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The Korean Society for Biotechnology and Bioengineering (한국생물공학회)
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Extraction of Glycosaminoglycans from Styela clava Tunic

미더덕 껍질로부터 Glycosaminoglycans의 추출

  • 안삼환 (경상대학교 해양생물이용학부, 해양산업연구소) ;
  • 정성훈 (경상대학교 해양생물이용학부, 해양산업연구소) ;
  • 강석중 (경상대학교 해양생물이용학부, 해양산업연구소) ;
  • 정태성 (경상대학교 수의학과) ;
  • 최병대 (경상대학교 해양생물이용학부, 해양산업연구소)
  • Published : 2003.06.01
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Abstract

Glycosaminoglycans (GAGS was extracted from sea squirt, Styela clava with sodium phosphate at 105$^{\circ}C$ for 2 hr and deproteinized with trichloroacetic acid or hydrochloride. The GAGs obtained from tunic consist 41.7% crude carbohydrates, 31.8% crude protein, and 31.2% sulfate. It was mainly constituted of galactose, glucosamine, glucose, mannose, and glacrosamine. The prominent amino acid were phenylalanine, threonine, glutamic acid, and aspartic acid. Mineral contents was mainly constituted 3.0 mg% sodium, 1.6 mg% potassium, and 1.2 mg% phosphorus. Trichloroacetic acid, hydrochloride and 5-sulfosalicylic acid were used for deprotein of the GAGs. Effective volume for deprotein of crude GAGs were 5.0% trichloroacetic acid (w/v) and 10.0% HCI (v/v) treatment. The deproteinized GAGs contained 35.1%, 35.4% of protein and 22.0%, 18.5% of sulfate, respectively.

GAGs의 추출 방법 중 효소의 사용으로 추출 효율의 상승과 원료수급의 문제점을 해결할 수 있었다. 그러나 효소 사용에 따른 생산단가의 상승과 단백질 함량의 증가에 따른 화장품 규격을 만족시킬 수 없음으로, 1/60 M sodium phosphate buffer로 105$^{\circ}C$로 열수 추출하는 것이 화장품 규격을 만족시키는 방법으로 나타났다. 이렇게 추출한 GAGs의 SO$_4$ 함량은 31.2%이고 회분의 함량이 22.2%로 GAGs 추출 향상을 위해 사용되어지는 sodium phosphate의 사용으로 회분 함량이 높아졌음을 알 수 있고 무기질 분석에서 Na의 함량이 3.0 mg%로 총 무기질의 47.6%를 차지하고 있었다. 시료를 HPLC로 분석하였을 때 chondroitin sulfate A, C와 동일한 분석시간을 나타내어 chondroitin sulfate의 함유 가능성을 나타내고, glucosamine과 galactose가 78.0% 존재하는 것을 당조성 분석을 통해 확인하였다. 일반 성분, HPLC분석, 당 조성분석과 아미노산 분석결과로 glucosamine과 galacturonic acid가 주가 되어 threonine으로 연결된 구조를 하는 GAGs라는 것을 알 수 있다. 화장품 원료 기준에 맞추기 위해서는 sulfate 함량이 35.0~45.0%, 단백질 함량은 14.0~22.0%로 유지해야하므로 제단백결과 5.0%, 10.0%, 20.0% TCA (w/v) 처리, 10.0%, 20.0%, 40.0% HCI (v/v) 처리, UF(ultra filtration)를 포함한 10.0% TCA (w/v), 20.0% SSA (w/v), 25.0% HCI (v/v) 처리 결과 5.0% TCA (w/v) 및 10.0% HCI (v/v)이 화장품 기준에 적합하며 경제적이며 효율적이라는 것을 알 수 있었다.

Keywords

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