• Title/Summary/Keyword: enzymatic technique

Search Result 73, Processing Time 0.029 seconds

Overexpression and Characterization of appA Phytase Expressed by Recombinant Baculovirus-Infected Silkworm

  • CHEN YIN;ZHU ZHONGZE;LIN XU'AI;YI YONGZHU;ZHANG ZHIFANG;SHEN GUIFANG
    • Journal of Microbiology and Biotechnology
    • /
    • 제15권3호
    • /
    • pp.466-471
    • /
    • 2005
  • An Escherichia coli strain with high phytase activity was screened from pig excreta. The phytase gene, appA, was amplified by PCR technique. To obtain large amounts of appA phytase, the appA gene was subcloned into the baculovirus transfer vector pVL1393 under the control of the Polyhedrin promoter. The recombinant baculovirus harboring the appA gene was obtained after co-transfection and screening. The early $5^{th}$ instar larvae of silkworm were infected with the recombinant virus. Using this system, the appA phytase was overproduced up to 7,710 U per ml hemolymph. SDS-PAGE analysis revealed the baculovirus-derived appA phytase to be approximately 47 kDa in size. The optimal temperature and pH of the expressed phytase were $60^{\circ}C$ and pH 4.5, respectively. The enzymatic activity was increased by the presence of 1 mM $Ca^{2+}$, 1 mM $Mn^{2+}$, or $0.02\%$ Triton X-100.

밀기울배지를 이용한 Bacillus macerans의 Cyclodextrin Glucanotransferase 생산과 효소특성 (Purification and Enzymatic Properties of Cyclodextrin Glucanotransferase from Bacillus macerans Cultivated in Wheat-bran Medium)

  • 선우양일;안태진
    • KSBB Journal
    • /
    • 제9권5호
    • /
    • pp.499-505
    • /
    • 1994
  • Bacillus macerans cyclodextrin glucanotrans­f ferase(CGTase)를 전분흡착법과 DEAE--cellulose 칼럼 크로마토그래피로 정제하였다. 효소의 분자량 은 67,000이였고 monomer였다. 정제된 효소는 전 분을${\alpha}$-, ${\beta}$-, ${\gamma}$-CD 로 전환시켰으며, CD생성비율은 각각 1 : 1.68: 0.32였다. ${\alpha}$-CD와 D-glucose의 coupling반응 초기에는 maltohexose가 주로 생성되었고, 그 후에 다른 oligosaccharide들이 생성되었다 .. a­C CD의 가수분해반응 초기에는 주로 maltotetrose가 생성되였고 그 이후에는 소량의 다른 이Igosa­C ccharide들이 생성되었다. 정제된 효소의 좋은 기질인 maltotriose 로부터 maltosyl 이나 D-glucopy ranosyl group이 전이될 수 있는데, 본 연구에서는 D정lucosyl transfer가 우세하였다.

  • PDF

Regulatory Mechanism of Lysine Biosynthetic Genes in Escherichia coli

  • Joe, Min-Ho;Mun, Hyo-Young;Hong, Mi-Ju;Kim, Seong-Jun;Park, Young-Hoon;Rhee, Sang-Ki;Kwon, Oh-Suk
    • 한국미생물생명공학회:학술대회논문집
    • /
    • 한국미생물생명공학회 2004년도 Annual Meeting BioExibition International Symposium
    • /
    • pp.236-241
    • /
    • 2004
  • In Escherichia coli, L-lysine biosynthetic pathway is composed of nine enzymatic reactions. It has been well established that most of the lysine biosynthetic genes are regulated by the lysine availability, even though they are all scattered around the chromosome without forming any multigenic operon structure. However, no transcriptional regulatory mechanism has been identified except for the activation of lysA gene by the LysR. In this study, changes in transcriptome profiles of wild type cells and lysR deletion mutant cells grown in the absence or presence of lysine were investigated by using DNA microarray technique. Microarray data analysis revealed three groups of genes whose expression varies depending on the availability of lysine or LysR or both. To further examine the regulatory patterns of lysine biosynthetic genes, lacZ operon fusions were constructed and their expression was measured under various conditions. Obtained results strongly suggest that there is an additional regulatory mechanism which senses the lysine availability and coordinates gene expression.

  • PDF

Design and Implementation of Magnetic Stimulation Device Suitable for Herpes Zoster and Post Herpetic Neuralgia

  • Tack, Han-Ho;Kim, Gye-Sook;Kim, Whi-Young
    • 한국정보기술학회 영문논문지
    • /
    • 제10권2호
    • /
    • pp.199-214
    • /
    • 2020
  • An important technique of the present invention is primarily to parallel light detection, self-pulse therapy after diagnosis. Herpes zoster is a disease caused by varicella zoster virus, and the virus that has been latent in the dorsal root ganglion that controls the skin segment loses its immune system and physically damages it. It is an acute skin disease in which acute pain and bullous rash occur along the sensory ganglia, which are rehab by inducers such as malignant tumors. Dorsal root ganglion after complete recovery of varicella, relapsed after incubation in brain ganglion, latent virus sometimes suppressed activity by cell mediated immunity, and in cell ganglion with reduced cellular immunity. It proliferates and destroys neurons, causing pain while forming a rash and blisters. This can reduce cell necrosis and increase the phagocytosis and enzymatic activity through the movement of ions through the cell membrane, depolarization and membrane potential change, growth factor secretion, calcium ion transfer, chondrocyte synthesis, etc., And may offer treatment options for lesions of herpes zoster and post-herpetic neuralgia (PHN).Therefore, according to the present research, the diagnosis and treatment device of treating paing for herpes zoster and post-herpetic pain can be implemented in the early stage of herpes zoster, and conventional analgesic regulation, anti-inflammatory effect, post-herpetic neuralgia.

백미의 총 식이섬유함량 예측 모델 개발을 위한 퓨리에변환 근적외선분광계의 적용 (Application of Fourier Transform Near-Infrared Spectroscopy for Prediction Model Development of Total Dietary Fiber Content in Milled Rice)

  • 이진철;윤연희;은종방
    • 한국식품저장유통학회지
    • /
    • 제12권6호
    • /
    • pp.608-612
    • /
    • 2005
  • 친환경적이면서 신속한 비파괴 분석방법인 FT-NIR를 이용하여 백미의 총식이섬유(TDF)함량 예측모델을 개발하였다. 백미는 국내산으로 전남지방에서 재배된 47개 품종과, 시중 유통 중인 13개 브랜드 미에 대해서 AOAC 방법에 준한 효소법에 의해 TDF 함량을 분석하였다. 습식 분석된 TDF함량의 범위는 $1.17-1.92\%$ 이었다. FT-NIR로 측정된 스펙트럼의 검량식은 빛의 산란 효과를 최소화하기 위해 수학적 처리를 하였고, 몇 개의 특정 파장이 아닌 전 파장 영역(1,000-2,500 nm)에 대해서 PLS법으로 작성하였다. 얻어진 검량식의 정확도는 상관계수(r), SEE 및 SEP로 확인하였다. 백미 중 총 식이섬유 함량에 대한 회귀분석을 행한 결과, 검량식의 r은 0.9705, SEE는 0.0464, 검증식의 bias는 -0.0006, SEP가0.0604로 측정 정확도가 우수하여 실제 적용이 가능함을 보여주었다.

각질세포와 섬유모세포가 배양된 고분자 미립구를 이용한 피부재생

  • 곽소정;김상수;전현욱;이영무;박문향;송강원;최차용;김병수
    • 한국생물공학회:학술대회논문집
    • /
    • 한국생물공학회 2003년도 생물공학의 동향(XII)
    • /
    • pp.92-94
    • /
    • 2003
  • 각질세포의 배양법이 개발 후 피부 결손 부위의 치료에 인체 각질세포를 배양하여 얻어진 표피를 이식하는 방법부터 세포부유물을 도포하는 방법들은 Trypsin 처리 과정을 거치면서배양된 세포의 부착능력을 가진 단백질이 손상되어 성공적인 피부 재생이 불가능하다. 본 실험에서는 이러한 효소처리의 단점을 보완하기 위해서 고분자 미립구에 세포를 효소처리 과정없이 동물실험을 한 결과 21일 후에는 대조군에 비해서 표피가 완벽하게 재생되었고 이식 후 세포의 부착면적을 늘리고 생착율을 높일 수 있었다. 이 연구에서 효소처리 없이 고분자 미립구에서 배양된 세포를 이용한 인공피부 재생이 효과적인 것을 보여주었다. 이러한 모델은 앞으로 화상이나 궤양으로 인한 피부 손실 부위 치료에 사용될 수 있을 것으로 생각된다.

  • PDF

Skin regeneration by spray of fibrin gel containing keratinocytes and epidermal growth factor

  • Gwak, So-Jung;Kim, Sang-Soo;Choi, Cha-Yong;Kim, Byung-Soo
    • 한국생물공학회:학술대회논문집
    • /
    • 한국생물공학회 2003년도 생물공학의 동향(XIII)
    • /
    • pp.356-358
    • /
    • 2003
  • 각질형성세포의 배양법이 개발 후 피부 결손 부위의 치료에 인체 각질형성세포를 배양하여 얻어진 표피를 이식하거나 세포부유물을 도포하는 기존의 방법들은 trypsin 처리 과정을 거치면서 배양된 세포의 부착능력을 가진 단백질이 손상되어 성공적인 피부 재생이 불가능하다. 이 연구에서는 면역결핍 생쥐 모델에서 인간 피부 각질형성에포와 EGF-피브린 고분자 혼합물을 분사하여 피부의 전층 상처를 재생하였다. 조직학 검사와 면역화학검사를 통하여 각질형성세포와 EGF-피브린 고분자를 함께 분사한 경우 이식된 인간 표피세포에 의한 빠른 표피재생을 확인할 수 있었다. 이 피부 재생술은 기존의 배양된 인공피부 sheet의 이식을 이용한 피부재생법에 비해 여러 가지 장점을 가지고 있으므로, 앞으로 화상이나 피부궤양과 같은 피부결손의 효율적인 새로운 치료법으로 사용되어질 수 있을 것이다.

  • PDF

Presence of Transgenic Genes and Proteins in Commercial Soybean Foods from Mexican Grocery Stores

  • Cruz-Flores, Yendi Arely;Rodriguez-Herrera, Raul;Aguilar-Gonzalez, Cristobal Noe;Contreras-Esquivel, Juan Carlos;Reyes-Vega, Maria de la Luz
    • Food Science and Biotechnology
    • /
    • 제17권5호
    • /
    • pp.1092-1096
    • /
    • 2008
  • Commercial food products from major cities of Coahuila, Mexico were screened to identify residues of transgenic deoxyribonucleic acid (DNA) and/or proteins. After performed, an inventory on all products that contained a soybean-based ingredient in a commercial grocery store in the city of Saltillo, Coahuila, Mexico, 245 food products were identified and grouped in 15 classes according to the soybean ingredient as well as the manufacturing process used for their elaboration. Similar sampling was made for the different food classes in the cities of Monclova, Piedras Negras, and Torreon. A total of 88 samples were analyzed and DNA was extracted by the hexadecyltrimethyl-ammonium bromide (CTAB) technique with slight modification to obtain better DNA quality (1). In addition, segments of the transgenic genes one that codifies for 5-enolpyruvylshikimate-3-phosphate synthase (epsps), cry 1A, and the cauliflower mosaic virus (CaMV) promoter were amplified using polymerase chain reaction (PCR). The transgenic proteins 5-enolpyruvylshikimate-3-phosphate synthase (CP4 EPSPS) and insecticidal crystal protein (Cry 1Ab/Ac) were identified using double antibody sandwich-enzymatic linked immunoassay analysis (DAS-ELISA). Presence of transgenic genes and/or proteins was identified in 35.3% of the commercial products samples.

Effects of Phenylephrine on the Excitability of Medial Vestibular Nuclear Neurons in Rats

  • Jeong, Han-Seong;Huh, Hae-Ryong;Jang, Myung-Joo;Hong, Seol-Hee;Jang, Su-Jeong;Park, Jin;Lee, Seung-Han;Kim, Jae-Ha;Park, Jong-Seong
    • The Korean Journal of Physiology and Pharmacology
    • /
    • 제10권3호
    • /
    • pp.131-135
    • /
    • 2006
  • Coeruleo-vestibular pathway which connects locus coeruleus and vestibular nuclei is noradrenergic. This study was designed to elucidate the effects of phenylephrine on the spontaneous activity of acutely isolated medial vestibular nuclear neurons of rat by whole-cell patch-clamp technique. Sprague-Dawley rats, aged 14 to 16 days, were used. After enzymatic digestion, dissociated medial vestibular neurons were transferred to a recording chamber mounted on an inverted microscope, and spontaneous action potentials were recorded by standard patch-clamp techniques. In current-clamp mode, the frequency of spontaneous action potential of medial vestibular nuclear neurons was decreased by phenylephrine (n=15). Phenylephrine increased the amplitude of afterhyperpolarization without changes in the resting membrane potential and spike width. In voltage-clamp mode, the whole potassium currents of the medial vestibular nuclear neurons were increased by phenylephrine (n=12). These experimental results suggest that ${\alpha}-receptor$ mediates the inhibitory effects on the neuronal activity of the medial vestibular nuclear neuron.

발현성 고혈압쥐의 관상순환 기능과 심장근의 에너지 대사에 관한 생체외 에너지원의 관류 연구 (Substrate-Perfusion Studies on Coronary Circulation and Myocardial Energy Metabolism in Spontaneously Hypertensive Rat Hearts)

  • 김은지
    • Journal of Nutrition and Health
    • /
    • 제28권2호
    • /
    • pp.115-126
    • /
    • 1995
  • The effects of energy-yielding substrates on coronary circulation, myocardial oxygen metabolism, and intramyocytic adenylates of perfused Wistar control rat(WC) and spontaneously hypertensive rat(SHR) hearts were examined under basal and $\beta$-adrenergic stimulation conditions. The perfusion medium (1.0mM Ca2+) contained 5mM glucose (+5U/l insulin) in combination with 5mM pyruvate, 5mM lacate, 5mM acetate, or 5mM octanoate as energy substrates. Hearts were perfused with each substrate buffer for 20min under basal conditions. Coronary functinal hyperemia was induced by infusing for 20min isoproterenol (ISO, 1uM), a $\beta$-receptor agonist. Cardiac adenylates, glycolytic intermediates, and coronary venous lactate were measured by using an enzymatic analysis technique. Under basal conditions, acetate and octanoate significantly increased coronary flow(CF) of WC in parallel with myocardial oxygen consumption. However, CF of SHR was partly attenuated by coronary vasoconstriction despite metabolic acidosis. In addition, pyruvate and lactate depressd ISO-induced coronary functional hyperemia in SHR. It should be noted that octanoate exhibited coronary dysfunction under ISO conditions. On the other hand, fat substrates depleted myocardial high energy phosphate pool and accumulated breakdown intermediates. In SHR with coronary vasoconstriction under basal conditions, and with depressed coronary functional hyperemia, high energy phosphates were greatly depleted. These results suggest that energy substrates in the myocardium and coronary smooth muscle alter remarkably coronary circulation, and that coronary circulatory function is associated with a reserve of high energy phosphates and a balance between breakdown and nono synthesis of energy phosphates. These findings could be explained by alterations in the cytosolic redox state manipulated by LDH and hence in the cytosolic phosphorylation potential, which might be involved in hypertension of SHR.

  • PDF