중금속 제거 및 pH 상승을 위한 폐달걀껍질의 재활용

조영수,최영락,손희정,김은호 東亞大學校生命資源科學大學附設 農業資源硏究所 1997 農業生命資援硏究 Vol.6 No.1

In the present, batch test was conducted to evaluate the neutralization and adsorption of heavy metals from wastewater using waste egg shells. Neutralization and removal rate of heavy metals were excellent in the increase of waste concrete amounts and a small size. It seemed that adsorption efficiencies of heavy metals were influenced by solubility. As a result on the experiments of Freundlich isotherm, the adsorption capacities(k) were Cr 3.11, Cu 2.61, Mn 3.02 and Pb 0.95, respectively and the adsorption intensities(1/n) were Cr 0.35, Cu 0.44, Mn 0.4 and Pb 0.41, respectively. In view of these results, it showed that wastes containing the similar compositions as waste egg shells could utilize the neutralization and adsorption of heavy metals in wastewater.

Candida tropicalis로 부터 분리된 자율복제기점을 이용한 Candida maltosa와 Saccharomyces cervisiae에서의 shuttle-vector의 구축

최용락,조영수,차재영 東亞大學校附設遺傳工學硏究所 1994 遺傳工學硏究 Vol.- No.1

무포자 효모인 Candida속의 효모는 aikane계 화합물을 이용하며 cellubiose나 xylose를 동화하는 등 산업적으로 이용가능성이 높다. Candida tropicalis는 세포질 인자인 plasmid가 발견되지 않으므로 염색체 유래의 자율복제기점을 non-replicative plasmid인 YIp5에 삽입시켜 S. cevisiae YNN 27에서 분리하였다. 분리된 자율 복제 기점을 Hind III과 EcoR I제한 효소로 절단하고 각각 2.7kb와 2.3kb fragment를 회수한 후 동일 제한 효소로 절단한 YIp5와 YIp32에 ligation시켜 pIKS523, pIKS527및 pIKC27을 구축하였다. 특히, pIKC27은 2.7kb의 자율복제 기점을 가진 plasmid로서 S. cerevisiae DBY746과 C.maltosa J288에서 높은 형질 전환체를 나타내었다. 기존의 plasmid와 안정성을 비교하였으며, 여러 제한효소에 의해서 얻어진 결과로서 상세한 restrition map을 작성하였다. Candida species are special interest because of its ability to metabolize hydrocarbon, covet this material into single cell protein and assimilate cellobiose and xylose as substrate. Cadida tropicalis has no native plasmid similar to 2㎛ plasmid. Therefore, Candida tropicalis is regarded as a new yeast host. An 8-kb fragment was isolated from Sal I digest of Candida tropicalis IFO 0518 genomic DNA which conferred the property of autonomous replication in Saccharomyces cerevisiae YNN 27. The vectors for gene manipulation of Yeast, pIKS 523, pIKS 527 and pIKC 27, were constructed by combinding the ARS fragment and integration plasmid YIP 5 and YIP 32, respectively. One of the recombinant plasmids, pIKC 27 (9.4kb), was capable of autonomous replication in both S. cerevisiae DBY 746 to Leu+ at a frequency of 600 transformants per ㎍ DNA, and transformed Candida maltosa J288 to Leu+ at a frequency of 230 transformants per ㎍ DNA. These results indicate that the 2.7 kb ARS element was necessary for high frequency transformation and autonomous plasmid replication in both S. Cerevisiae and C. maltosa.

CRP* 의존성 maltose 대사 촉진 유전자 sfs4의 클로닝 및 염기배열 결정

최용락,정희태,조무제,정수열 東亞大學校附設遺傳工學硏究所 1996 遺傳工學硏究 Vol.- No.3

CRP (cAMP receptor protein)은 cAMP와 결합하여 cAMP-CRP 복합체를 형성하여 전사조절의 조절인자로서 작용한다. crp 유전자에 변이를 도입하여 cAMP의 비존재 상태에서 cAMP-CRP와 비슷한 기능을 가진 crp* 유전자가 도입된 대장균 MK2001 (crp*¹, cya::km)을 숙주로 사용하여 cAMP 혹은 cGMP의 비존재하에서도 mal 유전자의 발현을 촉진시키는 유전자 sfs (sugar fermentation stimulation) 수 종을 클로닝하였다. 본 실험에서는 이미 밝혀진 nlp (Ner like protein) 유전자와 같이, sfs의 새로운 유전자를 탐색하여, 그 중 sfs4의 2126bp 전 염기배열을 결정하고, 잠정적인 sfs4의 promoter 영역에는 CRP 단백질과의 결합 DNA 공통 염기배열(5' AAT TGTGA ACACCA TCACC CGT 3')이 존재함을 확인했다. lacZ 융합 유전자를 작성하여 TP2010R1과 MK2001의 균주에서 cAMP를 첨가할 경우 각각 2.3배, 1.8배의 β-galactosidase 활성이 증가 하는 것으로 보아 sfs4는 cAMP-CRP에 의해 발현 조절을 받는 것으로 나타났다 In Escherichia coli, CRP forms a complex with cAMP and acts as a transcriptional regulator of many genes, including sugar metabolism operons. The E. coli MK2001, which is introduced the altered crp*¹, is functional in the expression of lac, ara and man, in the absence of cAMP. However, the expression of mal gene is fully activated by the addition of cAMP or cGMP. The object of the study is cloning of the sfs (sugar fermentation stimulation) genes, which was involved in regulation of mal gene expression with the altered crp*¹gene, and structural analysis and characterization of the genes at the molecular level. We have cloned 5 different E. coli genes which stimulate the maltose metabolism in a crp*¹, cya::km (MK2001) background. Newly identified genes were designated as sfs. One of the sfs genes (pPC1), located at the 53.2 min map position in the E. coli chromosome, was further analyzed. Expression of the genes, which is involved in maltose metabolism, malQ (amylomaltase), was increased to 5.8-fold in the presence of a plasmid, pAP5, containing the subcloned sfs4 gene. The nucleotide sequence of a common 2,126bp segment of the pPCM1 was determined and two open reading frames (ORF1 and ORF2) were detected. The ORF1 encodes the sfs4 gene and ORF2 encodes a truncated protein. Potential CRP binding site is located in the upstream of the putative promoter in the regulatory region. Expression of the cloned sfs4 gene was positively regulated by the cAMP-CRP complex.

Compared chitinase deleted binding domain with chitinase from Bacillus atrophaeus TBM1652-1 in E.coli

Yong-Seok Lee,Ju-Soon Yoo,Soo-Yeol Chung,Young-Choon Lee,Young-Su Cho,Yong-Lark Choi 한국생명과학회 2005 한국생명과학회 심포지움 Vol.45 No.-

Cloning, purification and characterization of chitosanase from Bacillus sp. DAU101

Yong-Seok Lee,Ju-Soon Yoo,Soo-Yeol Chung,Young-Choon Lee,Young-Su Cho,Yong-Lark Choi 한국생명과학회 2006 한국생명과학회 심포지움 Vol.46 No.-

cDNA Cloning of Alfalfa (Medicago Sativa L. Vernal) Leghemoglobin Structural Genes

CHO, MOO JE,YUN, HAN DAE,CHOI, YONG LARK,CHOI, YOUNG JU,LEE, KYUNG SANG,BRILL, WINSTON J. 경상대학교 유전공학연구소 1984 遺傳工學硏究所報 Vol.3 No.-

Complementary DNA were synthesized by the leghemoglobin message rich 9S, 18S, and 28S-mRNA fractions prepared from the poly(A)^+-mRNA of alfalfa root nodules. The synthetic SalI linkered double stranded cDNA was inserted into the SalI site of the pBR322, and transformed in E. coil HB101. The leghemoglobin cDNA clones were screened by colony hybridization and Southern blot hybridization with the soybean leghemoglobin cDNA as a probe, and further screened by hybrid-released and hybrid-arrested translation. One alfalfa leghemoglobin cDNA clone was selected., and restriction map and fingerprints of the cDNA from the selected clone were analyzed.

Purification of Leghemoglobin mRNA from Alfalfa (Medicago Stiva L, Vernal) Root Nodules and In Vitro Translation

Cho, Moo Je,Yang, Min Suk,Yun, Han Dae,Choi, Yong Lark,Choi, Young Ju,Lee, Kyung Sang 경상대학교 유전공학연구소 1984 遺傳工學硏究所報 Vol.3 No.-

The five components of leghemoglobin from alfalfa root nodules were purified by DEAE-cellulose (DE52) and Sephadex G-100 chromatography and antiserum against the purified alfalfa leghemoglobin was prepared. RNA was extracted from alfalfa root nodules and poly(A)^+mRNA was purified by oligo(dT)-cellulose affinity chromatography. The leghemoglobin messages were observed to be 23% of the poly(A)^+mRNA. The poly(A)^+mRNA was fractionated by sucrose density gradient centrifugation and the mRNA had 9, 18 and 28S value populations with 58, 21 and 36%of the leghemoglobin messages.

Characterization of the Small Cryptic Plasmid , pGD2 , of Klebsiellia sp. KCL - 2

Choi, Yong Lark,Chung, Soo Yeol,Lee, Young Choon,Yoo, Ju Soon,Kim, Hae Sun,Cho, Young Soo 생화학분자생물학회 1998 BMB Reports Vol.34 No.6

One of the cryptic plasmids from the oil degrading bacterium Klebsiella sp. KCL-2, the small plasmid pGD2, has been identified and characterized. This plasmid has a size of 3.6 kb with unknown functions. We constructed the recombinant plasmid pMGD2. The nucleotide sequences of the plasmid were determined and two open reading frames were detected. ORF1 encodes a replication initiator protein (RepA), which has a high degree of homology with the protein of ColE2 plasmid. The product encoded by ORF2 showed a high similarity with the transposase protein of IS5. IS5 is 1195 by long and contains an inverted terminal repetition of 16 bp with one mismatch. Stem-loop structures in the 5'untranslated region of the repA suggest that a putative gene, incA, is located in a complementary strand to the leader region of the repA mRNA.

AMP-activated protein kinase negatively regulates Fc□RI-mediated mast cell signaling and anaphylaxis in mice

( Seung Lark Hwang ),( Xian Li ),( Yue Lu ),( Ye Jin ),( Yong Tae Jeong ),( Yong Deuk Kim ),( In Kyu Lee ),( Yoshitaka Taketomi ),( Hiroyasu Sato ),( You Sook Cho ),( Makoto Murakami ),( Hyeun Wook Ch 영남대학교 약품개발연구소 2014 영남대학교 약품개발연구소 연구업적집 Vol.24 No.0

Background: Aggregation of FcεRI activates a cascade of signaling events leading to mast cell activation, followed by inhibitory signals that turn off the activating signals. However, the overall view of negative signals in mast cells is still incomplete. Although AMP-activated protein kinase (AMPK), which is generally known as a regulator of energy metabolism, is also associated with anti-inflammation, little is known about the role of AMPK in mast cells. Objectives: We investigated the role of AMPK and its regulatory mechanism in mast cells. Method: The roles of AMPK in FcεRI-dependent activation of bone marrow-derived mast cells (BMMCs) were evaluated by using chemical agents, small interfering RNAs (siRNAs), or adenovirus that modulated the activity or expression of AMPK signaling components. In addition, AMPKα2(-/-) mice were used to verify the role of AMPK in anaphylactic models. Results: FcεRI signaling and associated effector functions in BMMCs were suppressed by the AMPK activator 5-aminoimidazole-4-carboxamide-1-β-4-ribofuranoside (AICAR) and were conversely augmented by siRNA knockdown of AMPKα2 or liver kinase B1 (LKB1), an upstream kinase of AMPK. Furthermore, AMPKα2 deficiency led to increased FcεRI-mediated BMMC activation and anaphylaxis that were insensitive to AICAR, whereas enforced expression of AMPKα2 in AMPKα2(-/-) BMMCs reversed the hypersensitive FcεRI signaling to normal levels. Pharmacologic inhibition or siRNA knockdown of Fyn mimicked AMPK activation, suggesting that Fyn counterregulates the LKB1-AMPK axis. Mechanistically, Fyn controlled AMPK activity by regulating LKB1 localization. Conclusions: The Fyn-regulated LKB1-AMPK axis acts as a novel inhibitory module for mast cell activation, which points to AMPK activators as therapeutic drugs for allergic diseases. (J Allergy Chin Immunol 2013;132: 729-36)

Medicinal Chemistry : ERK1/2 antagonize AMPK-dependent regulation of FcεRI-mediated mast cell activation and anaphylaxis

( Seung Lark Hwang ),( Yue Lu ),( Xian Li Msc ),( Yong Deuk Kim ),( You Sook Cho ),( Yurn Dong Jahng ),( Jong Keun Son ),( Youn Ju Lee ),( Won Ku Kang ),( Yoshitaka Taketomi ),( Makoto Murakami ),( Ta 영남대학교 약품개발연구소 2015 영남대학교 약품개발연구소 연구업적집 Vol.25 No.-

Background: Extracellular signal-regulated kinases 1/2 (ERK1/2) make important contributions to allergic responses via their regulation of degranulation, eicosanoid production, and cytokine expression by mast cells, yet the mechanisms underlying their positive effects on FcεRI-dependent signaling are not fully understood. Recently, we reported that mast cell activation and anaphylaxis are negatively regulated by AMP-activated protein kinase (AMPK). However, little is known about the relationship between ERK1/2-mediated positive and the AMPK-mediated negative regulation of FcεRI signaling in mast cells. Objective: We investigated possible interactions between ERK1/2 and AMPK in the modulation of mast cell signaling and anaphylaxis. Methods: Wild-type or AMPKa22/2 mice, or bone marrow. derived mast cells obtained from these mice, were treated with either chemical agents or small interfering RNAs that modulated the activity or expression of ERK1/2 or AMPK to evaluate the functional interplay between ERK1/2 and AMPK in FcεRI-dependent signaling. Results: The ERK1/2 pathway inhibitor U0126 and the AMPK activator 5-aminoimidazole-4-carboxamide-1-b-4-ribofuranoside similarly inhibited FcεRI-mediated mast cell signals in vitro and anaphylaxis in vivo. ERK1/2-specific small interfering RNA also mimicked this effect on FcεRI signals. Moreover, AMPKa2 knockdown or deficiency led to increased FcεRI-mediated mast cell activation and anaphylaxis that were insensitive to U0126 or activator 5-aminoimidazole-4-carboxamide-1-b-4-ribofuranoside, suggesting that the suppression of FcεRI signals by the inhibition of the ERK1/2 pathway relies largely on AMPK activation. ERK1/2 controlled AMPK activity by regulating its subcellular translocation. Conclusions: ERK1/2 ablated the AMPK-dependent negative regulatory axis, thereby activating FcεRI signals in mast cells. (J Allergy Clin Immunol 2014;134:714-21.)