisiae, most of the secreted hPTH is internally cleaved by endoproteolytic processing. To investigate whether the yeast endoproteases such as Kex2p, Yap3p, and Mkc7p are involved in the endoproteolytic processing of hPTH in S. cerevisiae, hPTH was expressed in S. cerevisiae mutants deficient in one or two of the following well-known endoproteases such as Kex2p, Mkc7p, and Yap3p. Among these mutants, the yap3-disrupted(yap3$\Delta$) and yap3/mkc7-disrupted (yap3Δmkc7$\Delta$) yeasts showed a significant reduction in the extent of hPTH proteolysis. In contrast, the mkc7-disrupted (mkc7$\Delta$) yeast did not reduce the proteolysis of hPTH as compared to the wild type. This suggests that Mkc7p is not involved in the endoproteolytic processing of hPTH. It was also found that the kex2-disrupted (kex2$\Delta$) mutant was not able to secrete a detectable amount of hPTH.

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Expression of Carboxypeptidase Z cDNA from Absidia zychae in saccharomyces cerevisiae and its characteristics (Saccharomyces cerevisiae에서 Absidia zychae 의 Carboxypeptidase A cDNA 의 발현과 특성)

  • 이병로;김종화
    • Microbiology and Biotechnology Letters
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    • v.23 no.2
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    • pp.150-155
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    • 1995
  • Carboxypeptidase Z(CPZ) cDNA of Absidia zychae was experssed in Saccharomyces cerevisiae. The expressed CPZ(YCPZ) was secreted about 30 mg/l into the medium and has a little higher molecular weight than the wild type CPZ in SDS-PAGE. By the result of N-terminal amino acid sequencing, YCPZ has additional 15 amino acids residues in N-terminus of CPZ. But YCPZ shows no difference with CPZ in enzyme activity and substrate specificity. For the identification of processing mechanism of YCPZ, 36-Arg was changed to 36-Thr by site specific mutagenesis. Mutant YCPZ does not processed at 36-Thr. It was, therefore, concluded that the YCPZ was processed by KEX2. According to endo F treatment, high amount of carbohydrate was N-glycosylated in YCPZ.

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Potassium and Clay Minerals in Upland Soils (밭 토양(土壤)의 점토(粘土) 광물(鑛物)과 가리(加里))

  • Kim, Tai-Soon
    • Korean Journal of Soil Science and Fertilizer
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    • v.10 no.3
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    • pp.135-151
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    • 1977
  • The present paper summerizes the studies on clay mineralogical characteristics of Korean soil, relationship between potassium and clay minerals, potassium release pattern of clay minerals and utilization of clay minerals for soil conditioner and fertilizers, which have been carried out in this laboratory. 1. The red yellow podzolic soil is mostly abundant in the upland of Korea and mainly consists of halloysite and weathered intermediates of mica such as illite and vermiculite. 2. With regard to soil parent material, kaolin mineral occurs abundant in soils derived from granite and granite gneiss. Mica is dominant in basaltic soil. The main clay mineral of the soil, originated from the Tertiary, is found montmorillonite and the volcanic soil of Jeju Island has plenty of allophane as its main clay mineral. 3. It is confirmed that the soil fertility depends on the composition of clay minerals. The red yellow podzolic soil, containing lot of kaolin, shows low productivity while the montmorillonite soil has higher productivity. 4. The release rate of solid phase potassium (micas and fixed potassium) follows the 1st order reaction equation in the equilibrium solution of $IN-NH_4OAc$. The potassium release constant is positively correlated with the mica content of the clay but negatively correlated with the content of $14.5{\AA}$ minerals. On the other hand, the potassium release constant has very high correlation with the ratio(Kex/Kt) of exchangeable potassium(Kex) to total potassium(Kt). 5. It is also found that Kex/Kt has rather high correlation with the content of mica and $14.5{\AA}$ minerals existed in the clay as well as the mica content of the soil.

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TREATMENT OF PHENOL CONTAINED IN WASTE WATER USING THE HETEROGENIZED FENTON SYSTEM

  • Kim, Seong-Bo
    • Environmental Engineering Research
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    • v.12 no.1
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    • pp.30-35
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    • 2007
  • Fenton system using homogeneous iron catalyst is very powerful in the degradation of organic compounds, but has a disadvantage to remove Fe ions from water after wastewater treatment. Thus, iron catalyst was bounded to support such as inorganic and polymer materials. The PVP supporting iron catalyst showed a good catalytic performance in degradation of phenol contained in waste water and iron catalyst supported on ${SO_4}^{2-}$ type PVP (KEX 511) showed the best catalytic performance. Also, reaction kinetic study was carried out in this system. Reaction constants on various catalysts was obtained from the pseudo first order equation. Reaction rate constants with the heterogenized $FeCl_2/PVP$ catalyst is a three-fold smaller than that of $FeCl_2$ catalyst.

The Value of Isotope Nephrography in Carcinoma of Cervix - Follow up Studies of Pre and Post Irradiation (자궁경부암(子宮頸部癌) 방사선치료(放射線治療) 전후(前後) Renogram의 의의(意義))

  • Yoo, H.S.;Suh, J.H.;Park, C.Y.;Choi, B.S.;Jung, S.O.;Kwak, H.M.
    • The Korean Journal of Nuclear Medicine
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    • v.9 no.1
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    • pp.51-58
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    • 1975
  • It is a great value to find an early detection of involvement of ureteric obstruction in the carcinoma of cervix. Little or no knowledge of the condition of the kidneys or the lower urinary tract are able to elucidate by the biochemical studies such as blood nitrogen or urine creatinine in carcinoma of cervix. Findings of urography delineates the condition of urinary tract stasis in the renal pelvis and ureters, however, slight stasis maybe difficult to demonstrate. On the other hand isotope nephrography is accepted as a sensitive method to observe renal function especially in regarding to the excretory function of kidney. It was attempted to analysis the findings of urography conjunction with isotope nephrography in 50 cases of unselected patients with invasive carcinoma of cervix through pre and post irradiation follow up studies. Urography was done as a routine procedure and.analysed emphasising changes of collecting systems and ureter condition. Isotope nephrography was carried out by means of $15{\mu}ci\;I^{131}$-Hippuran injected intravenously and the curves were analysed as follows. Parameter were; time of maximum amplitude ($T_{max}$), half time of maximum amplitude ($T\frac{1}{2}$), Kac and Kex value calculated from these two parameters in Tobe's method. The excretion index by Aurell defines the ratio between the maximum activity and the activity measured on the slope of the third phase ten minites after it has reached its maximum. Results: 1. 28.8% had an abnormal IVP suggestive of ureteric involvement before irradiation therapy and the patient of stage III and IV were the great part. 2. 21.7% had abnormal findings of per-irradiation renogram whom showed normal IVP. The other group showed normal IVP which group also showed normal renogram prior irradiation. 3. The more severe the ureteric involvement, the change of excretion index was greater. 4. Even in stage I and II patient, abnormal renogram was revealed in 12 cases (39.4%) among 31 cases. 5. All cases of TAH showed abnormal findings of IVP and renogram. 6. No. definite change of renogram was obtained just after the irradiation therapy (point $A:8000{\sim}9000rads,\;B:5000{\sim}6000rads,\;Co:11000{\sim}13000rads$). Each 3 month follow up study was performed and comparing with preirradiation study which showed significant changes of excretion index of renogram were 42.8% in $6{\sim}9$ month follow-up and 75% in $9{\sim}12$ month, respectively. 7. It seems to be important to observe the parameter Kex and excretion index of renogram to determine early abnormality of kidney excretory function by means of post-irradiation follow up study.

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Study on the relationship between the potassium activity ratio of paddy soils and potassium uptake by rice plant (답토양(沓土壤)의 가리(加里) Activity ratio와 수도(水稻)의 가리(加里) 흡수(吸收) 특성(特性)에 관(關)한 연구)

  • Kim, Tai-Soon
    • Korean Journal of Soil Science and Fertilizer
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    • v.9 no.4
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    • pp.223-233
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    • 1976
  • The potassium equilibrium activity ratio ($AR^k_e$) and energies of exchange for replacement of ca+Mg by K ($E_k$) were measured for seven paddy soils to investigate their correlations with the exchangeable K (Kex) and uptake of K at different growth stages of rice plant. It was found that $AR^k_e$ had highly significant correlations at 1% level with uptake of K at maximum tillering, heading and harvesting stages, and also with Kex in soils at maximum tillering stage. The larger $AR^k_e$ of soils, the more uptake of K by rice plant. The fact indicates that uptake of K by the plant can be characterized in terms of $AR^k_e$ or energy of exchange of soils. In aspect of energy of exchange, higher uptake of K and yield of grain were observed from -2500 to -3000 calories per chemical equivalent, representing suitable balances between K and Ca+Mg in soils. Low uptake of K was observed at the energies of exchange below -3500 calories per chemical equivalent, which were prevalent in the ordinary acidic soils. From the correlations between energy of exchange and Kex, it can be concluded that at least 0.37 meq. of exchangeable K should be existed in 100g of dried acidic soil to keep suitable balances of K and Ca+Mg. The result shown that application of K adsorbed zeolite to paddy soils increased $AR^k_e$ and consequently brought about higher K uptake and grain yield. Therefore, a reasonable way recommended to get good balance of exchangeable K in the soil is applying 1.7 tonns of K adsorbed zeolite containing 60kg $K_2O$ per hectare.

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Evaluation of the Parameters of Soil Potassium Supplying Power for Predicting Yield Response, K2O Uptake and Optimum K2O Application Levels in Paddy Soils. -II. Determination of Potassium Supplying Power by Gapon equation and Kas/Kai and Response to K2O application (수도(水稻)의 가리시비반응(加里施肥反應)과 시비량추정(施肥量推定)을 위한 가리공급력(加里供給力) 측정방법(測定方法) 평가(評價) -II. Gapon식(式)과 Kas/Kai에 의한 가리공급력(加里供給力) 측정(測定)과 시비반응(施肥反應))

  • Park, Yang-Ho;Ahn, Su-Bong;Park, Chon-Suh
    • Korean Journal of Soil Science and Fertilizer
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    • v.17 no.4
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    • pp.363-370
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    • 1984
  • In order to predict the possible fertilizer requirement from the K supplying capacity of soil, the relative K activity ratio, Kas/Kai and Gapon coefficients, KG. were determined for the soil samples before flooding and at heading stage of rice in pot experiment. These parameters assumed as the K supplying capacity of soils were discussed through correlation with other factors such as grain yields or the amounts of $K_2O$ uptake by the rice plant. The results may be summarized as follows: 1. The KGo values in soils before flooding were 7.8, 6.6, and 7.1, whereas the Kas/Kai values were 1.37, 1.26 and 2.11, respectively, in clay, loam and sandy loam soils. 2. The significant yield responses to the application of potassium fertilizer were observed whenever the KG values in soils at heading stage become larger to the original KG values, regardless of any levels of fertilizer application. 3. The linear correlations between the exchangeable cation ratios [Kex./(Ca+Mg) ex.:me/100g] in soils and the potassium activity ratios ($[K^+]/\sqrt{[Ca^{{+}{+}}+Mg^{{+}{+}}]}$: mole/l) in equilibrium solutions were observed with different linear gradients according to the soil properties. 4. The Kas/Kai in the soils, estimated prior to the experiment, showed high correlations with the grain yields or the amounts of $K_2O$ uptake in the all treatments, while the Kas/Kai and the KGo in the soils at heading stage showed high correlations with the grain yields or the amounts of $K_2O$ uptake in only N 15 Kg/10a treatments. 5. The Kas/Kai and the KGo values determined in the soil at heading stage of rice showed high negative correlation each other and they could be used as soil factors for predicting potassium fertilizer requirement.

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A New Signal Sequence for Recombinant Protein Secretion in Pichia pastoris

  • Govindappa, Nagaraj;Hanumanthappa, Manjunatha;Venkatarangaiah, Krishna;Periyasamy, Sankar;Sreenivas, Suma;Soni, Rajeev;Sastry, Kedarnath
    • Journal of Microbiology and Biotechnology
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    • v.24 no.3
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    • pp.337-345
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    • 2014
  • Pichia pastoris is one of the most widely used expression systems for the secretory expression of recombinant proteins. The secretory expression in P. pastoris usually makes use of the prepro $MAT{\alpha}$ sequence from Saccharomyces cerevisiae, which has a dibasic amino acid cleavage site at the end of the signal sequence. This is efficiently processed by Kex2 protease, resulting in the secretion of high levels of proteins to the medium. However, the proteins that are having the internal accessible dibasic amino acids such as KR and RR in the coding region cannot be expressed using this signal sequence, as the protein will be fragmented. We have identified a new signal sequence of 18 amino acids from a P. pastoris protein that can secrete proteins to the medium efficiently. The PMT1-gene-inactivated P. pastoris strain secretes a ~30 kDa protein into the extracellular medium. We have identified this protein by determining its N-terminal amino acid sequence. The protein secreted has four DDDK concatameric internal repeats. This protein was not secreted in the wild-type P. pastoris under normal culture conditions. We show that the 18-amino-acid signal peptide at the N-terminal of this protein is useful for secretion of heterologous proteins in Pichia.