다시마(Laminaria japonicaus) Alginate의 가열가수분해에 따른 물리${\cdot}$화학적 및 생물학적 특성에 관한 연구 -2. 저분자 alginate의 항돌연변이효과와 Cholesterol, Glucose 및 카드뮴(Cd) 결합능의 변화-

김육용,이근우,김건배,조영제,KIM Yuck-Yong,LEE Keun-Woo,KIM Geon-Bae,CHO Young Je 한국수산과학회 2000 한국수산과학회지 Vol.33 No.5

Alginate의 광범위하고 효율적인 이용을 목적으로 평균분자량이 약 10,000 (HAG-10), 50,000 (HAG-50) 및 100,000 (HAG-100) 정도의 저분자 alginate를 제조하였으며, 이 저분자 alginate의 항돌연변이 효과와 cholesterol, glucose 및 카드뮴과의 결합능을 측정하여 그 물리${\cdot}$화학적 특성을 비교, 검토하였다. Ames test로 저분자 alginate의 항돌연변이 효과를 측정한 결과, $aflatoxin B_1 (AFB_1)$에 대한 간접돌연변이원성과 N-methyl-N'-nitro-N-nitrosoguanidine (MNNG)에 대한 직접돌연변이원성의 항돌연변이 효과는 HAG-10이 가장 높았고, 다음으로 HAG-50, HAG-100 및 alginate의 순으로 나타나 저분자화가 진행될수록 그 효과는 증가하는 것으로 나타났다. 저분자 alginate의 cholesterol, glucose 및 카드뮴에 대한 결합능은 alginate가 가장 높았고, 다음으로 HAG-50과 HAG-100은 유사하였으며, HAG-10은 결합능을 거의 보이지 않았다. To improve functionality and characteristics of alginate from the sea tangle, Laminaria japonicus, partially depolymerized alginates (HAG-10, average molecular weight 10,000; HAG-50, average molecular weight 50,000; HAG-100, average molecular weight 100,000) were obtained with hydrolysis of alginate by heating at $12^{\circ}C$. Effects of the depolymerization on physicochemical properties were investigated in the antimutagenicity and binding capacity of cholesterol, glucose and cadmium. In the Ames mutagenicity test using Salmonella typhimurium TA 100, HAG-10, HAG-50, HAG-100 and intact alginate reduced effectively the mutagenicities induced by aflatoxin $B_1 (AFB_1)$ and N-methyl-N'-nitro-N-nitrosoguanidine(MNNG) and HAG-10 showed the strongest antimutagenicity among the tested samples. The binding capacity of cholesterol, glucose and cadmium at different pH in vitro depended highly on molecular weight of alginate, and the changes in binding capacity at different pH was not different.

다시마 (Laminaria japonicus) Alginate의 가열가수분해에 따른 물리${\cdot}$화학적 및 생물학적 특성에 관한 연구 -4. $HCl{\cdot}ethanol$용액과 침수스트레스에 의해 유발되는 랫드 위궤양의 억제효과-

김육용,조영제,KIM Yuck-Yong,CHO Young-Je 한국수산과학회 2000 한국수산과학회지 Vol.33 No.6

Alginate를 가열에 의해 저분자화하여 저분자화에 따른 위점막 보호작용을 검토하기 위해 랫드에 있어서 저분자 alginate인 HAG-10, HAG-50, HAG-100 및 alginate의 투여 가 $HCl{\cdot}ethanol$ 용액과 침수스트레스에 의해 유발되는 위손상과 위궤양의 억제효과에 미치는 영향을 광학현미경으로 점막의 형태를 관찰하면서 그 억제율과 조직학적 변화를 측정하였다. $HCl{\cdot}ethanol$ 용액과 침수스트레스에 의해 유발되는 위손상과 위궤양에 대한 억제효과에 미치는 저분자 alginate의 영향을 검토한 결과, 그 억제효과는 alginate에서 가장 높았고, 다음으로 HAG-50과 HAG-100 및 HAG-10의 순으로 HAG-50, HAG-100 및 alginate에서 현저한 효과를 보였으나, HAG-10은 아무런 효과를 나타내지 않았다. 위손상과 위궤양점막을 광학현미경으로 관찰해 볼 때, 대조군과 HAG-10에서는 위점막 전체에 걸친 출혈과 부식에 의한 응고괴사가 진행되었고 점막 (粘膜)과 점막근판 (粘膜筋板) 및 근육층 (筋肉層)에 부종의 발생과 조직의 파괴가 일어났으나 HAG-50, HAG-100 및 alginate에서는 가벼운 병변이 진행되어 위점막층에 약간의 출혈만 관찰되어 위손상억제 및 위궤양 억제효과를 뚜렷히 나타내었다. Anti-ulcer effects of depolymerized alginate (HAG-10, average molecular weight 10,000; HAG-50, average molecular weight 50,000; HAG-100, average molecular weight 100,000) obtained by hydrolysis of alginate by heating at $121^{\circ}C$, against $HCl{\cdot}ethanol$ and water-immersion stress in rats were investigated. The acute gastritis, induced by $HCl{\cdot}ethanol$, and the gastric ulcer, induced by water-immersion stress, were inhibited dose-dependently by administration of HAG-50, HAG-100 and alginate. Histopathological lesions of the gastritis and gastric ulcer in rats treated with HAG-50, HAG-100 and alginate were significantly lower than those in rats fed with HAG-10. The inhibition rates (${\%}$) on acute gastritis induced by $HCl{\cdot}ethanol$ and gastritis ulcer induced by water-immersion stress in rats, were $13.00{\%}\;and\;15.74{\%}$of HAG-10, $41.15{\%}\;and\;35.72{\%}$ of HAG-50, $41.58{\%}\;and\;35.37{\%}$ of HAG-100, and $45.17{\%}\;and\;41.11{\%}$ of alginate, respectively. These results suggested that HAG-50, HAG-100 and slginate had a protective effect against the gastritis and gastric ulcer. The effect was not as visible when using HAG-10 in rats. From the present results, it was suggested that HAG-50 was an effective anti-ulcer agent against $HCl{\cdot}ethanol$and water-immersion stress in rats.

콜라겐을 알긴산으로 교차 결합시킨 인공진피의 창상치유 효과

장재훈,박대환,신정임,안기영,송철홍 대한성형외과학회 2006 Archives of Plastic Surgery Vol.33 No.1

Alginate, which is isolated from brown seaweed, is a bioabsorbable long chain polysaccharides, β-D-mannuronic acid and α-L-guluronic acid. The authors produced alginates and alginate-colllagen as a disc form. Then, to evaluate the efficacy of alginate and alginate-collagen complex as a wound healing material, three full-thickness skin defects of 2 cm in diameter were made at the back of the New Zealand white rabbits. Three groups of dressing materials-alginate, alginate-collagen complex and vaseline gauze as control group - were applied on the wound and the results were evaluated grossly and histopathologically. The authors compared gross findings of sizes of healed wound, wound epithelization and wound contraction by tracing the remaining wound area at 5th, 10th, 15th, 20th, 25th day after wound introduction, and wound biopsy was performed at 3rd, 7th, 14th, 21st day, respectively.Alginate and alginate-collagen complex showed statistically higher percentage of wound contraction and wound healing compared to control group(p<0.05). Alginate-collagen complex showed statistically higher percentage of wound contraction, epithelization and wound healing compared to alginate alone. In conclusion, the result suggests that alginate has a good effect of wound healing and that alginate-collagen complex is more effective in wound healing than alginate alone.

다시마 (Laminaria japonicus) Alginate의 가열가수분해에 따른 물리$\cdot$화학적 및 생물학적 특성에 관한 연구 5. 랫드의 체중, 장기, 췌장과 소장의 성분 및 소장융모의 미세구조에 미치는 저분자 Alginate의 영향

김육용,조영제,KIM Yuck-Yong,CHO Young-Je 한국수산과학회 2001 한국수산과학회지 Vol.34 No.1

Alginate를 가열에 의해 저분자화하여 저분자화에 따른 소화생리 특성을 검토하기 위해, 랫드에 저분자 alginate인 HAG-10,HAG-50, HAG-100 및 alginate를 장기간 섭취시켰을 때, 체중, 각종 장기의 무게와 길이 및 췌장과 소장의 소화효소활성과 단백질, DNA 및 RNA 함량을 측정하였고 소장응모의 미세구조의 변화를 관찰하였다. 랫드의 체중은 $5\%$와 $10\%$ HAG-50 및 $10\%$ alginate에서 체중의 증가가 유의적으로 현저히 억제되었다. 간장의 무게는 유의적인 차이가 없었고, 위의 무게와 길이는 $10\%$ HAG-10, $1\%$, $5\%$ 및$10\%$ HAG-50, $1\%$와 $5\%$ HAG-100 그리고 $5\%$ alginate에서 증가하는 경향을 보였으며, 췌장의 무게와 길이는 모두 약간 증가하였다. 소장과 맹장의 무게와 길이는 $1\%$와 $5\%$ HAG-10을 제외하고 모두 증가하였으나 대장의 무게와 길이는 전반적으로 감소하였다. 췌장의 amylase 활성은 유의적인 차이는 보이지 않았고, lipase 활성은 HAG-50에서 약간 저하하였으며, protease 활성은 $1\%$ HAG-10만 제외하고 유의적으로 저하하였다. 췌장의 단백질 함량은 모두 증가하였으나, DNA와 RNA 함량은 유의 적 인 차이가 없었다. 소장의 단백질 함량은 $5\%$와 $10\%$ HAG-50에서 그리고DNA는 $5\%$ HAG-50에서 가장 높은 함량을 보였으며, RNA는 전반적으로 증가하였다 소장융모의 미세구조는 HAG-50에서 주름이 많고 표면적이 넓은 잎사귀 모양의 응모세포와 돌림주름 및 Boblet cell이 현저히 발달되어 있었다. To examine functionality of depolymerized alginate obtained by hydrolysis of alginate through a heating process at $121^{\circ}C$ on gastrointestinal physiology, the changes of body weight, organ weight and length, pancreatic and small intestinal composition, and light microscopy (LM) observation of small intestinal microvilli's appearances were checked in the rats. Rats were fed diets containing $1\%, 5\%, and 10\%$ of each depolymerized alginate (HAG-10, HAG-50, HAG-100) and alginate for 35 days, The feeding of 5 and $10\%$ HAG-50 and $10\%$ alginate diets for 35 days significantly depressed the body weight gain, but increased the length and weight of the small intestine and cecum in rats (p<0.01). Pancreatic protease activity was decreased significantly (p<0.01) in all groups except lo/o of HAG-10 diets, but the protein content increased in all groups, However, pancreatic amylase and lipase activities as well as DNA and RNA content were not significantly different. The small intestinal protein and the DNA content were the highest in diets fed $5\%$ HAG-50; RNA content increased significantly (p<0.01) in all groups except in the fiber-free diets. Light microscopy (LM) observation showed growth of small intestinal microvilli with numerous ridges; the multiplication of the convolution goblet cells in rats fed with diets containing $5\%$ of HAG-50 were more than others group.

다시마 Alginate의 추출조건이 alginate 필름의 성질에 미치는 영향

유병진,심재만,장미화,You Byeong-Jin,SHIM Jae-Man,CHANG Mi-Hwa 한국수산과학회 1999 한국수산과학회지 Vol.32 No.5

생분해성이며 가식성 alginate 필름을 가공하기 위하여 추출용 액의 $Na_2CO_3$ 농도와 추출시간을 달리하여 alginate를 추출하고, 동 alginate로 제조한 alginate 필름의 수증기 투과율과 장력성의 변화를 측정하였다. 추출용액중의 $Na_2CO_3$ 농도의 증가 및 추출시간의 연장과 더불어 alginate의 점도와 중합도는 감소하였다. 저점도 alginate로 만든 alginate 필름은 높은 수증기투과율을 보였다. 가소제로서 사용된 sorbitol과 polyethylene glycol 첨가는 필름의 수증기투과율과 인장강도를 현저히 낮추는 효과를 보였다. Alginate의 추출시간은 alginate필름의 수중기투과율과 신장성에 미치는 영향이 적었으나 alginate 중합도는 인장강도에 직접적으로 영향을 미쳤다. The extracting conditions of alginates from sea tangle were evaluated by measuring water vapor permeability (WVP) and tensile properties of alginate film to obtain basic data of making an edible and biodegradable film. The alginates were extracted with $1\%,\;3\%$ and $5\%$ sodium carbonate ($Na_2CO_3$) for 1, 3, 5 and 10 hours, and the alginate film was made with various plasticizers. The higher concentration of $Na_2CO_3$ solution showed the lower viscosity and polymerization degree of alginate and the film prepared with alginates having low viscosity showed the higher WVP. The extracting hours had little effect on the WVP and the elongation of alginate film, but the polymerization degree of alginates directly affected the tensile strength of the film. The addition of sorbitol and polyethylene glycol as a plasticizer lowered the WVPs and the elongation of alginate film.

알지네이트 진피지지체 인공피부 개발: 예비보고

박대환,신정임 대한성형외과학회 2006 Archives of Plastic Surgery Vol.33 No.1

Alginate, a polymer of guluronic and mannuronic acid, is used as a scaffolding material in biomedical applications. The research was to produce highly-purified alginate from seaweeds and to evaluate the efficacy of alginate as dermal substrate. Our alginate purification method showed a production rate as high as 25%. The purified alginate contained little polyphenol contents and endotoxin, proteins. For study of wound healing, full thickness skin defects were made on the dorsal area of the animal models. And then alginate, fibroblast-growth- factor mixed alginate, alginate-collagen complex, vaseline gauze as control were applied on the wound, respectively, and were evaluated grossly and histopathologically. For biocompatibility test, alginate and alginate- collagen complex discs were implanted on the back of Sprague- Dawly rats. Four weeks after implantation, the animals were examined immunologically against alginate and collagen. Alginate and FGF-mixed alginate, alginate- collagen complex group showed statistically higher percentage of wound contraction and wound healing than control group(p<0.05). Alginate-collagen complex group and FGF-mixed alginate group showed statistically higher percentage of wound healing than alginate group. The experiment of biocompatibility and immunologic reaction against impanted alginate or collagen needs more investigation. Highly-purified alginate from seaweeds by our purification method, showed the effect of wound healing, and addition of FGF or collagen increases the alginate's wound healing effect. It shows the possibility of alginate as a dermal substrate.

다시마 alginate 점도 및 금속이온 결합능에 미치는 추출조건의 영향

유병진,임영선,정인학,You Byeong-Jin,IM Yeong-Sun,JEONG In-Hak 한국수산과학회 1998 한국수산과학회지 Vol.31 No.2

In order to investigate the physical properties of alginate extracted from sea tangle, Laminaria app., under various conditions, viscosity and binding capacity of metal ion (BCMI) of alginate were measured. The higher concentrations of sodium carbonate and the longer extracting time became, the lower apparent viscosity and BCMI were. BCMI in alginate reached maximum at the concentration of 0.06M metal ion. The BCMI of $Pb^{++}$ ion was the highest but $Cu^{++}$ ion was the lowest in the five metal ions. BCMI was increased in proportion as increase of viscosity in alginate. 추출조건에 따른 다시마 alginate의 성질을 조사하기 위해 추출용매인 $Na_{2}CO_{3}$의 농도를 $1\%,\;3\%$, 및 $5\%$로 조절하고, 추출시간을 1, 3, 5 및 10시간으로 하였을 때 각 추출조건에서 추출된 alginate의 점도변화와 금속이 온 결합능을 측정하였다. $Na_{2}CO_{3}$의 농도가 높을수록 추출시간이 길어질수록 alginate의 겉보기 점도 및 금속이온과의 결합능은 감소하였다. Alginate 의 최대 금속이온 결합능을 나타내는 이온농도는 0.06M, $25m{\ell}$였다. Alginate의 금속이온과의 결합능은 $Pb^{++}$이 가장 높았고, $Cu^{++}$이 가장 낮게 나타났다. Alginate의 점도가 증가할수록 alginate의 금속이온 결합능도 증가되었다.

대장균에서 분자 chaperone에 의한 alginate lyase의 가용성 발현 증대

남수완,신은정,이재형,박소림,김형락 한국생명과학회 2007 생명과학회지 Vol.17 No.2

When alginate lyase gene (aly) from Pseudoalteromonas elyakovii was expressed in E. coli, most of the gene product was produced as aggregated insoluble particles known as inclusion bodies. In order to produce a soluble and active form of alginate lyase, E. coli cells were cotransformed with the plasmids designed to permit coexpression of aly together with molecular chaperones such as DnaK/DnaJ/GrpE or GroEL/ES chaperones. The results revealed that the coexpression of aly together with DnaK/DnaJ/GrpE chaperone had a marked effect on the production of this protein as a soluble and active form, presumably through facilitating correct folding of alginate lyase protein. The optimal concentration of L-arabinose for the induction of DnaK/DnaJ/GrpE chaperone was found to be 0.05 mg/ml. When DnaK/DnaJ/GrpE chaperone was coexpressed, about 34% in the total alginate lyase was produced in the soluble fraction. By addition of 10% cetylpyridinium chloride, a clear zone around the colony coexpressing aly and DnaK/DnaJ/GrpE chaperone was formed, indicating that the alginate in the medium was hydrolyzed by active alginate lyase enzyme. E. coli에서 Pseudoalteromonas elyakovii 유래의 alginate lyase 유전자(aly)를 발현시킬 때, 대부분의 단백질이 불용성 내포체 형태로 발현됨을 확인하였다. Alginate lyase를 가용성 활성형으로 생산하기 위해 aly와 DnaK/DnaJ/GrpE 또는 aly와 GroEL/ES을 공발현하는 형질전환체를 얻었다. 공발현 결과, 단백질의 올바른 접힘을 도와주는 DnaK/DnaJ/GrpE chaperone이 가용성 및 활성형의 alginate lyase 생산에 매우 효과적임을 알 수 있었다. DnaK/DnaJ/GrpE chaperone의 발현에 유도제인 L-arabinose 최적 농도는 0.05 mg/ml이었으며, 이러한 공발현에 의해 약 34%의 alginate lyase가 가용성 분획에서 생산되었다. 또한 10%의 cetylpyridinium chloride를 첨가함으로써, 공발현 콜로니 주위에 투명환이 형성됨을 확인할 수 있었고, 이는 활성형 alginate lyase 효소에 의해 alginate가 분해되었음을 시사하였다.

Alginate/bacterial cellulose nanocomposite beads prepared using <i>Gluconacetobacter xylinus</i> and their application in lipase immobilization

Kim, Ji Hyun,Park, Saerom,Kim, Hyungsup,Kim, Hyung Joo,Yang, Yung-Hun,Kim, Yong Hwan,Jung, Sang-Kyu,Kan, Eunsung,Lee, Sang Hyun Elsevier 2017 Carbohydrate polymers Vol.157 No.-

<P><B>Abstract</B></P> <P>Alginate/bacterial cellulose nanocomposite beads, with well-controlled size and regular spherical shapes, were prepared in a simple manner by entrapping <I>Gluconacetobacter xylinus</I> in barium alginate hydrogel beads, followed by cultivation of the entrapped cells in culture media with a low sodium ion concentration. The entire surface of the alginate hydrogel beads containing the cells was covered with cellulose fibers (∼30nm) after 36h of cultivation. The cellulose crystallinity index of the alginate/bacterial cellulose beads was 0.7, which was slightly lower than that of bacterial cellulose prepared by cultivating dispersed cells. The water vapor sorption capacity of the alginate/bacterial cellulose beads increased significantly from 0.07 to 38.00 (g/g dry bead) as cultivation time increased. These results clearly indicate that alginate/bacterial cellulose beads have a much higher surface area, crystallinity, and water-holding capacity than alginate beads. The immobilization of lipase on the surface of the nanocomposite beads was also investigated as a potential application of this system. The activity and specific activity of lipase immobilized on alginate/bacterial cellulose beads were 2.6- and 3.8-fold higher, respectively, than that of lipase immobilized on cellulose beads. The alginate/bacterial cellulose nanocomposite beads prepared in this study have several potential applications in the biocatalytic, biomedical, and pharmaceutical fields because of their biocompatibility, biodegradability, high crystallinity, and large surface area.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Alginate/bacterial cellulose (BC) beads with well-controlled size and shape were prepared. </LI> <LI> Alginate/BC beads showed high surface area, high crystallinity, and high water-holding capacity. </LI> <LI> Alginate/BC beads were more favorable than alginate beads for lipase immobilization. </LI> <LI> Alginate/BC beads have many potential applications in biocatalytic, biomedical, and pharmaceutical fields. </LI> </UL> </P>

다시마 alginate와 bile acids의 결합능에 미치는 추출조건의 영향

유병진,임영선,정인학,이강호,YOU Byeong-Jin,IM Yeong-Sun,JEONG In-Hak,LEE Kang-Ho 한국수산과학회 1997 한국수산과학회지 Vol.30 No.1

추출조건에 따른 다시마 alginate의 성질을 조사하기 위하여 추출용매인 $Na_{2}CO_{3}$의 농도를 1, 3, 및 $5\%$로 각각 조절하고 추출시간을 각각 1, 3, 5 및 10시간으로 하였을때 각 추출조건에서 추출된 alginate의 bile acids 결합능과 물리화학적 특성을 조사한 결과는 다음과 같다. 건조다시마를 미세하게 마쇄할수록, $Na_{2}CO_{3}$ 농도와 추출시간이 증가할수록 alginate 추출량은 증가하였다. alginate의 bile acids와의 결합능은 저 농도의 $Na_{2}CO_{3}$ 용액으로 단시간에 추출한 것일수록 높게 나타났다. alginate에 의한 bile acids 결합능은 cholic acid에서 가장 높게 나타났고, taurocholic acid, glycocholic acid, deoxycholic acid 순으로 조사되었다. 다시마로 부터 alginate를 추출하고 남은 잔사(RAEA)의 bile acids와의 결합능은 alginate의 경우와 같이 추출 용액의 농도가 높을수록 추출시간이 길어질수록 감소하는 경향을 보였다. Bile acids와의 결합능은 cholic acid, taurocholic acid의 경우 alginate보다 다소 낮은 값을 나타냈고, glycocholic acid의 경우 alginate와 비슷한 값을 보였으나 deoxycholic acid와의 결합능은 추출된 alginate보다도 오히려 높은 값을 보였다. 추출된 alginate의 점도와 중합도는 추출용액의 농도가 증가하고 추출시간이 길어짐에 따라 감소하였으며 이에 비례하여 bile acid와의 결합능도 감소하였다. Changes in bile acid binding rapacity in vitro and physical properties of alginate extracted from sea tangle (Laminaria spp.) and residue after extracting alginate (RAEA) were investigated. For the purpose, extraction conditions controlled under 1, 3 and $5\%$ of sodium carbonate solution, and 1, 3, 5 and 10 hours of extraction time at $60^{\circ}C$. The less sea tangle had particle size and the higher concentration of sodium carbonate solution increseded, the more yield of alginate gained. High concentration of sodium carbonate solution and long extraction time resulted in weakly binding capacity in vitro by alginate. Among four bile acids, binding capacites with alginate were in the order of cholic, taurocholic acid>glycocholic acid>deoxycholic acid. The binding capacity of RAEA was rated at almost same degree of alginate. For increasing the binding capacity of bile acids by alginate, it was subject to high viscosity and degree of polymerization.