• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.34 no.5
• /
• pp.563-569
• /
• 2001

In order to utilize the processing wastes of squid, chitosan was prepared by intermittent deacetylation treaoent of $\beta-chitin$ contained richly in the pen of squid. Acetylchitosan also was synthesized from squid pen chitosan with anhydrous acetic acid and their characteristics were investigated. The amounts of nitrogen and ash of squid pen chitosan were $5.80.2\% and 0.2\pm0.03\%$ respectively, the yield of squid pen chitosan was $25\pm3\%$, the degree of deacetylation was $92\%$, and the molecular weight was $1.15\times10^6$, Acetyl contents of N-acetylchitosan powder, acetylchitosan bead, N-ACF-1 (N-acetylchitosan film-1) and N-ACF-2 (N-acetylchitosan film-2) were $55.9\%, 63.2\%, 56\% and 58.7\%$ respectively. Two major peaks, amide I ($1,653 cm^{-1}$) and II ($1,558 cm^{-1}$) bent, on FT-IR spectra of the N-acetylchitosan from squid pen were almost similar to these of $\beta-chitin$, While there was a broad single peak at $1,601 cm^{-1}$assigned to be an amide I bend in squid pen chitosan. The CP/MAS NMR spectra of $\beta-chitin$, squid pen chitosan and N-acetylchitosan from squid pen showed a relative broad and single peak at 74 ppm assigned to fifth carbon (C-5) and third carbon (C-3). In case of $\beta-chitin$ and N-acetylchitosan from squid pen, single peak at 74 ppm was showed as the same of $\beta-chitin$ type.

• Journal of Ginseng Research
• /
• v.34 no.2
• /
• pp.98-103
• /
• 2010

The medicinal plant Panax ginseng (P. ginseng) contains various phytosterols and bioactive triterpene saponins (ginsenosides). Squalene synthase catalyzes the first committed step in ginsenoside biosynthesis. Transgenic plants of P. ginseng were generated by introducing the squalene synthase gene derived from P. ginseng. Adventitious roots of the transgenic ginseng grew best in B5 medium, and 2 g of inoculum secured an optimal growth rate. Two phytohormones, indolebutyric acid and 1-naphtalene acetic acid, increased root growth and decreased ginsenoside production. Treatment with two selected elicitors, chitosan and jasmonic acid, and a precursor of the isoprenoid pathway, mevalonic acid, enhanced ginsenoside production and retarded ginseng growth rate.

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.25 no.1
• /
• pp.51-57
• /
• 1992

The preservative effect of chitosan film packing on quality of lightly-salted and dried horse mackerel was studied. In preparation of chitosan film, blue crab shell chitosan was dissolved in dilute acetic acid$(1.0\%,\;v/v)$, filtered, and spreaded on plastic plate and dried at $50\pm2^{\circ}C$. The chitosan film thus obtained was neutralized with 1.0N NaOH for 2 hrs and dried at room temperature after washing several times with distilled water. The lightly-salted and dried horse mackerel product was prepared by drying for 4 hrs at $40\pm2^{\circ}C$ in hot air dryer after packing with the chitosan film. During storage at $5.0\pm0.5^{\circ}C$, moisture content of the product was higher than that of the reference, but contents of VBN(volatile basic nitrogen) , amino nitrogen, and TMA of the product on dry basis were lower than those of the reference. Viable cell count, TBA value, and peroxide value of the product were also lower than those of the reference. Judging from the result of sensory evaluation, the chitosan film packing in the storage of lightly-salted and dried horse mackerel was remarkably elongated shelf-life of the product. From the results of chemical and sensory evaluation, it was concluded that chitosan film packing was an effective method for retaining the quality of lightly-salted and dried horse mackerel.

• Macromolecular Research
• /
• v.17 no.10
• /
• pp.734-738
• /
• 2009

Biodegradable, pH-sensitive, glycol chitosan (GC) hydrogels were prepared using divinyl adipate (DVA) as a crosslinker and acetic acid as a catalyst. DVA has highly reactive double vinyl ester groups and GC contains a high density of hydroxyl groups, with two in every glucosamine unit. The transesterification reaction between vinyl esters and hydroxyl groups produced crosslinked GC hydrogels. The initial crosslinking reaction was monitored by measuring the viscosity of the reaction mixture. When DVA was added to the GC solution and heated to $50^{\circ}C$, the viscosity of the GC solution gradually increased, implying a crosslinking reaction and hydrogel formation. A new peak from the ester group was observed in the FTIR spectra of the GC hydrogels, confirming the crosslinking reaction. The synthesized GC hydrogel showed pH-dependent water absorbency, mainly due to the presence of amine groups ($-NH_2$) at the C-2 position of the glucosamine unit of GC. The water absorbency greatly increased at acidic pH and slightly decreased at alkaline pH. The GC hydrogel gradually degraded in $37^{\circ}C$ water due to hydrolysis of the ester bonds, which were intermolecular crosslinking sites. A red dye, 5-carboxyltetramethyl-rhodamine (CTMR), was entrapped in the GC hydrogels as a model compound. CTMR was released from GC hydrogels in two steps: an initial burst release mainly due to desorption and diffusion, and a second sustained release possibly due to gradual degradation.

• Biotechnology and Bioprocess Engineering:BBE
• /
• v.5 no.5
• /
• pp.345-349
• /
• 2000

To easily separate chitosanoligosaccharides by size exclusion, an coencapsulating technology of substrate and enzyme was developed. The membrane was composed of alginate and a divalent cation such as calcium. Chitosan and chitosanase were enveloped in this membrane and the product released to medium by size exclusion. The capsule was stabilized in a 2% acetic acid solution (pH 5.0) containing 0.145 M CaCO$_3$. The leakage of substrate caused by the agitation speed was controlled by increasing alginate and CaCO$_3$concentrations. The lower limit of the alginate concentration and the agitation speed were 0.5% and 49rpm, respectively. Membrane thickness and capsule diameter were 10$\mu\textrm{m}$ and 2.5mm, respectively. By TLC analysis, the composition of chitosanoligosaccharides were mainly 3-6 mers. The molecular weight distribution of the released oligosaccharides ranged from 262 to 3624 Da by GPC.

• Proceedings of the Membrane Society of Korea Conference
• /
• 2003.07a
• /
• pp.61-64
• /
• 2003

Optical resolution of a-amino acid (tryptophan and tyrosine) optical isomers was achieved by a pressure driven membrane separation process, using self-supporting crosslinked membranes base on polysaccharide with different swelling indices that ranged from 100 to 70%. The membranes prepared by casting and drying the polymer solution containing 5wt% acetic acid on an acryl plate followed by crosslinking with glutaraldehyde were characterized using such analytical methods as FTIR and swelling index measurements. On the way of separating the optical isomers, several experimental factors such as the concentration of the feed solutions, operating pressure and temperature, and degree of crosslinking of the membranes have been studied. When the chitosan membranes with 70% of swelling index were used , almost complete optical resolution was obtained; 97.92% of enantiomeric excess (ee %) and 2.26 g/$m^2$ㆍh of flux. The operating pressure and the concentration of feed solutions were respectively 1.0 kgf/$\textrm{cm}^2$ and 0.49 mmol/L.

• Journal of Microbiology and Biotechnology
• /
• v.15 no.3
• /
• pp.626-632
• /
• 2005

A new bacterium BL-2 excreting a novel cationic polyglucosamine biopolymer was isolated from the spoiled leaves of Chinese cabbage and identified as Enterobacter sp. BL-2. The isolated Enterobacter sp. BL-2 was cultivated in pH-stat fed-batch culture using acetic acid as the feeding stock at pH 8.0, resulting in 17.11 g/l of cells and 1.53 g/l of an extracellular biopolymer after 72 h. The excreted biopolymer was purified by a three-step procedure, involving ethanol precipitation and deproteinizations, to a nearly homogeneous state, and its molecular weight was found to be 106 kDa. It was composed of glucosamine, rhamnose, and galactose at a molar ratio of 86.4:1.6:1.0, respectively, indicating a rarely found novel high-glucosamine-containing biopolymer. The FT-IR and $^{13}C-NMR$ spectra of the novel cationic polyglucosamine biopolymer PGB-l revealed a close identity with chitosan from crab shell. It can effectively flocculate various suspended solids, including kaolin clay, $Ca(OH)_2,\;Al_{2}O_3$, active carbon, microbial cells, and acidic dyes.

• Polymer(Korea)
• /
• v.36 no.3
• /
• pp.380-392
• /
• 2012

Four kinds of N,O-hydroxypropyl chitosans (HPCTOs) with degree of substitution(DS) and molar substitution (MS) ranging from 2.47 to 2.52 and 4.9 to 7.8, respectively were synthesized, and their molecular chracteristics and thermotropic and lyotropic liquid crystalline properties were investigated. MS was exceedingly larger than DS, showing that in the later stages of reaction, propylene oxide was preferentially added to the side chains rather than the main chain. All the derivatives formed thermotropic cholesteric phases. The glass and clearing temperatures were decreased with increasing MS. The optical pitches (${\lambda}_m$'s) of the thermotropic cholesteric phases increased with temperature. However, the ${\lambda}_m$'s of the derivatives at the same temperature increased with increasing MS. Solutions of HPCTOs in water, methanol, ethanol, acetic acid, and formic acid containing more than 30 wt% polymer also formed cholesteic phases whose ${\lambda}_m$'s decreased exponentially with increasing polymer concentration. The concentration dependence of ${\lambda}_m$ of HPCTO solutions, however, highly depended on the nature of the solvent and MS. The thermotropic and lyotropic mesophase properties of HPCTOs were significantly different from those reported for hydroxypropyl celluloses. The results indicate that the secondary amino group in the C-2 position plays an important role on the formation, stabilization, and temperature and concentration dependencies of ${\lambda}_m$ of the cholesteric mesophase.

• KSBB Journal
• /
• v.16 no.4
• /
• pp.321-326
• /
• 2001

The change of molecular weight inside and outside a capsule produced using coencapsulating technology was investigated. Chitosan and chitosanase were enveloped in this membrane and product released was a loaded the medium by the principle of size exclusion. The leakage of substrate corresponding to the agitation speed was controlled by adjusting the alginate and CaCO$_3$ concentrations. The optimal condition of alginate concentration and agitation speed were 0.5% and 40rpm, respectively. Membrane thickness and capsules diameter were 10 $\mu$m and approx. 3.0 - 1.5 mm, respectively. Molecular weight difference by concentration and alginate viscosity were of little significance. In accordance with the molecular weight distribution versus enzyme concentration relationship, low concentration of enzyme produced high molecular weight oligosaccharides. At a 1.5 mm capsule size the product diffusion rate to outer surface highest. The molecular weight distribution of the released oligosaccharides was ranged from 1000 to 6000 Da. More than 80% of the initial activity of encapsulated enzyme retained after 8hrs of reaction.

• Journal of Periodontal and Implant Science
• /
• v.33 no.1
• /
• pp.27-35
• /
• 2003

1. 목적 생체재료의 생체친화성을 증진시키고 치유를 촉진하기 위한 목적으로 생체재료의 생화학적 표면개질에 관한 연구가 널리 진행되고 있다. 이와 같은 목적으로 이용되어 온 부착분자에는 아미노산, 펩타이드, 단백질, 효소 및 성장인자들을 들 수 있으며, 이들 분자들을 금속, 골대체물질 및 폴리머와 같은 생체재료의 표면개질에 이용하여 왔다. 이 연구의 목적은 생체적합성이 우수하고 생분해성을 지닌 키토산으로 얇은 막을 제작한 후, 세포외 기질의 구성성분 중 세포부착에 관여하는 RGD 펩타이드를 부착시킨, 표면개질 키토산막의 생물학적 영향을 MG-63 조골양세포를 이용하여 관찰하는 것이다. 2. 방법 2% acetic acid에 키토산 가루를 녹여 만든 2% 키토산 용액으로 24-well 배양접시의 표면을 도포 후 24시간 동안 건조시켜 키토산막을 제작하였다. GRGDS 펩타이드를 cross-linker(EDC, NHS) (Sigma, MO, USA) 용액과 반응시켜서 펩타이드의 카르복실기를 활성화시켰다. 이들을 PBS 완충용액으로 수화시킨 키토산막과 결합시켜 펩타이드의 활성화된 카르복실기와 키토산의 아민기 간에 안정적인 아미드 결합(amide bond)이 형성되도록 하였다. 하루 동안 반응을 일으킨 후 PBS 완충용액과 증류수로 씻어내고 냉동 건조시킴으로써 GRGDS가 결합된 키토산막을 제작하였다. 재료 표면의 화학 성분을 알아보는데 사용되는 방법의 일종인 X-ray photoelectron spectroscopy(XPS) 분석을 통하여 부착분자가 키토산막에 결합된 여부를 확인하였다. GRGDS 펩타이드에 요오드를 결합시킨 후, 이것을 키토산막에 공유 결합시키고 XPS를 통해 요오드가 재료 표면에서 검출되는지를 검사하였다. 요오드가 검출된다면 이것은 키토산막 표면에 실제로 GRGDS 펩타이드가 존재하는 것을 의미하게 된다. 표면개질된 키토산막에 사람조골양세포인 MG-63을 접종하여 이를 실험군으로 하였고, 표면이 개질 되지 않은 키토산막을 대조군으로 하였다. 세포부착의 최적화 농도를 확인하기 위하여 GRGDS를 0.01, 0.05, 0.1, 0.25, 0.5, 1.0mg/ml의 농도로 준비하였다. 배양 후 1일, 7일째에 각 well에서 trypsin EDTA를 이용하여 세포를 분리한 후, 이를 원심 분리하여 세포수측정기를 이용하여 부착 세포의 수를 측정하여 세포의 부착 정도를 비교하였다. 배양 2시간, 24시간 후 주사전자현미경을 이용하여 키토산막에 부착된 세포의 양상을 관찰하였다. 3. 결과 XPS를 통한 표면의 화학 성분 분석 결과 GRGDS 펩타이드를 결합시킨 키토산막에서 요오드가 검출되었으며 펩타이드를 부착하지 않은 대조군에서는 검출되지 않았다. 따라서 cross-linker를 이용한 펩타이드와 키토산막의 공유결합을 확인할 수 있었다. 세포 배양 후 1일째 부착된 세포 수를 측정한 결과 0.1mg/ml 이상의 GRGDS 펩타이드 농도로 공유 결합시킨 키토산막에서 부착 세포 수가 다른 농도에 비해 유의성 있게 많이 관찰되었다. 이 농도 이하에서는 대조군과 실험군간에 세포부착의 유의한 차이가 없었다. 따라서 주사전자현미경을 이용한 부착 세포의 양상에 관한 관찰은 0.1mg/ml 농도의 펩타이드를 이용하였다. 세포 배양 7일째, 부착된 세포 수 측정 결과 GRGDS의 농도에 따른 유의성 있는 차이가 없었으며, 실험군과 대조군간에도 유의성 있는 차이가 없었다. 주사전자현미경 관찰결과 2시간 및 24시간 배양된 실험군 모두에서 별모양의 세포들이 키토산막 표면에 편평하게 잘 부착되어 있으며 많은 위족이 발달된 소견을 보인 반면, 대조군에서는 원형 또는 다각형 모양의 세포들이 실험군에 비해 부착이 덜 되어있는 양상을 보였다. 이 연구를 통하여 기능성 펩타이드를 생체재료의 표면에 공유결합 시키는 방법을 확립할 수 있었으며, RGD 펩타이드의 공유결합으로 표면개질된 키토산막이 조골세포의 부착능을 증진시킬 수 있음을 확인하였다. 표면개질된 생체재료를 소, 중동물에 적용시켜 생체 내에서의 생물학적 영향을 평가할 필요가 있으며, 이 실험의 결과는 향후 다양한 기능성 부착분자를 선발, 고안하여 임플란트용 생체재료의 표면개질에 이용하는 이른바 모방생체재료분야에 널리 활용될 수 있을 것으로 생각된다.