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The Effect of Platelet Derived Growth Factor - BB Loaded Chitosan/Calcium Metaphosphate on Bone Regeneration (혈소판유래성장인자를 함유한 Chitosan/Calcium Metaphosphate의 골조직재생효과에 관한 연구)

  • Lee, Seung-Yeol;Seol, Yang-Jo;Lee, Yong-Moo;Lee, Ju-Yeon;Lee, Seung-Jin;Kim, Suk-Young;Ku, Young;Rhyu, In-Chul;Han, Soo-Boo;Choi, Sang-Mook;Chung, Chong-Pyoung
    • Journal of Periodontal and Implant Science
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    • v.31 no.1
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    • pp.1-23
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    • 2001
  • Chitosan is biodegradable natural polymer that has been demonstrated its ability to improve wound healing, and calcium metaphosphate(CMP) is a unique class of phosphate minerals having a polymeric structure. In this study, chitosan/CMP and platelet derived growth factor(PDGF-BB) loaded chitosan/CMP sponges were developed, and the effect of the sponges on bone regeneration and their possibility as scaffolds for bone formation by three-dimensional osteoblast culture were examined. PDGF-BB loaded chitosan/CMP sponges were prepared by freeze-drying of a mixture of chitosan solution and CMP powder, and soaking in a PDGF-BB solution. Fabricated sponge retained its 3-dimensional porous structure with $100-200\;{\mu}m$ pores. The release kinetics of PDGF-BB loaded onto the sponge were measured in vitro with $^{125}I-labeled$ PDGF-BB. In order to examine their possibility as scaffolds for bone formation, fetal rat calvarial osteoblastic cells were isolated, cultured, and seeded into the sponges. The cell-sponge constructs were cultured for 28 days. Cell proliferation, alkaline phosphatase activity were measured at 1, 7, 14 and 28 days, and histologic examination was performed. In order to examine the effect on the healing of bone defect, the sponges were implanted into rat calvarial defects. Rats were sacrificed 2 and 4 weeks after implantation and histologic and histomorphometrical examination were performed. An effective therapeutic concentration of PDGF-BB following a high initial burst release was maintained throughout the examination period. PDGF-BB loaded chitosan/CMP sponges supported the proliferation of seeded osteoblastic cells as well as their differentiation as indicated by high alkaline phosphatase activities. Histologic findings indicated that seeded osteoblastic cells well attached to sponge matrices and proliferated in a multi-layer fashion. In the experiments of implantation in rat calvarial defects, histologic and histomorphometric examination revealed that chitosan/CMP sponge promoted osseous healing as compared to controls. PDGF-BB loaded chitosan/CMP sponge further echanced bone regeneration. These results suggested that PDGF-BB loaded chitosan/CMP sponge was a feasable scaffolding material to grow osteoblast in a three-dimentional structure for transplantation into a site for bone regeneration.

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Intra-laboratory Validation of an HPLC Post-column Oxidation Method for the Analysis of PSP Toxins in Oysters and Mussels (굴과 진주담치 중 마비성 패류독소 분석을 위한 HPLC post-column oxidation method의 시험소 내 유효성 검증)

  • Song, Ki Cheol;Lee, Ka-Jeong;Yu, Hong-Sik;Mok, Jong-Soo;Kim, Ji Hoe;Lim, Keun-Sik;Lee, Mi-Ae;Kim, Mee-Hye
    • Korean Journal of Food Science and Technology
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    • v.45 no.2
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    • pp.241-247
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    • 2013
  • AOAC Mouse Bioassay Analysis (MBA) has been the gold standard for the analysis of paralytic shellfish poisoning toxin (PSP toxin) for more than 50 years. However, this method has inaccurate limit of quantification and cannot be used to determine toxic profiles. An HPLC method (PCOX) was optimized for Korean shellfish to establish an alternative or supplementary method for PSP analysis and was intended to be used for the official monitoring and regulation of food. The recovery rate of the PCOX method was 83.5-112.1% and the limit of quantification for total toxin was about $8.6{\mu}g$/100 g. A long-term comparison study showed a good correlation of the PCOX results with the AOAC MBA results: the correlation factors were 0.9534 and 0.9109 for oyster and mussel matrices, respectively. The PCOX method may be used as an alternative or supplementary method for AOAC MBA to monitor the occurrence of PSP and to analyze PSP toxin profile in oysters and mussels.

Optimal supervised LSA method using selective feature dimension reduction (선택적 자질 차원 축소를 이용한 최적의 지도적 LSA 방법)

  • Kim, Jung-Ho;Kim, Myung-Kyu;Cha, Myung-Hoon;In, Joo-Ho;Chae, Soo-Hoan
    • Science of Emotion and Sensibility
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    • v.13 no.1
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    • pp.47-60
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    • 2010
  • Most of the researches about classification usually have used kNN(k-Nearest Neighbor), SVM(Support Vector Machine), which are known as learn-based model, and Bayesian classifier, NNA(Neural Network Algorithm), which are known as statistics-based methods. However, there are some limitations of space and time when classifying so many web pages in recent internet. Moreover, most studies of classification are using uni-gram feature representation which is not good to represent real meaning of words. In case of Korean web page classification, there are some problems because of korean words property that the words have multiple meanings(polysemy). For these reasons, LSA(Latent Semantic Analysis) is proposed to classify well in these environment(large data set and words' polysemy). LSA uses SVD(Singular Value Decomposition) which decomposes the original term-document matrix to three different matrices and reduces their dimension. From this SVD's work, it is possible to create new low-level semantic space for representing vectors, which can make classification efficient and analyze latent meaning of words or document(or web pages). Although LSA is good at classification, it has some drawbacks in classification. As SVD reduces dimensions of matrix and creates new semantic space, it doesn't consider which dimensions discriminate vectors well but it does consider which dimensions represent vectors well. It is a reason why LSA doesn't improve performance of classification as expectation. In this paper, we propose new LSA which selects optimal dimensions to discriminate and represent vectors well as minimizing drawbacks and improving performance. This method that we propose shows better and more stable performance than other LSAs' in low-dimension space. In addition, we derive more improvement in classification as creating and selecting features by reducing stopwords and weighting specific values to them statistically.

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Anti-calcification Effects in Decellularized and Variously Fixed Bovine Pericardium (소심낭 절편의 무세포화와 알코올 전처치를 포함한 여러 고정 처리법 시행 후 석회화 경감 효과 관찰 연구)

  • Hwang, Seong-Wook;Kim, Yong-Jin;Kim, Soo-Hwan;Choi, Seung-Hwa
    • Journal of Chest Surgery
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    • v.43 no.3
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    • pp.235-245
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    • 2010
  • Background: Our goal was to evaluate anti-calcification effects of decellularization and diverse fixing methods including preincubation of the bovine pericardium with ethanol. We also assessed changes in mechanical properties. Material and Method: Harvested bovine pericardium was decellularized with 0.25% sodim dodecysulfate and then treated with 5 methods of fixation: (1) 0.5% glutaraldehyde (GA) for 14 days, (2) 0.5% GA for 5 days, 2% GA for 2 days and 0.25% GA for 7 days, (3) 0.5% GA for 5 days, 2% GA for 2 days, 0.25% GA for 7 days, and then 70% ethanol for 2 days, (4) 0.5% GA for 5 days, a mixture of 2% GA and 70% ethanol for 2 days, and 0.25% GA for 7 days, (5) 0.5% GA for 5 days, a mixture of 2% GA, 65% ethanol, and 5% octanediol for 2 days and then 0.25% GA for 7 days. All treated bovine pericardia were tested for histological variables, lipid content, and mechanical properties including tensile strength and thermal stability. A total 10 kinds of differently treated bovine pericardia were implanted into rat subdermis and harvested 8 weeks later. Harvested pericardia were evaluated for calcium content. Result: No protein denaturation was observed microscopically after decellularization. There was a 32% mean decrease in tensile strength index after decellularization in the bovine pericardium group fixed. Octanediol preincubation attenuated the decrease in tensile strength and maintained thermal stability. TG and cholesterol were not affected by decellularization but were decreased by organic solvent. Calcium content was decreased after decellularization, and organic solvent preincubation decreased calcification in the non-decellularized bovine pericardium group. Conclusion: Decellularization and organic solvent preincubation have anti-calcification effects but decellularization may cause mechanical instability. A method of decellularization and fixation that does not cause damage to matrices will be needed for evaluation of the next step in using tissue-engineering for replacement of cardiac valves.

Validation of Trienzyme Extraction-Microplate Assay for Folate in Korean Ancestral Rite Food (Trienzyme Extraction-Microplate Assay를 이용한 한국 차례 및 제사 음식의 엽산 분석 및 검증)

  • Park, Su-Jin;Jeong, Beom-Gyun;Jung, Jae Eun;Kim, Hyeon-Young;Jung, Gil-Rak;Hwang, Eun-Jung;Yoon, Sung-Won;Hyun, Taisun;Lee, Junsoo;Chun, Jiyeon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.5
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    • pp.716-724
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    • 2015
  • Trienzyme extraction coupled with microplate assay (Lactobacillus casei subsp. rhamnosus) was validated and applied for the determination of folate (vitamin B9) in Korean ancestral rite foods. Foods included five Guk (Tang), eleven Sookchaes, eight Jeoks, nine Jeons, six Jjims, and twenty desserts. Folate was detected in all samples: Guk (Tang) 4.62~18.84, Sookchae 6.13~48.40, Jeok 5.49~49.50, Jeon 6.96~30.77, Jjim 10.34~38.88, and desserts $3.33{\sim}49.55{\mu}g/100g$. The lowest folate content was observed in Sikhye ($3.33{\mu}g/100g$), whereas the highest was observed in Songhwa-dasik ($49.55{\mu}g/100g$). Folate analyses of certified reference materials, BCR-121 (whole meal flour) and BCR-487 (pig liver), showed good recoveries of 90.0% (0.45 mg/kg) and 92.4% (12.3 mg/kg), respectively. The recoveries (96.0 to 106.2%) obtained by analyzing eight spiked samples with different matrices also showed good accuracy. Both repeatability and reproducibility were less than 5%, indicating good precision. The quality control chart (n>30) obtained by running commercial folate fortified-wheat flour once a week for about 10 months showed that all assays were under control. All validation method and analytical quality control results showed that folate contents in Korean ancestral rite foods produced by microplate assay were reliable enough to be used for the construction of a national folate database.

Biological Activities of Calcium Polyphosphate (Calcium polyphosphate의 생물학적 활성도에 관한 연구)

  • Seol, Yang-Jo;Lee, Jae-Il;Lee, Yong-Moo;Lim, Yoon-Tak;Kim, Seok-Young;Ku, Young;Rhyu, In-Chul;Hahm, Byung-Do;Han, Soo-Boo;Choi, Sang-Mook;Chung, Chong-Pyoung
    • Journal of Periodontal and Implant Science
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    • v.30 no.2
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    • pp.213-231
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    • 2000
  • 이 연구의 목적은 다공성의 CPP 내부에 쥐의 장골의 골수에서 유래된 세포를 접종하고 3차 원적으로 배양하여 CPP가 골 형성을 위한 조직공학의 지지체로 적용가능한가를 연구하는 것과 Calcium PolyPhosphate(CPP)의 돌연변이 유발성을 검사하는 것이다. 무수 ($Ca(H_2PO_4)$)를 condensation하여 무결정의 ($Ca(PO_3)$)를 얻고 이를 용융하고 냉각시킨 후 분쇄하여 Calcium polyphosphate(CPP) powder를 얻었다. 다공성의 CPP는 5% $SiO_2$를 첨가하여 sponge 형태로 $450-550{\mu}m$ 소공의 크기를 가지는 것과(CPP-45ppi) $200-300{\mu}m$의 소공의 크기를 가지는 것(CCP-60ppi) 2가지로 제작하였다. 각각의 CPP matrices는 $5mm{\times}5mm{\times}1mm$의 블록 형태로 만들었다. 체중 100g 내외의 백서에서 장골(femur, tibia)을 채취하여 백서의 장골 골수 세포를 분리하여 배양한 후 24well에 CPP block을 넣고 CPP block 당 $10^5$개의 배양한 세포를 접종하였다. 배양 1, 7, 14, 및 21 일째에 각 well에서 trypsin EDTA를 이용하여 2회 반복하여 cell을 분리하였고, 원심분리한 후 hemacytometer로 측정하였다. 또, 45ppi와 60ppi, 그 리 고 Tissue Culture Polystyrene(control group)에 접종, 배양된 세포들의 염기성 인산분해효소활성도를 배양 7, 14, 및 21 일째에 각각 측정하였다. 각 기간별로 배양된 세포-CPP 혼합체내에서 세포의 부착 및 증식과 형성된 조직의 3차원적 형태를 관찰하기 위하여 주사전자현미경하에서의 관찰하였다. CPP의 돌연변이 유발성 검사 (mutagenicity test)를 위해 hypoxanthine-guanine phosphoribosyl transferase(HPRT) assay를 하였다. NIH3T3 cell line과 CHO-K1 cell line으로 각각 $1000{\mu}g/m{\ell}$, $100{\mu}g/m{\ell}$, $10{\mu}g/m{\ell}$ 그리고 $1{\mu}g/m{\ell}$의 CPP 농도에서 측정하였다. 통계적 분석을 위해서 모든 측정은 각군당 4개체 이상 시험하였고, 각 측정값은 평균값${\pm}$표준편차로 나타내었다. 각 군간의 통계적 유의성 검정을 위해서 Analysis of variance(ANOVA)를 이용하였고 Tukey의 방법으로 사후분석을 실시하였다. 제작된 CPP matrices 소공들이 서로간에 연결이 잘 되어있는 형태였다. 두 가지로 제조된 CPP(45ppi와 60ppi) 모두에서 세포의 부착이 잘 일어났고, 부착된 세포의 분열도 잘 일어났다. 2 가지의 CPP 모두에서 7, 14, 21일째의 세포 수는 1일째에 비해 유의성 있게 증가하였다(P<0.01). 3차원적 구조인 Calcium PolyPhosphate에서 배양한 세포는 24well dish(tissue culture polystyrene)에서 평면적으로 배양한 대조군의 세포에서 보다 염기성 인산분해효소 (Alkaline Phosphatase)를 유의성 있게 높게 나타냈다. 주사전자현미경에서 세포-CPP 혼합체를 관찰한 결과, CPP block에 세포들이 잘부착되어 있었고, 시간이 지남에 따라 세포가 여러 층을 형성하면서 뭉치는 현상을 보였다. 또, HPRT assay 결과 , Calcium PolyPhosphate는 돌연변이 유발성을 보이지 않았다. 이상의 결과로 볼 때 CPP에는 세포부착이 잘 일어나고, 지지체 상에서 세포의 분열도 활발하게 일어나므로 골조직을 위한 조직공학의 우수한 지지체가 될 수 있을 것으로 사료된다.

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Estimation of Genetic Parameter for Linear Type Traits in Holstein Dairy Cattle in Korea (Holstein종 젖소의 선형심사형질에 대한 유전모수추정)

  • Lee, Ki-Hwan;Sang, Byung-Chan;Nam, Myoung-Soo;Do, Chang-Hee;Choi, Jae-Gwan;Cho, Kawng-Hyun
    • Journal of Animal Science and Technology
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    • v.51 no.5
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    • pp.345-352
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    • 2009
  • This study utilized 332,625 records of linear type scores consisting for 15 primary traits, 22,175 final score and 84,612 pedigree information of 22,175 Holstein cows from 1993 to 2007 in Korea to estimate genetic parameters for 16 type traits. Genetic and error (co)variances between two traits selected from 16 traits were estimated using bi-trait pairwise analyses with DFREML package. The estimated heritabilities for stature (ST), strength (STR), body depth (BD), dairy form (DF), rump angle (RA), thurl width (TW), rear legs side view (RLSV), foot angle (FA), fore udder attachment (FUA), rear udder height (RUH), rear udder width (RUW), udder cleft (UC), udder depth (UD), front teat placement (FTP), front teat length (FTL) and final score (FS) were 0.31, 0.21, 0.25, 0.10, 0.29, 0.19, 0.09, 0.06, 0.12, 0.13, 0.12, 0.08, 0.26, 0.20, 0.28 and 0.15, respectively. ST had the highest positive genetic correlation with BD (0.90), while RLSV had the highest negative genetic correlation with FA (-0.56). RA had negative genetic correlation with most udder traits (-0.17~-0.02). Especially, RUW had the higher positive genetic correlation with STR (0.60), BD (0.62), and TW (0.49), however, UD had the higher negative genetic correlation with STR (-0.40) and BD (-0.40). FTL had negative genetic correlation with FUA, RUH, RUW, UC and UD. FS had positive genetic correlation with UC, UD and FTP (0.12, 0.18 and 0.20). However, additional research is needed on the use of these parameters in the genetic evaluation because estimated genetic and error variance-covariance matrices were not positive definite.

Clinical Usefulness of PET-MRI in Lymph Node Metastasis Evaluation of Head and Neck Cancer (두경부암 림프절 전이 평가에서 PET-MRI의 임상적 유용성)

  • Kim, Jung-Soo;Lee, Hong-Jae;Kim, Jin-Eui
    • The Korean Journal of Nuclear Medicine Technology
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    • v.18 no.1
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    • pp.26-32
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    • 2014
  • Purpose: As PET-MRI which has excellent soft tissue contrast is developed as integration system, many researches about clinical application are being conducted by comparing with existing display equipments. Because PET-MRI is actively used for head and neck cancer diagnosis in our hospital, lymph node metastasis before the patient's surgery was diagnosed and clinical usefulness of head and neck cancer PET-MRI scan was evaluated using pathological opinions and idiopathy surrounding tissue metastasis evaluation method. Materials and Methods: Targeting 100 head and neck cancer patients in SNUH from January to August in 2013. $^{18}F-FDG$ (5.18 MBq/kg) was intravenous injected and after 60 min of rest, torso (body TIM coil, Vibe-Dixon) and dedication (head-neck TIM coil, UTE, Dotarem injection) scans were conducted using $Bio-graph^{TM}$ mMR 3T (SIEMENS, Munich). Data were reorganized using iterative reconstruction and lymph node metastasis was read with Syngo.Via workstation. Subsequently, pathological observations and diagnosis before-and-after surgery were examined with integrated medical information system (EMR, best-care) in SNUH. Patient's diagnostic information was entered in each category of $2{\times}2$ decision matrix and was classified into true positive (TP), true negative (TN), false positive (FP) and false negative (FN). Based on these classified test results, sensitivity, specificity, accuracy, false negative and false positive rate were calculated. Results: In PET-MRI scan results of head and neck cancer patients, positive and negative cases of lymph node metastasis were 49 and 51 cases respectively and positive and negative lymph node metastasis through before-and-after surgery pathological results were 46 and 54 cases respectively. In both tests, TP which received positive lymph node metastasis were analyzed as 34 cases, FP which received positive lymph node metastasis in PET-MRI scan but received negative lymph node metastasis in pathological test were 4 cases, FN which received negative lymph node metastasis but received positive lymph node metastasis in pathological test was 1 case, and TN which received negative lymph node metastasis in both two tests were 50 cases. Based on these data, sensitivity in PET-MRI scan of head and neck cancer patient was identified to be 97.8%, specificity was 92.5%, accuracy was 95%, FN rate was 2.1% and FP rate was 7.00% respectively. Conclusion: PET-MRI which can apply the acquired functional information using high tissue contrast and various sequences was considered to be useful in determining the weapons before-and-after surgery in head and neck cancer diagnosis or in the evaluation of recurrence and remote detection of metastasis and uncertain idiopathy cervical lymph node metastasis. Additionally, clinical usefulness of PET-MRI through pathological test and integrated diagnosis and follow-up scan was considered to be sufficient as a standard diagnosis scan of head and neck cancer, and additional researches about the development of optimum MR sequence and clinical application are required.

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Evaluation of Residual Pesticides in Commercial Agricultural Products Using LC-MS/MS in Korea (LC-MS/MS를 이용한 한국에서 유통되는 농산물의 잔류 농약 평가)

  • Lee, Hwa-Mi;Hu, Soo Jung;Lee, Hyun-Sook;Park, Seung Young;Kim, Nam-Sun;Shin, Yong Woo;Choi, Keun Hwa;Kim, Sung Il;Nam, Seul Yi;Cho, Dae Hyun
    • Korean Journal of Food Science and Technology
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    • v.45 no.4
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    • pp.391-402
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    • 2013
  • Pesticide residues in agricultural products in Korea were evaluated to create a data base to inform food policy. Pesticide residues were investigated in 17 commodities collected from 9 provinces in 2012. Our analytical method of multiresidue analysis, involved the analysis of individual recovery, linearity, limit of detection, and limit of quantitation of 60 pesticides in six matrices. The pesticides, including acetamiprid and forchlorofenuron, were evaluated in 358 agricultural samples by liquid chromatography/tandem mass spectrometry. Twenty three pesticides were detected from 67 samples. The results showed that the ratio of estimated daily intake to acceptable daily intake was 0.00009-0.0861% by the intake assessment, which means that residual pesticides in agricultural products in Korea are properly controlled.

Improvement of L-Lysine Productivity by Using Cell Fusion and Immobilized System (세포융합과 고정화 시스템을 이용한 L-Lysine의 생산성 향상)

  • Ryu, Beung-Ho;Kim, Hye-Sung;Roh, Myung-Hoon;Park, Bob-Gyu;Chung, Jong-Soon;Bai, Ki-Chul
    • Korean Journal of Food Science and Technology
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    • v.21 no.1
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    • pp.154-163
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    • 1989
  • This studies were designed to improve the productivity of L-lysine by protoplast fusion and immobilized system of fusants using strains of Brevibacterium flavum ATCC 21528, Brevibacterium lactofermentum ATCC 21086 and Corynebacterium glutamicum 820. Mutants were isolated with concentration method of $300{\mu}g/ml$ penicillin-G after treatment of $250{\mu}g/ml$ N-methyl-N-nitro-N-nitrosoguanidine. B. flavum $37-2(Hos^-,\;Kan^r,\;AEC^r)$, B. lactofermentum $6-2(Ile^-,\;Val^-,\;Str^r,\;AEC^r)$ and C. glutamicum 57-5$(Met^-,\;Thr^-,\;Rif^r,\;AEC^r)$ were isolated from mutants. Protoplasts were induced by being incubated with $500{\mu}g/ml$ lysozyme of lysis solution for 6 hr and the ratio of protoplast formation and regeneration were ranging from 97-99% and 33-37%, respectively. Fusion frequencies of fusants of BBFL 21, BCFG 37 and BCLG 59 were shown in the range from $1.25{\times}10^{-6}\;to\;5.83{\times}10^{-7}$ under the optimum conditions. The fusant BBFL 21 showed the highest productivity of $411.1\;ng/ml{\cdot}hr$ L-lysine in the lysine productivity broth at $30^{\circ}C$ for 72hr. In the immobilization systems, fusant BBFL 21 was employed in various polymer matrices such as sodium alginate, polyacrylamide, agar and ${\alpha}-carrageena$. The immobilization of sodium alginate showed the highest productivity of $413\;ng/ml{\cdot}hr$ L-lysine in the batch system. Continuous fermentation of immobilization system by using tube fermentor was produced the highest productivity $416.7\;ng/ml{\cdot}hr $ L-lysine under optimum condition.

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