우리나라 여러 해양환경 지역으로부터 확보한 370주의 해양세균, 균류, 미세조류로부터 기초생 리활성(항산화, 항염, 항균, 항암, 항바이러스)을 조사하여 채집지, 분리원, 종(種) 수준에서의 활성결과를 비교하였다. 해양세균의 경우, 일반적으로 유용성이 많이 알려진 Streptomyces 속 과 Bacillus 속에 속하는 균주들이 두드러진 강한 효능이 관찰되었고, 주로 해양퇴적물로부터 유용한 자원을 분리할 수 있었다. 해양균류와 미세조류의 경우에도 종 특이적으로 활성이 강 하게 나타나는 결과를 확인할 수 있었고, 효능 특이적으로 활성을 보이는 결과도 얻을 수 있었 다. 이러한 결과를 바탕으로 추후 특정질병에 선택적으로 효능을 보이는 화학물질 연구 또는 자원 기반 연구 수행 시 유용성을 전제로 한 자원 확보 전략 수립과 산업 활성화를 위한 전 략소재로 우선적 접근이 용이할 수 있는 연구결과라 생각된다. 또한, 이들 결과를 해양바이오 뱅크를 통한 분양소재로 활용함으로써 학계, 산업계에서 활용하여 해양바이오산업 활성화에 좀 더 빠른 접근을 도울 수 있다고 생각한다.
L-asparaginase (ASNase) is a therapeutic enzyme used to treat acute lymphoblastic leukemia. Currently, the most widely used ASNases are originated from bacteria. However, owing to the adverse effects of bacterial ASNases, new resources for ASNase production should be explored. Fungal enzymes are considered efficient and compatible resources of natural products for diverse applications. In particular, fungal species belonging to the genus Trichoderma are well-known producers of several commercial enzymes including cellulase, chitinase, and xylanase. However, enzyme production by marine-derived Trichoderma spp. remains to be elucidated. While screening for extracellular ASNase-producing fungi from marine environments, we found four strains showing extracellular ASNase activity. Based on the morphological and phylogenetic analyses using sequences of translation elongation factor 1-alpha (tef1α), the Trichoderma isolates were identified as T. afroharzianum, T. asperellem, T. citrinoviride, and Trichoderma sp. 1. All four strains showed different ASNase activities depending on the carbon sources. T. asperellem MABIK FU00000795 showed the highest ASNase value with lactose as a carbon source. Based on our findings, we propose that marine-derived Trichoderma spp. are potential candidates for novel ASNase production.
Multiple starters consisting of two Bacillus amyloliquefaciens strains (MJ1-4 and EMD17), Pichiafarinosa SY80, and Rhizopus oryzae were used for Doenjang making. Bacillus strains were selected based on their abilities to inhibit toxinogenic fungi and Bacillus cereus, fibrinolytic activity, and their ability to confer good flavor to Cheonggukjang. P. farinosa SY80 and R. oryzae, previously isolated from soy sauce, were selected because they were not inhibited by two bacilli. Doenjang was prepared by inoculation of multiple starters (A1 Doenjang). Control Doenjang was prepared by inoculation of B. subtilis KACC 16750 (Natto strain) and Aspergillus oryzae KCCM 60166 (A2 Doenjang). Another control (A3 Doenjang) was prepared by inoculation of microorganisms present in rice straw. Doenjang samples were fermented for 70 days at 20℃. pH of 3 samples decreased from the initial value of 6.4 to 5.8~6.0 and titratable acidity (TA) increased from 0.6 to 1.1~1.3. The amount of amino-type nitrogen increased during fermentation. There were slight differences in moisture, crude-protein, and crude-fat contents after 70 days. Contamination of fungi was observed only in A3 Doenjang and B. cereus was not detected from all 3 samples. A1 Doenjang showed the highest fibrinolytic activity and A2 Doenjang the second. These results indicate that Doenjang made with carefully selected starters was functionally improved and microbially more safe.
The purpose of this study was to investigate the effects of swiss ball exercise and taping therapy on back muscle strength on normal college students. The aim of this study was to find effective method for back muscle strengthening. Subjects of 30 college students divided 3 groups(taping therapy group: 10, swiss ball group: 10, control group: 10). All subjects inquired physical conditions and normal exercise habits for data base. Back muscle strength measured before and after 3 weeks intervention. Taping therapy was displayed stable a growth curve in continuative a growth graph of back muscle strength better than swiss ball exercise, because it was taping therapy by periodic effect. The result of this study known to effective either taping therapy or swiss ball exercise, but both taping therapy and swiss ball exercise were effect to increase in back muscle strength.
A MnSOD gene was cloned from the fall webworm, H. cunea. The MnSOD cDNAs encode precursor proteins of 215 amino acid residues. The deduced amino acid sequences of the H. cunea MnSOD cDNA showed 76% identity to B. mori MnSOD and 62-56% to MnSOD sequences from other organisms. MnSOD and Cu/ZnSOD in H. cunea is expressed from all tissues. MnSOD expression is changed at a trace level in infected larvae, while Cu/ZnSOD expression is strongly changed against bacteria, and fungi. The expression level of Cu/ZnSOD increased by different artificial photoperiod (24L:0D), UV irradiation (312nm), and starvation condition, while the expression level of MnSOD only increased by starvation condition. Also, expression of MnSOD and Cu/ZnSOD showed no significant change in 0L:24D condition. In addition to expression levels of Cu/ZnSOD in H. cunea significantly increased by temperature stress and injection with paraquat, but reduced by injection with 10% H2O2. The expression level of MnSOD significantly increased by temperature stress and reduced by injection with 10% H2O2 and paraquat.
Apolipophorin-III (apoLp-III) is a hemolymph protein whose function is to facilitate lipid transport in an aqueous medium in insects. Recently, apolipophorin-III in Galleria mellonella and Hyphantria cunea was shown to play an unexpected role in insect immune activation. We show here a novel possible function/role of the apoLp-III in insects. To investigate the genes which have a relationship with apoLp-III in fall webworm larvae, we reduced endogenous Hc apoLp-III mRNA levels in larvae via RNA interference (RNAi). The RNAi-mediated Hc apoLp-III reduction resulted in the reduction of antioxidants, like MnSOD, catalase, and glutathione S transferase as well as immune proteins. In particular, expression of MnSOD commonly decreased in fat body, midgut, and hemocytes following the knockdown of Hc apoLp-III, which induced an elevated level of superoxide anion in Hyphantria cunea larvae. The observed effect of Hc apoLp-III RNAi suggests that Hc apoLp-III is related to the action/expression of antioxidants, especially MnSOD.
A new insect member of the STAT family of transcription factors (HcSTAT) has been cloned from the lepidopteran, Hyphantria cunea. The domain involved in DNA interaction and the SH2 domain are well conserved. The gene is transcribed at a low level during all stages of development, and transcribed in hemocyte, fat body, midgut, epidermis, and Malpighian tubule. Especially, hemocyte and Malpighian tubule showed transcriptional activation of HcSTAT upon Gram-negative and -positive bacteria challenge. Gram-negative and -positive bacteria challenge specifically results in nuclear translocation of HcSTAT protein and induction of DNA-binding activity that recognizes a STAT target site in H. cunea hemocyte. In vivo treatment with sodium orthovanadatetranslocates HcSTAT to the nucleus in hemocyte cells.
Reactive oxygen species (ROS) is toxic to living organisms, because its high reactivity causes oxidative damage to proteins, nucleic acids, and lipids. Superoxide dismutase (SOD) is an enzyme facilitating the removal of superoxide anions from living organisms. This study focused on the cloning of MnSOD cDNA from Hyphantria cunea and its induction upon bacterial infection and various stresses. The open reading frame of MnSOD is composed of 645 bp, encoding 215 amino acid residues. The theoretical molecular mass and pI of putative MnSOD was evaluated to be 24276 Da and 9.14, respectively. The MnSOD from H. cunea is highly similar to human MnSOD (59.5%) as well as Bombyx mori MnSOD (76.2%). MnSOD showed no big induction upon bacterial infection and stresses, compared to that of Cu/ZnSOD.
Apolipophorin-Ⅲ (apoLp-Ⅲ) is a hemolymph protein whose function is to facilitate lipid transport in an aqueous medium in insect. Recently, apolipophorin-Ⅲ in Galleria mellonella and Hyphantria cunea was shown to play an unexpected role in insect immune activation. We show here a novel possible function/role of apoLp-Ⅲ in insects. To investigate the genes which have a relationship with apoLp-Ⅲ in fall webworm larvae, we reduction of endogenous Hc apoLp-Ⅲ mRNA levels in larvae via RNA interference (RNAi). The RNAi-mediated Hc apoLp-Ⅲ reduction resulted in the reduction of antioxidants, like MnSOD, catalase, and glutathione S transferase as well as immune proteins. In particular, expression of MnSOD commonly decreased in fat body, midgut, and hemocytes following the knockdown of Hc apoLp-Ⅲ, which induced an elevated level of superoxide anion in H. cunea larvae. The observed effect of Hc apoLp-Ⅲ RNAi suggests that Hc apoLp-Ⅲ is related to the action/expression of antioxidants.
Innate immunity responses are triggered by the immune challenge and therefore involve signaling processes. The cellular response is initiated by hemocytes and mainly involves phagocytosis and encapsulation of intruders by these cells. To address whether Hc-STAT is activated upon bacterial challenge, we examined the subcellular location of STAT protein in hemocyte by immunostaining. A new insect member of the STAT family of transcription factors (Hc-STAT) has been cloned from the lepidopteran, Hyphantria cunea. The domain involved in DNA interaction and the SH2 domain are well conserved. The gene is transcribed at a low level during all stages of development, and the protein is present in hemocytes, fat body, midgut, epidermis, and Malphigian tuble (Mt). Especially, hemocytes and Mt showed transcriptional activation of Hc-STAT upon Gram (-) bacteria and fungal challenge. Gram (-) bacteria and fungal challenge specifically results in nuclear translocation of Hc-STAT protein and induction of DNA-binding activity that recognizes a STAT target site in H. cunea hemocyte. In vitro treatment with pervanadate translocates Hc-STAT to the nucleus in hemocyte cells. Here we report the first evidence for the involvement hemocyte JAK/STAT pathway upon microbial infection in lepidopteran insect.
Reactive oxygen species (ROS) is toxic to living organisms, because its high reactivity causes oxidative damage to proteins, nucleic acids, and lipids. Superoxide dismutase (SOD) is an enzyme facilitating the removal of superoxide anions from living organisms. This study focused on the cloning of MnSOD cDNA from Hyphantria cuneaand its induction upon bacterial infection and various stresses. The open reading frame of MnSOD is composed of 645 bp, encoding 215 amino acid residues. The theoretical molecular mass and pI of putative MnSOD was evaluated to be 24276 Da and 9.14, respectively. The MnSOD from H. cunea is highly similar to human MnSOD (59.5%) as well as Bombyx mori MnSOD (76.2%). MnSOD showed no big induction upon bacterial infection and stresses, compared to that of Cu/ZnSOD.
Innate immunity responses are triggered by the immune challenge and therefore involve signaling processes. The cellular response is initiated by hemocytes and mainly involves phagocytosis and encapsulation of intruders by these cells. To address whether Hc-STAT is activated upon bacterial challenge, we examined the subcellular location of STAT protein in hemocyte by immunostaining. A new insect member of the STAT family of transcription factors (Hc-STAT) has been cloned from the lepidopteran, Hyphantria cunea. The domain involved in DNA interaction and the SH2 domain are well conserved. The gene is transcribed at a low level during all stages of development, and the protein is present in hemocytes, fat body, midgut, epidermis, and Malphigian tuble (Mt). Especially, hemocytes and Mt showed transcriptional activation of Hc-STAT upon Gram (-) bacteria and fungal challenge. Gram (-) bacteria and fungal challenge specifically results in nuclear translocation of Hc-STAT protein and induction of DNA-binding activity that recognizes a STAT target site in H. cunea hemocyte. In vitro treatment with pervanadate translocates Hc-STAT to the nucleus in hemocyte cells. Here we report the first evidence for the involvement hemocyte JAK/STAT pathway upon microbial infection in lepidopteran insect.