Comparative Study of Digestive Enzymes of Squid (Todarodes pacificus) Viscera after Supercritical Carbon Dioxide and Organic Solvent Extraction

Md. Salim Uddin,Hyang-Min Ahn,Hideki Kishimura,전병수 한국생물공학회 2009 Biotechnology and Bioprocess Engineering Vol.14 No.3

Three major classes of digestive enzymes of squid viscera were characterized following extraction of oil by supercritical carbon dioxide (SCO2) and organic solvent, n-hexane. Squid viscera were extracted at temperature, 35~45°C and pressure, 15~25 MPa for 2.5 h by SCO2 with a constant flow rate of 22 g/min. Oil extraction yield increased with the increasing of extraction pressure and temperature. The highest oil extracted residues of squid viscera were used for characterization of digestive enzymes. The activities of protease, lipase, and amylase were highest in n-hexane treated squid viscera samples and lowest in SCO2 treated samples. The crude extracts of SCO2 and n-hexane treated squid viscera samples showed almost same optimum pH and pH stability for each of the digestive enzymes. The optimum temperature of protease, lipase, and amylase were found to almost similar in SCO2 and n-hexane treated samples. But the thermal stability for each digestive enzyme in SCO2 treated squid viscera were slightly higher than that of n-hexane treated squid viscera. Studies using SDS-PAGE showed no significant differences in protein patterns of the crude extracts of untreated and SCO2 and n-hexane treated squid viscera indicating no denaturation of proteins

오징어 내장의 초임계 이산화탄소 추출 잔류물로부터 인지질의 분리

유병욱 ( Pyoung Ook U ),전병수 ( Byung Soo Chun ) 한국화학공학회 2010 Korean Chemical Engineering Research(HWAHAK KONGHA Vol.48 No.6

Phospholipids were recovered from squid viscera residues by ethanol extraction after supercritical carbon dioxide(SCO2) extraction and from squid viscera was not processed SCO2 by various organic solvent extraction. SCO2 extraction were performed at 45˚C and 20MPa for removal of non polar lipid molecules from freeze dried squid viscera sample. Phospholipids were extracted from freeze dried squid viscera sample by chloroform, hexane, methanol, and ethanol and from SCO2 extracted squid viscera sample by ethanol. The pH was fixed at 5.7 for all phospholipids extraction conditions. Phospholipid classes were analyzed by HPLC equipped with evaporative light scattering detector (ELSD). Phosphatidyl choline (PC) extracted by ethanol from SCO2 extracted residues was higher than that of extracted by ethanol from squid viscera. But phosphatidyl ethanolamine (PE) and phosphatidic acid (PA) were extracted higher percentage in raw squid viscera. The fatty acid compositions in phospholipids extract by ethanol extract from SCO2 extracted residues were analyzed by gas chromatography (GC). Docosahexanoic acid (DHA) was found in highest percentage in phospholipid extract.

Fatty Acids and Protein Recovery of Squid Viscera with Supercritical Carbon Dioxide

Park, Ji-Yeon,Back, Sung-Sin,Chun, Byung-Soo The Korean Society for Marine Biotechnology 2006 한국해양바이오학회지 Vol.1 No.3

Supercritical carbon dioxide ($SCO_2$) extraction was investigated as a method for protein-sourcing material from squid viscera. To find the optimum conditions, the extraction of squid viscera using $SCO_2$ was performed under the conditions of temperature range from 35 to $45^{\circ}C$ and constant pressure 25 MPa using Hewlett-Packard 7680T. Also from result of SDS-PAGE, the protein denaturation was minimized when using $SCO_2$ extraction. And the major amino acids in the squid viscera were glutamic acid, aspartic acid, lysine, leucine, arginine, alanine, glycine, isoleucine, and valine. The main fatty acids from squid viscera were myristic acid, palmitic acid, stearic acid, heneicosanoic acid, palmitoleic acid, elaidic acid, oleic acid, eicosenoic acid, EPA (eicosapentaenoic acid), and DHA (docosahexaenoic acid).

The Storage Property of Squid Viscera by Supercritical Carbon Dioxide Extraction

Lee, Min-Kyung,Yoo, Hong-Suk,Pack, Hyun-Duk,Chun, Byung-Soo The Korean Society for Marine Biotechnology 2007 한국해양바이오학회지 Vol.2 No.1

The oil and concentrated protein powder from squid viscera was extracted and recovered by a semi-batch supercritical carbon dioxide ($SCO_2$) extraction system and the degree of oxidation in the extracted oil was measured in order to compare with extracted oils using organic solvents. The degree of storage in treated squid viscera by $SCO_2$ extraction was measured in order to compare with untreated squid viscera. As results obtained, it was found that the auto-oxidation of the oils using $SCO_2$ extraction occurred very slowly compared to the oils by organic solvent extraction. And the treated squid viscera by $SCO_2$ extraction was reached the point of initial rottenness slowly than untreated squid viscera.

Stability of Antioxidant Properties and Essential Amino Acids in Squid Viscera Hydrolysate Produced using Subcritical Water

김령희,A. K. M. Asaduzzaman,유치홍,전병수 한국수산과학회 2013 Fisheries and Aquatic Sciences Vol.16 No.2

Subcritical water hydrolysis was carried out in a batch reactor to produce valuable materials, such as low-molecular weight (MW) peptides and essential amino acids with antioxidant properties, from heat-dried squid viscera. Hydrolysis of squid viscera was performed at 160 to 280°C for 3 min. The yield was increased by increasing the temperature and pressure, while the protein content of squid viscera hydrolysate decreased with increasing temperature. Low-MW peptides were detected in all hydrolysates by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The highest yields of free and structural amino acids in heat-dried squid viscera hydrolysate were at 160°C and were 411.95 ± 1.15 and 346.62 ± 1.25 mg/100 g, respectively. All essential amino acids were detected in viscera hydrolysates; leucine was the most abundant. Antioxidant activities of hydrolysates were highest at 220°C. Greater than 98 ± 0.26% of the ABTS antioxidant activity was retained in hydrolysates after long-term heat treatment.

Stability of Antioxidant Properties and Essential Amino Acids in Squid Viscera Hydrolysate Produced using Subcritical Water

Kim, Ryoung-Hee,Asaduzzaman, A.K.M.,You, Chi-Hong,Chun, Byung-Soo The Korean Society of Fisheries and Aquatic Scienc 2013 Fisheries and Aquatic Sciences Vol.16 No.2

Subcritical water hydrolysis was carried out in a batch reactor to produce valuable materials, such as low-molecular weight (MW) peptides and essential amino acids with antioxidant properties, from heat-dried squid viscera. Hydrolysis of squid viscera was performed at 160 to $280^{\circ}C$ for 3 min. The yield was increased by increasing the temperature and pressure, while the protein content of squid viscera hydrolysate decreased with increasing temperature. Low-MW peptides were detected in all hydrolysates by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The highest yields of free and structural amino acids in heat-dried squid viscera hydrolysate were at $160^{\circ}C$ and were $411.95{\pm}1.15$ and $346.62{\pm}1.25$ mg/100 g, respectively. All essential amino acids were detected in viscera hydrolysates; leucine was the most abundant. Antioxidant activities of hydrolysates were highest at $220^{\circ}C$. Greater than $98{\pm}0.26%$ of the ABTS antioxidant activity was retained in hydrolysates after long-term heat treatment.

Koji를 첨가하여 발효한 오징어 내장 조미료의 품질특성

최승화(Seung Hwa Choi),김상무(Sang Moo Kim) 한국식품영양과학회 2011 한국식품영양과학회지 Vol.40 No.1

본 연구는 오징어 가공부산물인 내장에 koji를 첨가하여 속성 발효한 천연조미료 소재 개발을 목적으로 결과를 요약하면 다음과 같다. 휘발성 염기질소량과 trimethylamine은 대조군이 가장 급격하게 증가하였고, 식염이 많이 첨가될수록 서서히 증가하였다. pH는 대조군이 가장 급격한 변화를 보였으며, 식염이 많이 첨가될수록 pH의 변화량이 적었다. 반면, 산도는 pH 감소와 반비례하여 증가하였다. 아미노질소는 식염 5% 첨가군이 가장 짧은 기간에 최고값을 나타내었으며, 식염 10, 15% 첨가군은 식염 5% 첨가군과 비슷한 값을 보였으나 발효기간이 길어 적합하지 않았다. 따라서 이상의 결과를 고려할 때, 식염 5% 첨가군이 조미료를 만들기에 가장 적합하다고 판단하였다. 식염 5%의 농도에 14일간 발효시킨 오징어 내장 발효생성물의 일반성분은 조단백이 가장 많았으며, 유리아미노산 조성은 단맛을 내는 아미노산인 alanine과 감칠맛을 내는 아미노산인 glutamic acid의 함량이 가장 많았다. 항산화와 항대장암 활성은 농도 의존적으로 증가하는 것을 확인하였으나, positive control에 비하여 활성이 떨어졌다. 관능검사 결과는 시판 천연조미료와 비교하여 관능적으로 큰 차이가 없었다. 이상의 결과를 고려할 때 오징어 내장 발효생성물은 천연조미료 등 식품첨가제로써 활용이 가능하다고 판단된다. Squid ( Todarodes pacificus) is processed as dried or seasoned-dried products and its catch gradually increased from 270,298 M/T in 2005 to 367,940 M/T in 2008 in Korea. Squid processing by-product (viscera) was usually discarded as a waste resulting in environmental problem. In order to utilize squid viscera for more value-added products, a natural squid seasoning was developed by fermenting with Aspergillus oryzae koji. Squid viscera at 5, 10 and 15% salt concentrations with fixed levels of 5% koji and 30% water was fermented at room temperature. The quality properties of squid fermented products such as amino-N, TMA, VBN, total viable cell count, pH and total acidity were determined at different fermentation periods. The contents of amino-N, TMA, and VBN of squid seasoning at 5% salt concentration fermented for 14 days were the highest. Based on amino-N content, squid viscera at 5% koji fermented for 14 days was selected for further assays: the content of moisture, crude protein, crude lipid, crude ash, and carbohydrate were 5.98, 35.19, 33.08, 11.30, and 14.45%, respectively. The content of glutamate, alanine, leusine and lysine were 7.06, 12.34, 9.90 and 10.22%, respectively. The IC?? values of DPPH scavenging and β-glucuronidase inhibitory activity were 12.89 and 12.58 ㎎/mL, respectively. A natural squid seasoning was manufactured by mixing fermented squid viscera and an ingredient. Based on the results of sensory evaluation, the fermented squid viscera seasoning was almost equal to other natural complex seasonings such as anchovy, cow meat, and fisheries seasoning.

Accelerating Effect of Squid Viscera on the Fermentation of Alaska pollack Scrap Sauce

Kim, Sang-Moo The Korean Society of Food Science and Nutrition 1999 Preventive Nutrition and Food Science Vol.4 No.2

Fish sauce is a liquid form of salt-fermented fish and has played an important role in Korean dietary life. Fish sauce was manufactured by utilizing Alaska pollack scrap from Himedara(seasoned and dried Alaska pollack tail) processing . In addition, the effects of squid viscera as a fermentation enhancer were also evaluate.Ph of Alaska plllack scrap sauce with squid viscera was lower than that of control over the entire fermentation process. Squid viscera acceleraged the production of amino-nitrogen, VBN , TBA and free amino acids, and the degradation of IMP and Inosine. The addition of squid viscera and koji at 5% concentration, respectively , also accelerated the digestion of Alaska pollack scrap and was similar to the results of squid viscera at 10% concentration.

어유의 효소적 에탄올화 반응 특성

신상규,류홍석,박현덕,전병수,Shin, Sang-Kyu,Yoo, Hong-Suk,Pack, Hyun-Duk,Chun, Byung-Soo 한국해양바이오학회 2006 한국해양바이오학회지 Vol.1 No.4

Enzymatic ethanolysis of fish oil with immobilized lipase was investigated for reducing the free fatty acid contents and enhancing the function of fish oil. Ethanolysis reactions were carried out in erlenmeyer flask (25ml) containing a mixture of squid viscera oil and 99.9% ethanol using 1% (based on w/w squid viscera oil) immobilized lipase. The reaction mixtures were incubated at $50^{\circ}C$ and shaken at 100rpm. Ethanol was added into the mixture by stepwise addition method of Shinmada[9]. Measurement of free fatty acid molar amounts was studied by Acid Value. Tendency of oil variation during transesterification was studied by TLC method. Enzymatic ethanolysis composed diglyceride, monoglyceride and fatty acid ethyl ester with reducing free fatty acid contents. Also, selective ethanolysis by Lipozyme TL-IM and Lipozyme RM-IM mostly did not react at the sn-2 position of squid viscera oil. Lipozyme RM-IM was more suitable enzyme to reduce the free fatty acid contents by ethanolysis than Lipozyme TL-IM. Squid viscera oil was transformed into suitable properties (5 in Acid Value) for functional fish oil production.

초임계 이산화탄소 추출법을 이용한 오징어 가공 부산물로부터 고도불포화 지방산 회수

강성실,김병준,전병수,KANG Seong-Sil,KIM Byung-Jun,CHUN Byung-Soo 한국수산과학회 1999 한국수산과학회지 Vol.32 No.2

A squid viscera oil contains a high content of EPA, DHA, and other valuable polyunsaturated fatty acids. The extractions of squid viscera oil by supercritical carbon dioxide both with/without $3\%$ (v/v) ethanol were performed in a semicontinuous flow extractor at 8.3 to 13.8 MPa and 25 to $50^{\circ}C$. When ethanol was added to $SC-CO_2$, the extraction ratio of lipid increased. The extracts contained high content of unsaturated oils like DHA and EPA. The highest extraction yield of lipid from squid viscera oil extracted by supercritical carbon dioxide was obtained at 12.4 MPa and $40^{\circ}C$ with/without entrainer. The main fatty acids of squid viscera oil extracted by supercritical carbon dioxide were myristric acid (14:0), palmitic acid (16:0), palmitoleic acid (16:1), oleic acid (18:1), arachidic acid (20:0), eicosapentaenoic acid (20:5), and docosahaxaenoic acid (22:6). 초임계 $CO_2$를 이용한 오징어 내장유 추출은 8.3$\~$13.8 MPa과 $25\~50^{\circ}C$에서 semi-continuous flow extractor로 수행되었다. 그리고 초임계 $CO_2$에 entrainer로서의 $2\%$ (v/v) ethanol을 첨가한 것과 첨가하지 않은 것으로 수행되었다. 초임계 $CO_2$를 이용한 오징어 내장유 추출에서 지질의 최고 추출 수율은 조건 12.4 MPa과 $40^{\circ}C$에서 이루어 졌다. 초임계 $CO_2$를 이용한 오징어 내장유 추출에서 최고 용해도는 압력 12.4 MPa $40^{\circ}C$에서 entrainer를 첨가하지 않았을 때 1.48g oil/kg $CO_2$를 나타내었고, entrainer를 첨가하였을 때 압력 12.4MPa $40^{\circ}C$에서 4.85g oil/kg $CO_2$를 나타내었다. 초임계 $CO_2$를 이용하여 추출된 오징어 내장유의 주요지방산조성은 myristric acid (14:0), palmitic acid (16:0), palmitoleic acid (16:1), oleic acid (18:1), arachidic acid (20:0), eicosapentaenoic acid (20:5), docosahexaenoic acid(22:6)이다. 초임계 $CO_2$ 추출에 의해 추출된 추출물중 비소와 납은 검출되지 않으나 수은은 아주 미량으로 검출되었다.