• The Korean Journal of Physiology and Pharmacology
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• 제2권5호
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• pp.637-644
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• 1998

Previously, we have isolated authentic bombesin and another bombesin like peptide named bombesin like immunoreactivity (BLI)-K2 from the skin of Korean fire-bellied toad, Bombina orientalis. In the present study, we have newly purified three heterogeneous forms of BLI named BLI-K3, BLI-K4, and BLI-K5 from side fractions obtained in previous isolation of bombesin like peptide. The BLIs were separated into five peaks on a column of $C_{18}$ preparative HPLC. Among them, three minor peaks containing BLI-K3, K4, and K5 were purified by means of sequential chromatography on the columns of SP cation exchange HPLC and $C_{18}$ reverse phase HPLC. The purified BLI-K3 and K4 showed high binding affinity to an anti-bombesin serum (LBE 2G-2) with binding potency of 72 and 95%, respectively, relative to that of bombesin. However, they did not possess any distinctive biological activity of bombesin like peptide. On the contrary, the biological activity of BLI-K5 was similar to that of bombesin but its binding affinity to an anti-bombesin serum was low. The results indicate that three heterogeneous forms of BLI were coexpressed with bombesin and BLI-K2 in the skin of B. orientalis. All forms of the purified BLI in the present study were immunologically active but only BLI-K5 possessed the distinctive biological activity of bombesin like peptide.

• Animal cells and systems
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• 제11권2호
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• pp.175-182
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• 2007

The lysozyme II gene of cabbage butterfly Artogeia rapae was cloned from fat body of the larvae injected with E. coli and its nucleotide sequence was determined by the RACE-PCR. It has an open reading frame of 414 bp nucleotides corresponding to 138 amino acids including a signal sequence of 18 amino acids. The estimated molecular weight and the isoelectric point of the lysozyme II without the signal peptide were 13,649.38 Da and 9.11, respectively. The A. rapae lysozyme II (ARL II) showed the highest identity (81%) in the amino acid sequence to Manduca sexta lysozyme among other lepidopteran species. The two catalytic residues ($Glu^{32}$ and $Asp^{50}$) and the eight Cys residue motifs, which are highly conserved among other c-type lysozymes in invertebrates and vertebrates, are also completely conserved. A phylogenetic analysis based on amino acid sequences indicated that the ARL II was more closely related to M. sexta, Hyphantria cunea, Heliothis virescens, and Trichoplusia ni lysozymes. The ARL II gene was expressed in Spodoptera frugiperda 21 insect cells and the recombinant ARL II (rARL II) was purified from cell-conditioned media by cation exchange column chromatography and reverse phase FPLC. The purified rARL II was able to form a clear zone in lysoplate assay against Micrococcus luteus. The lytic activity was estimated to be 511.41 U/mg, 1.53 times higher than that of the chicken lysozyme. The optimum temperature for the lytic activity of the rARL II was $50^{\circ}C$, the temperature dependency of the absolute lytic activity of rARL II was higher than that of the chicken lysozyme at low temperatures under $65^{\circ}C$.

• 한국식품과학회지
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• 제31권4호
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• pp.1120-1123
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• 1999

본 연구는 멸치 자건품의 선상 가공시 유출되는 자숙액 중에 다량으로 함유되어 있는 타우린을 분리, 정제하고자 하였다. 원료 자숙액 중의 타우린 함량은 112.5 mg/100g으로서 총유리아미노산 함량의 38.8%를 차지하였다. 이 자숙액을 Amberlite IR-120과 Amberlite IR-400 column에 순차적으로 적용하여 수율이 79.2%, 순도가 89.9%인 타우린을 얻었다. 또한 이를 다시 에탄올로 정제한 결과, 수율이 29.5%, 순도가 98.1%인 고순도 타우린 결정을 얻을 수 있었다.

• Asian Pacific Journal of Cancer Prevention
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• 제14권7호
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• pp.4161-4165
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• 2013

Objective: To investigate discriminating protein patterns and potential biomarkers in serum samples between pre/postoperative pancreatic cancer patients and healthy controls. Methods: 23 serum samples from PC patients (12 preoperative and 11 postoperative) and 76 from healthy controls were analyzed using matrix-assisted laser desorption and ionization time-of-flight mass spectrometry (MALDI-TOF MS) technique combined with magnetic beads-based weak cation-exchange chromatography (MB-WCX). ClinProTools software selected several markers that made a distinction between pancreatic cancer patients and healthy controls. Results: 49 m/z distinctive peaks were found among the three groups, of which 33 significant peaks with a P < 0.001 were detected. Two proteins could distinguish the preoperative pancreatic cancer patients from the healthy controls. About 15 proteins may be potential biomarkers in assessment of pancreatic cancer resection. Conclusion: MB-MALDI-TOF-MS method could generate serum peptidome profiles of pancreatic cancer and provide a new approach to identify potential biomarkers for diagnosis and prognosis of this malignancy.

• 한국수산과학회지
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• 제44권5호
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• pp.478-483
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• 2011

An antimicrobial material was purified from the acidified whole body extract of the jellyfish Nemopilema nomurai by using C18 reversed phase and cation-exchange high performance liquid chromatography (HPLC). Whole body extract and the purified compound (JAP-1) showed potent antimicrobial activities against a wide range of microorganisms including Escherichia coli D31, Bacillus subtilis, Streptococcus iniae and Candida albicans, without significant hemolytic activity. Treatment of JAP-1 with trypsin completely abolished all antibacterial activity against Bacillus subtilis, suggesting that JAP-1 is likely to be a proteinaceous antibiotic. The molecular weight of JAP-1 was determined to be 680.10 Da by MALDI-TOF mass spectroscopy.

• Biotechnology and Bioprocess Engineering:BBE
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• 제11권5호
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• pp.442-448
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• 2006

Human lactoferrin (hLf) is an iron-binding glycoprotein that has been considered to play many biological roles in the human, including the stimulation of the immune system, antimicrobial and anti-inflammatory effects, and regulation of iron absorption. We generated transgenic Siberian ginseng (Acanthopanax senticosus) cell cultures producing a functional hLf protein using the signal peptide sequence from the endoplasmic reticulum and driven by an oxidative stress-inducible SWPA2 promoter which is highly expressed in plant cell cultures. The production of hLf increased proportionally to cell growth and showed a maximal level (up to 3.6% of total soluble protein) at the stationary phase in suspension cultures. Full-length hLf protein was identified by immunoblot analysis in transgenic cell cultures of Siberian ginseng. Recombinant hLf (rhLf) was purified from suspension cells of Siberian ginseng by ammonium sulfate precipitation, cation-exchange and gel filtration chromatography. N-terminal sequences of rhLf were identical to native hLf (nhLf). The overall monosaccharide composition of rhLf showed the presence of plant specific xylose while sialic acid is absent. Antibacterial activity of purified rhLf was higher than that of nhLf. Taken together, we anticipate that medicinal Siberian ginseng cultured cells, as demonstrated by this study, will be a biotechnologically useful source for commercial production of functional hLf not requiring further purification.

• 분석과학
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• 제13권6호
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• pp.781-788
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• 2000

수소화물 생성-유도결합플라스마 원자방출분광법(HG-ICP-AES)을 이용하여 모의사용후 핵연료(SIMFUEL) 중의 텔루르를 정량하였다. 염산과 $NaBH_4$의 농도 및 주입속도와 같은 변수들을 최적과 한 다음 각각 우라늄, 팔라듐, 루테늄, 로듐 및 몰리브덴의 간섭 정도를 조사하였다. 이들 원소, 특히 팔라듐으로부터 간섭을 줄이기 위하여 가리움제로 thiourea를 사용하였다. 모의사용후 핵연료로부터 텔루르를 양이온 교환 크로마토그래피로 분리한 다음 각각 HG-ICP-AES와 유도결합플라스마 질량분석법(ICP-MS)으로 측정하였다. 우라늄 매트릭스로부터 텔루르를 분리하지 않고 바로 전자로 측정한 결과와 분리한 다음 측정한 값의 상대편차는 ICP-MS의 결과를 기준으로 5.6%와 -1.2%이었다.

• Archives of Pharmacal Research
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• 제19권2호
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• pp.153-159
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• 1996

A new streptothricin family antibiotic producing Streptomyces spp. SNUS 8810-111 was isolated from a soil sample. Study of its morphological and physiological characters indicated that the antibiotic producing organism was a Streptomyces spp. Taxonomical studies suggested that the organism might belong to the genus streptomyces gougeroti. The organism produced antibiotics most in calcium carbonate-tryptic soy broth. The active principles were recovered from the broth with a cation exchange resin and eluted from the resin with HCI. Cellulose column chromatography gave two active principles.$^1H-^1H$ Homo-COSY study on the first compound revealed four structural components. Total hydrolysis of the antibiotic with HCI allowed isolation of $\beta-lysine$. From these data the antibiotic was found to be streptothricin D. The other compound showed one additional signal in the .$^1H$NMR and the $^{13}C$ NMR spectra. The signal was from a methyl group attached to a nitrogen atom. Comparison of the NMR signals with those of streptothricin D suggested that the compound was N-methyl-streptothricin D which was a new compound in the family of streptothricin antibiotics.

• 한국식품과학회지
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• 제35권2호
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• pp.260-265
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• 2003

본 연구에서는 수율과 protease 제거정도 그리고 분리정제 비용을 고려하여 암모늄 설페이트 침전과 Monoplus S100 수지로 처리한 효소를 밀가루에 적용하여 반죽 형성 시 물리적 특성개선 효과를 조사하였다. 암모늄설페이트 침전, 이온 교환수지, 겔 크로마토그래피를 통한 정제된 효소의 분자량은 SDS-PAGE상에서 38,000으로 판정되었고 단일밴드로 나타나 정제도가 높음을 알 수 있었다. Farinograph를 이용하여 측정한 밀가루 반죽의 특성변화에서 저항성의 지표가 되는 안정도는 대조구가 4분, 조효소는 대조구보다 짧은 3.5분, 정제된 효소는 대조구보다 길게 측정되었다. 반죽의 약화도는 조효소의 경우 대조구보다 높게 측정되었고, 정제된 효소의 경우 대조구보다 낮은 값을 나타냈다. 이 결과로부터 조효소의 경우 protease 존재로 인하여 반죽의 분해가 진행됨을 알 수 있었고, 정제된 효소 첨가로 인하여 물성이 향상됨을 알 수 있었다. 대조군의 밀가루 반죽의 valorimeter 값은 52로 측정되었고, 정제된 효소첨가에 의해 증가되었다. S. mobaraensis가 분비하는 transglutaminase의 정제단계를 조절하고 최적화 시킬 경우 국내산 밀가루의 반죽특성의 효과를 기대할 수 있다고 판단된다.

• 한국미생물·생명공학회지
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• 제42권1호
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• pp.25-31
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• 2014

본 연구에서는 바이오매스 유래 생리활성물질인 (+)-dihydromyricetin을 효율적으로 정제하기 위하여, 반응액 부피당 표면적(S/V)이 증가된 새로운 개념의 분별침전공정을 도입하였다. 분별침전 24시간에서 반응기 내부 표면적을 증가시키지 않은 경우 순도와 수율은 각각 80.0%와 70.0%인 반면 표면적 증가를 위해 양이온교환수지인 Amberlite 200을 첨가한 경우 순도와 수율은 각각 90.2%와 90.9%로 가장 효과적인 표면적증가물질임을 알 수 있었다. 특히 Amberlite 200의 경우 상대적으로 짧은 침전시간(16시간)에 높은 수율(>90%)로 (+)-dihydromyricetin을 얻을 수 있어 침전에 소요되는 시간을 효과적으로 단축시킬 수 있었다. 동일한 침전시간에서는 표면적증가물질을 첨가하지 않은 경우에 비해 표면적증가물질을 첨가한 경우 침전물의 입자 크기가 감소함을 알 수 있었다. 이러한 연구결과는 기존의 분별침전 방법에 의한 (+)-dihydromyricetin 정제공정의 문제점을 개선함으로써(+)-dihydromyricetin의 대량생산에 매우 유용하게 활용될 수 있을 것으로 판단된다.