In this study, Pleurotus ostreatus No.42 was cultured in glucose-peptone-yeast-wheat bran medium using a previously reported novel rotary draft tube bioreactor. Versatile peroxidase (VP), a lignin-degrading enzyme, was isolated from a pellet-type mycelium culture grown in the medium for seven days. The VP was purified by sequentially applying ultra-filtration, DEAESepharose CL-6B column, and Mono Q column. SDS-PAGE analysis revealed the molecular weight of VP to be 36.4 KDa with an isoelectric point of 3.65. The amino acid sequence was confirmed as VTCATGQTT. The purified VP was observed to possess the property of not only oxidizing Mn ions but also decomposing veratryl alcohol, a non-phenolic compound. The catalytic ability of VP is a subject for future research.

Expression changes of stress-induced peroxidase (swpa2 and swpa4) and storage root-specific sporamin (spo-A and spo-B) genes were examined using qRT-PCR after treatment with wounding and bacterial pathogens on leaves of sweetpotato (Ipomoea batatas) plants. As a result of examining the expression change in the wounding treatment condition for 48 hours after treatment, which is a representative physical stress, the expression of all genes increased after 12 hours of wounding treatment, but the expression pattern of each gene group showed distinct differences thereafter. Expression levels of swpa2 and swpa4 strongly increased up to 36 or 48 hours after wounding treatment, whereas spo-A and spo-B expression levels strongly decreased after 24 or 36 hours after wounding treatment. Peroxidase and sporamin genes are involved in the early response after wounding treatment and, in particular, the peroxidase swpa2 and swpa4 genes are also involved in the late response after wounding treatment. Gene expression analysis after infection with P. chrysanthemi, which causes softness in sweetpotato, showed that the swpa2 and swpa4 genes were weakly induced after 8 hours and then strongly induced after 20 hours during pathogen infection. Expression of the spo-A gene was weakly induced in the pathogen-treated group after 20 hours, whereas spo-B showed an expression pattern similar to that of the peroxidase genes. The above results indicate that expression of the stress-induced peroxidase gene used in this study is induced not only by abiotic stress but also by biological stress caused by bacterial pathogen invasion and that peroxidase plays an important function in the initial defense response.

리그닌 분해효소의 생산은 나선형 리본이 있는 새로운 형태의 회전식 통풍관 생물 반응기(RTB)를 사용하여 느타리(Pleurotus ostreatus) No.42에 의해 실시하였다. 락게 이즈(laccase)의 최대 생산량은 배양 3일 후 약 8,200 U/ bioreactor 수준에 도달한 후 감소하였다. 반면에, 망간퍼 옥시데이즈(MnP)의 최대 생산은 6일 배양 후 약 8,400 U/bioreactor의 수준에 도달하였다. 그러나 이 발효조에서 리그닌퍼옥시데이즈(LiP)는 검출되지 않았다. 그 결과 회전식 통풍관 생물 반응기(RTB)가 리그닌 분해효소를 대규모 생산을 위해 성공적으로 생산할 수 있음을 보여주었다. 이 발효기에서 망간퍼옥시데이즈의 정제 과정은 Sepharose CL-6B, Superdex 75 prep grade 및 Mono-Q에 대한 크로마토그래피를 포함하여 정제하였다. 이 주요 효소는 sodium dodecyl sulfate-polyacrylamide 겔 전기영동(SDS-PAGE)에서 분자량 36,400, pI 3.95의 등전점(IEF)으로 각각 확인되었다. 이 발효기의 주요 효소 N-말단 서열은 정치 및 진탕배양과 같은 다른 배양조건에서 보고된 MnP3 효소와 동일하였다.

리그닌 분해 담자균류로 알려져 있는 느타리버섯균 No.42는 망간퍼옥시데이즈(MnP) 및 락게이즈(Lac)를 생산하였으나 글루코오스-펩톤-이스트-밀기울(GPYW)배지를 이용한 정치배양조건하에서 리그닌퍼옥시데이즈(LiP)활성은 검출 되지 않았다. 한편, 동일배지에서 망간퍼옥시데이즈 활성은 11일째 80(3.6) U/flask(ml)의 최대 생산되었다. 망간퍼옥시 데이즈 분리정제는 Sepha-ros CL-6B 및 Mono-Q 컬럼순으로 수행하였으며 주요 망간퍼옥시데이즈 isozyme은 단일밴 드로 분자량은 36.4KDa이였다. N-말단으로부터 19개의 아 미노산 배열은 단백질 자동 분석 장치로 분석한 결과 ATCADGRTTANAACCVLFP를 나타내었다. 느타리버섯균 No.42의 정치배양조건 하에서 세포외로 생산되는 중요한 망간퍼옥시데이즈 isozyme의 N-말단 아미노산 배열은 이전에 보고된 MnP3의 아미노산 배열과 동일하였다.

Carbon-based magnetic nanostructures in several instances have resulted in improved physicochemical and catalytic properties when compared to multi-wall carbon nanotubes (MWCNTs) and magnetic nanoparticles. In this study, magnetic MWCNTs with a structure of NixZnxFe2O4/MWCNT as peroxidase mimics were fabricated by the one-pot hydrothermal method. The structure, composition and morphology of the nanocomposites were characterized with X-ray diffraction (XRD), Fourier transform infrared spectroscopy and transmission electron microscopy. The magnetic properties were investigated with a vibrating sample magnetometer. The peroxidase-like catalytic activity of the nanocomposites was investigated by colorimetric and electrochemical tests with 3,3´,5,5´-tetramethylbenzidine (TMB) and H2O2 as the substrates. The results show that the synthesis of the nanocomposites was successfully performed. XRD analysis confirmed the crystalline structures of the NixZnxFe2O4/ MWCNT nanohybrids and MWCNTs. The main peaks of the NixZnxFe2O4/MWCNTs crystals were presented. The Ni0.25Zn0.25Fe2O4/MWCNT and Ni0.5Zn0.5Fe2O4/MWCNT nanocatalysts showed nearly similar physicochemical properties, but the Ni0.5Zn0.5Fe2O4/MWCNT nanocatalyst was more appropriate than the Ni0.25Zn0.25Fe2O4/MWCNT nanocatalyst in terms of the magnetic properties and catalytic activity. The optimum peroxidase-like activity of the nanocatalysts was obtained at pH 3.0. The Ni0.5Zn0.5Fe2O4/MWCNT nanocatalyst exhibited a good peroxidase-like activity. These magnetic nanocatalysts can be suitable candidates for future enzyme-based applications such as the detection of glucose and H2O2.

주스의 품질악화를 초래하는 효소를 불활성화시키기 위해서 감귤 주스와 사과 주스 추출물에 초고압 처리와 열처리를 하여 각 시료의 PPO와 POD의 불활성화 정도를 확인하고 동역학 모델에 적용하여 이들의 불활성화 특성을 연구하였다. 감귤 주스와 사과 주스에서 모두 PPO가 POD보다 열과 압력에 대해서 상대적으로 내성을 갖는 것을 알 수 있었으며, 초고압 처리 과정을 거쳤을 때 보다는 열처리 과정을 거쳤을 때 PPO와 POD가 효과적으로 불활성화되는 것을 확인할 수 있었다. 초고압 기술은 처리 후에도 제품의 영양분과 색의 변화에 영향을 미치지 않기에, 최상의 상태와 품질로 제품을 제공 할 수 있는 장점을 가지고 있지만, 효소를 불활성시키기에는 부족하다는 단점을 가지고 있다. 이를 보완하여 산업적으로 이용되어지기 위해서는 영양분이 파괴되지 않는 열 처리 조건을 잡고, 효소를 효과적으로 불활성시킬 수 있는 초고압 처리 조건과 결합하여 최상의 결과를 얻을 수 있는 조건을 찾는 연구가 필요하다고 판단된다.

Glutathione (GSH)은 직접적으로 활성산소종을 제거하거나 GSH peroxidase와 같은 활성산소종 제거 효소의 조효소로써, 산화적 스트레스로부터 세포를 방어하는 데 중요한 역할을 한다. GSH peroxidase는 두 분자의 GSH을 이용해 세포 내 과산화수소를 물로 전환한다. N-acetyl-L cysteine (NAC)는 항산화제 중 하나로 세포 내 GSH의 전구물질로 이용된다. 본 연구는, 0mM에서 20mM의 NAC 단독 처리

Plasma glutathione peroxidase (pGPx) is an extracellular antioxidative selenoenzyme which has been detected in various adult tissues, but little is known about the expression and distribution of pGPx during embryogenesis. To investigate the expression patterns of pGPx during embryogenesis, we performed quantitative real-time PCR, in situ hybridization, Western blot, and immunohistochemistry analyses in whole embryos or each developing organ of mice on embryonic days (E)7.5–18.5. In whole embryos of E7.5–8.5, pGPx mRNA was more typically expressed in extra-embryonic tissues including ectoplacental cone, trophectoderm, and decidual cells than in embryos. However, after E9.5, pGPx mRNA and protein levels were increased in the embryos with differentiation and growth, but trended to gradually decrease in the extra-embryonic tissues until E18.5. In sectioned embryonic tissues on E13.5–18.5, pGPx mRNA and protein were mainly expressed in the developing nervous tissues, the sensory organs, and the epithelia of lung, skin, and intestine, the heart and artery, and the kidney. In particular, pGPx immunoreactivity was very strong in the developing liver. These results indicate that pGPx is spatio-temporally expressed in various embryonic organs as well as extra-embryonic tissues, suggesting that pGPx may function to protect the embryos against endogenous and exogenous reactive oxygen species during organogenesis.

Phospholipid hydroperoxide glutathione peroxidase (PHGPx) is a unique antioxidant enzyme involved in reduction of peroxidized phospholipids within biomembranes. To investigate the expression pattern of the PHGPx gene during fetal development, in situ hybridization analyses were performed using mouse FITC-labeled PHGPx cRNA probes in fetal tissues on embryonic days (Ed) 13.5-18.5. During these periods, PHGPx mRNA appeared in the developing telencephalon, diencephalon, spinal cord, and spinal ganglion. In particular, PHGPx mRNA was strongly expressed in pyramidal cells of the cerebral cortex. On Eds 17.5-18.5, PHGPx mRNA was detected in various tissues including liver, intestinal villi and crypt, pancreas, lung, and olfactory epithelium of the nasal cavity. In addition, PHGPx mRNA was highly expressed in the inner ear on Eds 14.5-18.5, brown fat on Ed 17.5, and adrenal gland on Ed 18.5. It is conceivable that PHGPx may act as an important antioxidant against fetal oxidative stress during mouse organogenesis.

Phospholipid-hydroperoxide glutathione peroxide (PHGPx) is an antioxidant enzyme that reduces lipid hydroperoxides in biomembranes. Here, we cloned and characterized cys-PHGPx from the bumblebee Bombus ignitus (Bi-PHGPx). The Bi-PHGPx gene consists of 4 exons, encoding 168 amino acid residues with a canonical cys-codon at residue 45 and active site residues Gln82 and Trp134. Recombinant Bi-PHGPx, expressed as a 19 kDa protein in baculovirus-infected insect cells, exhibited enzymatic activity against PLPC-OOH and H2O2 using glutathione as an electron donor. Tissue distribution analyses showed the presence of Bi-PHGPx in all tissues examined. Bi-PHGPx transcripts were upregulated by stresses, such as wounding, H2O2 exposure, external temperature shock, and starvation. Under H2O2 overload, the RNA interference (RNAi)-induced thioredoxin peroxidase (BiTPx1)-knock-down B. ignitus worker bees showed upregulated expression of Bi-PHGPx in the fat body. These results indicate that Bi-PHGPx is a stress-inducible antioxidant enzyme that acts on phospholipid hydroperoxide and H2O2.

A thioredoxin peroxidase (TPx) gene was cloned from the bumblebee, Bumbus ignitus. The B. ignitus TPx (BiTPx) contains an open reading frame of 585 bp encoding 195 amino acid residues and possesses two cysteine residues that are characteristic of 2-Cys subgroup of peroxiredoxin family. The deduced amino acid sequence of the BiTPx cDNA showed 90% identity to Apis melifera (AmTPx-1), 80% to Aedes aegypti (AaTPx), and 78% - 47% to other insect 2-Cys TPx. Northern blot analysis revealed the presence of BiTPx transcripts in all tissues examined. Western blot analysis showed the presence of the BiTPx in the fat body, midgut, muscle and epidermis, but not in the hemolymph, suggesting the BiTPx is not secretable. The cDNA encoding BiTPx was expressed as a 27-kDa polypeptide in baculovirus-infected insect Sf9 cells. The purified recombinant BiTPx was shown to reduce H2O2 in the presence of electrons donated by dithiothreitol and shown to be active in the presence of thioredoxin as electron donor.

감국(Dendranthema indicum)의 환경조건에 따른 개화기의 초장, 개화소요일수, 카로티노이드 함량 및 peroxidase 활성을 고랭지(대관령, 표고 800m)와 평난 지(강릉, 표고 20m)의 노지와 온실에서 각각 비교하여 원예화 개발 가능성을 검토하였다. 환경조건에 따른 광 량은 고랭지 노지 > 평난지 노지 > 고랭지 온실 > 평난지 온실 순으로 감소하였다. 고랭지의 노지에서는 UV A와 UV B의 세기가 22.4, 3.02W·m-2로 평난지 노지의 19.9, 2.24W·m-2보다 각각 12.6, 34.8% 더 강했다. 감국의 초장은 평난지의 온실조건에서 가장 컸 으나 개화는 고랭지의 노지에서 가장 빨랐다. 감국 꽃 의 주요 색소인 카로티노이드는 고랭지의 온실에서 60.9OD·g-1으로 가장 많았다. 그러나 peroxidase 활성 은 평난지의 온실에서 자란 식물체에서 5.72unit·mg-1 protein 가장 높았다.

본 연구는 정자 내의 phsopholipid hydroxide glutathion peroxidase (PHGPx) mRNA 발현 수준, heparin-binding protein (HBP) mRNA 발현 수준, 수태율 그리고 산자수 사이의 관계를 조사하고자 실시하였다. 수태율과 산자수 사이에 있어서 상관관계는 나타나지 않았다. PHGPx mRNA 발현 수준에 있어 산자수가 10두 이상 군에서 (2,414.7±400.7) 8두 미만 군보다 (1,875.8±311.2) 높게 나타났으나 유의적인 차이는 없었다. HBP mRNA 발현 수준에 있어서도 산자수 10두 이상 군에서 (2,255.9±360.8) 8두 미만 군보다 (2,155.4±378.0) 약간 높은 결과를 보였으나, 유의적인 차이는 인정되지 않았다. PHGPx mRNA 발현 수준과 산자수 사이의 관계는 (r=0.206) 정의 상관관계를 보였으나, 유의한 상관관계는 나타나지 않았다. 본 실험의 결과, PHGPx와 HBP의 발현수준이 수태율, 산자수와 유의한 상관관계를 보이지 않았기 때문에 PHGPx와 HBP가 정자의 수정능력을 예측하기에는 미흡한 면이 있다.