• Title/Summary/Keyword: ${\alpha}$-chitin

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The Esterification of Acetyltyrosine by $\alpha$-Chymotrypsin in EtOH/Water Mixture (에탄올 내에서 $\alpha$-Chymotrypsin에 의한 Acetyltyrosine의 에스테르화 반응)

  • 전유진;김세권
    • KSBB Journal
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    • v.9 no.3
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    • pp.312-318
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    • 1994
  • The esterification of Ac-Tyr-OH was carried out in one-phase system containing ethanol by ${\alpha}$-chymotrypsin. The results of the esterification reaction are as follows. Chitin-${\alpha}$-chymotrypsin complex was found to be an effective catalyst for the esterlfication of Ac-Tyr-OH in ethanol organic solvent. The optimal conditions for the esterification were chitn/${\alpha}$-chymotrypsin ratio, 20(w/w); reaction temp., $35^{\circ}C$; reaction pH, 8.0; reaction time, 24 hrs. Also, addition of chitin in water/water-miscible organic solvent was effective for the stability of the enzyme. The esterification yield, Km and Vmax under optimal conditions were 93%, 3.093mM and 1.088mM/mg/hr, respectively.

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Liquefaction and Saccharification of Starch Using $\alpha$-Amylase and Immobilized Glucoamylase ($\alpha$-아밀레이즈와 고정화된 글루코아밀레이즈를 이용한 전분의 액화 및 당화)

  • 안대희;장호남
    • Microbiology and Biotechnology Letters
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    • v.19 no.5
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    • pp.497-503
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    • 1991
  • The catalytic activities of immobilized gIucoamylase in a packed bed column and a continuous stirred tank reactor have been compared. Rapid production of glucose from liquefied starch have been studied through, the continuous liquefaction and saccharification using settling chamber. The immobilized glucoamylase with chitin gave the saccharification yield of 20% with the dextrin concentration of 100 g/l in a residence of 20 min. in a packed bed column. The half-life of immobilized glucoamylase with chitin was 19 days. The glucoamyalse immobilized in chitin and encapsulated with Ca-alginate gave the saccharification yield of 6% with the dextrin concentration of 50 g/l in a residence of 20 min. in a packed bed column. The Ca-alginate encapsulated and chitin immobilized glucomylase had a half-life of 25 days, which is 6 day larger than that of the immobilized glucoamylase with chitin only. In continuous liquefaction and saccharification, the glucose yield was 17% for the liquefied starch with naked barley concentration of 50gA in a residence of 20 min.

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Structural Characteristics of Insect Chitin/Chistosan (곤충유래 키틴/키토산의 구조특성)

  • Lee, Kwang-Gill;Lee, Yong-Woo;Kweon, Hae-Yong;Yeo, Joo-Hong;Park, In-Kyu;Nam, Jin;Seol, Kwang-Youl
    • Journal of Sericultural and Entomological Science
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    • v.40 no.2
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    • pp.158-162
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    • 1998
  • The chitin was isolated from various kinds of insects such as exuvia of Psacothea hilaris Pascoe, silkworm pupa, Agrius convolvuli or from cuticle of cockroach by treatment with dilute HCI and NaOH. The chemical and crystalline structure was characterized by FT-IR and X-ray diffractometer. All of the chitins extracted from insects showed characteristic ${\alpha}$-chitin peaks at the Bragg angle 2$\theta$=9.3$^{\circ}$, 19.4$^{\circ}$, and 23.5$^{\circ}$by X-ray diffraction analysis. The transition from chitin to chitosan was confirmed by IR spectra and the degree of deacetylation of the crab shell, silkworm pupa, cockroach, and Psacothea hilaris Pascoe was 70.9, 76.4, 75.5, and 74.1%, respectively. The double diffraction peaks of insect chitosan were observed at 2$\theta$=10$^{\circ}$and 20$^{\circ}$, indication the characteristic of hydrated crystalline structure of chitosan.

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Purification of Chitinase from an Antagonistic Bacterium Bacillus sp.7079 and Pro-Inflammatory Cytokine Gene Expression by PCTC

  • Han, Ok-Kyung;Lee, Eun-Tag;Lee, Young-Sun;Kim, Sang-Dal
    • Journal of Microbiology and Biotechnology
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    • v.13 no.1
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    • pp.77-84
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    • 2003
  • Chitinase was purified from an antagonistic bacterium Bacillus sp. 7079 by ammonium sulfate precipitation, QAE-Sephadex anion exchange chromatography, Sephadex G-100 gel filtration, and SP-Sephadex cation exchange chromatography. The molecula. weight of purified chitinase (PC-1) was approximately 66.5 kDa on SDS-PACE. PC-1 exhibited optimum pH and temperature of pH 7.5 and $45^{\circ}C$, respectively. More than $80\%$ of PC-1 was stable at pH 5.0 to 9.0, and more than $90\%$ at $40^{\circ}C$. $Fe^2+\;and\;Ca^2+$ inhibited the chitinase activity about $20\%$, and EDTA and p-CMB by about $30\%$, whereas $Ag^+$ inhibited the activity up to $65\%$. The $K_m$ value of PC-1 was 1.215 mg/ml with colloidal chitin as a substrate. We also investigated the effect of PC-1 treated chitin (PCTC) on the pro-inflammatory cytokine gene expression in macrophage RAW 264.7 cells. The expression of IL-$1{\alpha}$ and IL-$1{\beta}$ mRNA gene was investigated using reverse transcriptase polymerase chain reaction (RT-PCR). IL-$1{\alpha}$ and IL-$1{\beta}$ mRNA were induced by the treatment of PCTC and chitin only in RAW 264.7 cells. These expressions were induced as early as 2 h and sustained up to 24 h in RAW 264.7 cells. IL-$1{\alpha}$ and IL-$1{\beta}$ mRNA were more strongly expressed by the treatment of PCTC than chitin treatment alone in RAW 264.7 cells.

Chitin from Cuttlebone Activates Inflammatory Cells to Enhance the Cell Migration

  • Lim, Sung Cil;Lee, Ki-Man;Kang, Tae Jin
    • Biomolecules & Therapeutics
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    • v.23 no.4
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    • pp.333-338
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    • 2015
  • Our previous report showed that the extract from cuttlebone (CB) had wound healing effect in burned lesion of rat and the extract was identified as chitin by HPLS analysis. We herein investigated the morphology in CB extract using scanning electron microscope (SEM). Chitin was used as a control. There is no difference in morphology between CB extract and chitin. We also assessed the role of CB extract on the production of inflammatory mediators using murine macrophages and the migration of inflammatory cells. The extract induced the production of nitric oxide (NO) in macrophages. While the extract of CB itself stimulated macrophages to increase the expression of pro-inflammatory cytokines such as tumor necrosis factor (TNF)-${\alpha}$, interleukin (IL)-$1{\beta}$, and IL-6, CB extract suppressed the production of those cytokines by LPS. CB extract also induced the production of mouse IL-8 which is related to the cell migration, and treatment with CB enhanced fibroblast migration and invasion. Therefore, our results suggest that CB activates inflammatory cells to enhance the cell migration.

Chitin from the Extract of Cuttlebone Induces Acute Inflammation and Enhances MMP1 Expression

  • Lee, Ki Man;Shim, Hong;Lee, Geum Seon;Park, Il Ho;Lee, Ok Sang;Lim, Sung Cil;Kang, Tae Jin
    • Biomolecules & Therapeutics
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    • v.21 no.3
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    • pp.246-250
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    • 2013
  • We previously reported that the extract from cuttlebone (CB) has wound healing effect in burned lesion of rat. In present study, the main component of CB extract was analyzed and its wound healing activity was evaluated by using in vitro acute inflammation model. The extract of CB stimulated macrophages to increase the production of TNF-${\alpha}$. The extract also enhanced the production of TGF-${\beta}$ and VEGF, which were involved in angiogenesis and fibroblast activation. The treatment with CB extract enhanced proliferation of murine fibroblast. CB extract also induced the activation of fibroblast to increase the secretion of matrix metalloproteases 1 (MMP1). The constituent of CB extract which has wound healing activity was identified as chitin by HPLC analysis. The mechanism that the CB extract helps to promote healing of burned lesion is associated with that chitin in CB extracts stimulated wound skins to induce acute inflammation and to promoted cell proliferation and MMP expression in fibroblast. Our results suggest that CB or chitin can be a new candidate material for the treatment of skin wound such as ulcer and burn.

Effect of KGD1 Deletion on Cell Wall Biogenesis in Saccharomyces cerevisiae (Saccharomyces cerevisiae의 KGD1 유전자 결손이 세포벽 생합성에 미치는 영향)

  • Kim, Sung-Woo;Ahn, Ki-Woong;Park, Yun-Hee;Park, Hee-Moon
    • The Korean Journal of Mycology
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    • v.38 no.1
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    • pp.29-33
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    • 2010
  • KGD1 gene was cloned by functional complementation of defects in $\beta$-1,3-glucan synthase activity of the previously isolated Saccharomyces cerevisiae mutant LP0353, which displays a number of cell wall defects at restrictive temperature. We performed the gene disruption experiment to characterize the function of KGD1 gene, which encodes $\beta$-ketoglutarate dehydrogenase, in cell wall biosynthesis. The disruption of KGD1 showed the decreased growth rate, the increase of chitin synthases activity, alterations in cell wall composition, and increase of susceptibility to cell wall inhibitors such as Calcofluor white and Nikkomycin Z. These results suggested that KGD1 might be involved in cell wall biogenesis, especially the biosynthesis of $\beta$-1,6-glucan and chitin in S. cerevisaie.

Characterization of Cellulase from Bacillus subtilis NSC Isolated from Soil (토양으로부터 단리한 Bacillus subtilis NSC 유래 Cellulase의 특성 규명)

  • Kim, Sang Jin;Park, Chang-Su
    • Journal of Chitin and Chitosan
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    • v.23 no.4
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    • pp.228-233
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    • 2018
  • We isolated microorganisms from soil, which is sampled at forest, Kyeonbuk, Korea, as cellulolytic microorganisms. The isolated strains were identified by analysis of 16S rRNA gene from the starins. The result, four kinds of Bacillus subtilis, one kind of Bacillus amyloliquefaciens, and one kind of Bacillus cereus were identified. Among these strains, Bacillus subtilis was selected due to its high cellulase activity and this strain was named as Bacillus subtilis CNS. The optimum pH and temperature of the cellulase from Bacillus subtilis CNS was pH 5.0 and $40^{\circ}C$, respectively. In the investigation of pH and temperature stability, the cellulase from Bacillus subtilis NSC stabled pH 4.0~6.0 range and until $40^{\circ}C$ for 30 min perfectly. In the enzyme activity for various cellulosic substrate, cellulase from Bacillus subtilis CNS showed the highest activity for CM-cellulose. And, the enzyme activities for alkali swollen cellulose, Alpha-cellulose, Sigmacell-cellulose, and Avicel were approximately 31%, 8%, 8% and 4% of activity for CM-cellulose, respectively. In the degradation of CM-cellulose, the 0.26 U/ml and 0.52 U/ml of cellulase showed 0.43 and 0.76 U/ml activity for CM-cellulose after the reaction of 120 min, respectively.

Study of the organic and mineral composition of living pupae of the wild silkworm Saturnia pyri for use as food additives

  • Shukurova, Zarintac Yusif;Khalilov, Zarbali Murad;Shukurlu, Yusif Hacibala
    • International Journal of Industrial Entomology and Biomaterials
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    • v.43 no.2
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    • pp.52-58
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    • 2021
  • The article presents the results of the content of the chemical and biochemical composition of the pupa of the wild silkworm Saturnia pyri belonging to the family Saturniidae, species of Lepidoptera. The nutritional value of silkworm Saturnia pyri pupae was evaluated, which contained 51% dry matter, 52.50% crude protein, 27.89% fat, 10.50% chitin fibers, 2.5% ash and 27 macro- and microelements and 25 mg alpha tocopherols in 100 g oil. The X-ray fluorescence method was used to determine the content of mineral elements in the pupa of the silkworm Saturnia pyri. It was revealed that the pupa of this type of silkworm contains 25 elements, of which the relative amount of K, Mg, Na, Ca, Al is much higher than other elements.

Anti-allergic Effect of Eckolona cava Ethyl Acetate Fraction of on IgE/BSA-stimulated Bone Marrow-derived Cultured Mast Cells (IgE/BSA가 자극한 골수유래 비만 세포에 대한 감태 Ethyl Acetate 분획물의 항알러지 효능)

  • Han, Eui Jeong;Kim, Hyun Soo;Shin, Eun Ji;Kim, Min Ju;Han, Hee-Jin;Jeon, You-Jin;Jee, Youngheun;Ahn, Ginnae
    • Journal of Chitin and Chitosan
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    • v.23 no.4
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    • pp.277-284
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    • 2018
  • In this study, we investigated the anti-allergic effect of the ethyl acetate fraction of Ecklonia cava (EC-EtoAc) on the immunoglobulin E (IgE)/bovine serum albumin (BSA)-mediated activation of bone marrow-derived cultured mast cells (BMCMCs). We revealed that the $62.5{\mu}g/ml$ of EC-fractions ($EC-CHCl_3$, EC-Hexane and EC-EtoAc) inhibited IgE/BSA-activated ${\beta}$-hexosaminidase release from BMCMCs without cytotoxicity. Especially, EC-EtoAc showed the higher ${\beta}$-hexosaminidase release than the others. Also, EC-EtoAc reduced the expression levels of cytokines such as interleukin (IL)-$1{\beta}$, IL-4, IL-5, IL-6, IL-10, IL-13, interferon (IFN)-${\gamma}$ and tumor necrosis factor (TNF)-${\alpha}$ and a chemokine, thymus- and activation-regulated chemokine (TARC), compared to the only IgE/BSA-treated BMCMCs. Furthermore, EC-EtoAc significantly prevented the binding of IgE to Fc epsilon receptor $(Fc{\varepsilon}R)I$ and reduced the $Fc{\varepsilon}RI$ expression on the sensitized BMCMCs. Taken together, these results suggest that E. cava may be the natural agent with beneficial potentials for the treatment of type I allergic diseases induced by mast cell activation.