• 제목/요약/키워드: enzyme hydrolysis

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Optimization of fish oil extraction from Lophius litulon liver and fatty acid composition analysis

  • Hu, Zhiheng;Chin, Yaoxian;Liu, Jialin;Zhou, Jiaying;Li, Gaoshang;Hu, Lingping;Hu, Yaqin
    • Fisheries and Aquatic Sciences
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    • v.25 no.2
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    • pp.76-89
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    • 2022
  • The Lophius litulon liver was used as raw material for the extraction of fish oil via various extraction methods. The extraction rate by water extraction, potassium hydroxide (KOH) hydrolysis and protease hydrolysis were compared and the results revealed the protease hydrolysis extraction had a higher extraction rate with good protein-lipid separation as observed by optical microscope. Furthermore, subsequent experiments determined neutrase to be the best hydrolytic enzyme in terms of extraction rate and cost. The extraction conditions of neutrase hydrolysis were optimized by single-factor experiment and response surface analysis, and the optimal extraction rate was 58.40 ± 0.25% with the following conditions: enzyme concentration 2,000 IU/g, extraction time 1.0 h, liquid-solid ratio 1.95:1, extraction temperature 40.5℃ and pH 6.5. The fatty acids composition in fish oil from optimized extraction condition was composed of 19.75% saturated fatty acids and 80.25% unsaturated fatty acids. The content of docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) were 8.06% and 1.19%, respectively, with the ratio (6.77:1) surpassed to the recommendation in current researches (5:1). The results in this study suggest protease treatment is an efficient method for high-quality fish oil extraction from Lophius litulon liver with a satisfactory ratio of DHA and EPA.

Quality Characteristics of 4 Year-old Ginseng by Enzymatic Hydrolysis Conditions (4년근 인삼의 효소적 가수분해 조건에 따른 품질특성)

  • Im, Ga-Young;Ma, Jin-Yeul;Kim, Kun-Woo;Choi, Jin-Kook;Kang, Dong-Kyoon;Kwon, Tae-Ryoung;Jang, Se-Young;Jeong, Yong-Jin
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.2
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    • pp.229-234
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    • 2011
  • This study investigated quality characteristics of 4 year-old ginseng by enzymatic hydrolysis conditions to increase utilization. Ginseng was ground after steaming and was each treated with hydrolase A, B, C and D. When quality characteristics by the enzymes were examined, no significant difference was observed with pH of 5.5~5.6 and the sugar content of 4.0~4.33. The crude saponin content was the highest in ginseng treated with D, followed by B, C and A. The crude saponin, the reducing sugar and the total sugar contents increased until 0.3% (w/w) concentration in enzyme D with no significant difference by its concentration. Although active ingredients increased with time passage of hydrolysis, no significant change was found after three hours and the crude saponin content was the highest when ginseng was treated at $60^{\circ}C$. From these results, optimum conditions for 4 year-old ginseng were $60^{\circ}C$ for 3 hours with 0.3% (w/w) enzyme D, and under these conditions the reducing sugar, the total sugar and the crude saponin contents recorded 18.11, 36.21 and 4.23 mg/g, respectively. Therefore, enzymatic hydrolysis was found to be effective in increasing active ingredients of 4 year-old ginseng with various usages expected.

Cloning and Expression in Escherichia coli of a Bacteriolytic Enzyme Gene from Alkalophilic Bacillus sp.

  • Yu, Ju-Hyun;Jung, Myeong-Ho;Park, Hee-Kyoung
    • Journal of Microbiology and Biotechnology
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    • v.2 no.3
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    • pp.161-165
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    • 1992
  • The gene encoding the bacteriolytic enzyme cell wall peptidoglycan hydrolase from alkalophilic Bacillus sp. was cloned in E. coli using pBR322 as a vector. A recombinant plasmid, designated pYTR451, was isolated and the size of the cloned HindIII fragment was found to be 4.8 Kb. The cell wall hydrolysis activity of an extract of the E. coli harboring the recombinant plasmid pYTR 451 was detected by SDS- polyacrylamide gel containing 0.2% (w/v) purified cell wall of Bacillus sp. The molecular weight of the enzyme was estimated to be about 27, 000 corresponding to the molecular weight of the Bacillus sp. bacteriolytic enzyme. The recombinant plasmid was found to contain the fragment originated from Bacillus sp. YJ-451 chromosomal DNA by Southern hybridization.

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Effect of ${\alpha}-Amylase$ Treatment of Brown Rice(Goami) Alcohol Fermentation By-Product (현미(고아미) 알코올발효 부산물의 ${\alpha}-amylase$처리 효과)

  • Woo, Seung-Mi;Kim, Tae-Young;Yeo, Soo-Hwan;Kim, Sang-Burm;Kim, Mi-Hyun;Woo, Sang-Chel;Jeong, Yong-Jin
    • Food Science and Preservation
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    • v.14 no.6
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    • pp.617-623
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    • 2007
  • To utilize the non-heat treated alcoholic by-products of brown rice(Goami) as food sources, the quality characteristics change according to the treatment conditions of ${\alpha}-amylase$ were evaluated. It resulted that the increase of hydrolysis temperature correspondingly increased the soluble solids, total dietary fiber and total sugar in the by-products of Goami, and the highest reducing sugar content was observed at $80^{\circ}C$. The free amino acids contents were tended to slowly decrease by the hydrolysis temperature more than $70^{\circ}C$, and the highest content of oligosaccharides were detected at the hydrolysis temperature of $80^{\circ}C$. The soluble solid according to the ${\alpha}-amylase$ concentration resulted to increase with the increase of the enzyme concentration and the total dietary fiber revealed similarly showing approximately 0.65%. The high content of reducing sugars was observed at the enzyme concentration around 0.08%(v/w). Total sugars and oligosaccharides contents tend to increase as the concentration of enzyme increased, and the content of oligosaccharides acquired at the enzyme concentration more than 0.10%(v/w) maintained to show rather similar contents. The soluble solids and total dietary fiber by hydrolysis time were found to show 6.66% and 0.65%, respectively at more than 60 min of hydrolysis, and the reducing sugars and total sugars were found to be 3,600 and 4,800 mg% in all treatment groups showing no significant difference. The content of oligosaccharides was increased with the increase of hydrolysis time, and the content was similar at more than 90 min of hydrolysis by ranging around 2,100 mg%. Based upon these results, the by-products of Goami are expected to be used as various food sources showing the highest dietary fiber and oligosaccharides contents by the hydrolysis at $80^{\circ}C$ for 90 min with the addition of 0.10%(v/w) of ${\alpha}-amylase$.

Development of Optimum Process for Continuous Hydrolysis of Fish Skin Gelatin Using a Three-Step Recycle Membrane Reactor (재순환 3단계 막반응기를 이용한 어피젤라틴의 연속적 가수분해 최적화 공정 개발)

  • Kim, Se-Kwon;Byun, Hee-Guk
    • Applied Chemistry for Engineering
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    • v.5 no.4
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    • pp.681-697
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    • 1994
  • The enzymatic hydrolysate of gelatin extracted from fish skin was fractionated and recycled through the membrane reactor according to the molecular weight for the purpose of using as functional material. In addition, the enzymatic hydrolysis conditions of gelatin, enzyme stability by membrane and mechanical shear, and effect on the long-term operational stability of the recycle membrane reactor were investigated. Using the pH-drop technique, Alcalase, pronase E and collagenase were identified as the most suitable enzymes for the hydrolysis of fish skin gelatin. The optimum hydrolysis conditions in the 1st-step membrane reactor(1st-SMR) by Alcalase were enzyme concentration 0.2mg/ml, substrate-to-enzyme ratio(S/E) 50(w/w), $50^{\circ}C$, pH 8.0, reaction volume 600ml and flow rate 6.14ml/min. In the 2nd-SMR by pronase E were enzyme concentration 0.3mg/ml, S/E 33(w/w), $50^{\circ}C$, pH 8.0, reaction volume 600ml and flow rate 6.14ml/min. In the case of 3rd-SMR, enzyme concentration 0.1mg/ml, S/E 100(w/w), $37^{\circ}C$, pH 7.5, reaction volume 600ml and flow rate 10ml/min. Decreased enzyme activities by mechanical shear and membrane were 30% and 15% in the 1st-SMR, were 14% and 5% in the 2nd-SMR, and 18% and 8% in the 3rd-SMR, respectively. Under the optimum conditions, the degree of hydrolysis in the 1st, 2nd and 3rd-SMR were 3.5%(Kjeldahl method, 87%), 3.1%(77%) and 2.7%(70%), respectively. The productivity of hydrolysate in the continuous three-step membrane reactor was 430mg per enzyme(mg) for 10 times of volume replacements.

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Hydrolysis of Castor Oil with Lipases and Organic Solvents (Lipase와 유기용매를 이용한 Castor Oil의 가수분해)

  • Jeon, Gyu-Jong;Hur, Byung-Ki;Yang, Ji-Won
    • KSBB Journal
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    • v.14 no.6
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    • pp.696-701
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    • 1999
  • The enzymatic hydrolysis of Castor oil for the mass production of ricinoleic acid was studied to find out the optimum conditions such as solvents and the weight ratio of substrate to enzyme. Three different lipases were tested for the hydrolysis of castor oil: lipase from Porcine Pancrease(lipsase PP), lipase from Candida cylindracea(lipase CC), lipase from Candida Rugosa(lipase CR). The poor mass transfer in water caused a low degree of hydrolysis of castor oil. To overcome this problem, organic solvents were used. Among organic solvents tested, hydrophobic solvents gave better results of hydrolysis than hydrophilic solvents. Organic solvents also lowered or changed the effect of pH. Isopropyl ether made complete hydrolysis of castor oil. The ratio of water to isopropyl ether and the ratio of weight ratio of lipase to castor oil were important for the hydrolysis of castor oil. At 30$^{\circ}C$ castor oil was completely hydrolyzed by 4 wt% of lipase in the mixture of isopropyl ether and water(1:1 in volume).

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The Hydrolysis Conditions of Rapeseed Protein by Pronase (유채단백질의 단백효소에 의한 가수분해 조건)

  • Kim, Chung-Hee;Kim, Hyo-Sun;Jung, Yong-Hyun;Kang, Yeung-Joo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.21 no.5
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    • pp.513-518
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    • 1992
  • Optimum conditions for enzymatic hydrolysis of purified rapeseed(Brassica napus var. Youngsan) protein were investigated. Pronase showed higher activity in the hydrolysis of rapeseed protein than that of alcalase and neutrase. Preheated treatment of the rapeseed protein decreased the activity of pronase. The degree of hydrolysis of rapeseed protein was greater in distilled water than in phosphate buffer solution. Degree of hydrolysis was reached in steady state after 1 hr. Optimum conditions of the hydrolysis of the rapeseed protein were $40^{\circ}C$ in reaction temperaturem pH 8.0 in substrate solution, 1/100 (w/w) in the ratio of enzyme to substrate and 1% (w/v) in substrate concentration for pronase, respectively. At the optimum hydrolysis conditions, Km value was 3.48% (w/v).

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Bioethanol Production from Wasted Corn Stalk from Gangwon Province : from Enzymatic Hydrolysis to Fermentation (강원지역 폐옥수수대로부터 바이오에탄올 생산 : 효소 당화부터 발효까지)

  • Choi, Jae Min;Choi, Suk Soon;Yeom, Sung Ho
    • Applied Chemistry for Engineering
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    • v.23 no.3
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    • pp.326-332
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    • 2012
  • Among the samples prepared by various pre-treatment methods, the one pretreated by dilute sulfuric acid showed the highest glucose yield in the enzymatic hydrolysis. Statistical analysis of enzymatic hydrolysis revealed that the glucose yield was in proportion to the enzyme dosage, the ratio of the pre-treated sample to the buffer solution, and the reaction time and that the effect of enzyme dosage was predominant in the experiment range. In addition, the glucose yield was estimated to be 76.1% at an optimal enzymatic hydrolysis condition. In a separate hydrolysis and fermentation (SHF), Saccharomyces cerevisiae converted over 80% of glucose from the enzymatic hydrolysis of pre-treated wasted corn stalk by dilute sulfuric acid to bioethanol with 37% of ethanol yield and 0.42 $g/L{\cdot}hr$ of productivity. In the simultaneous saccharification and fermentation (SSF), 59.5% of conversion from glucan to ethanol and 0.20 $g/L{\cdot}hr$ of productivity were achieved. In both SHF and SSF, approximately 88 g of bioethanol could be obtained from 1 kg of wasted corn stalk. The possible amount of bioethanol in Gangwon province were estimated to be 1.9 kiloton with the assumption of the 50% of collection ratio.

Purification and Characterization of D-Xylanase II from Penicillium verruculosum (Penicillium verruculosum으로부터 D-xylanase II의 정제 및 특성)

  • 조남철;강영태;이태훈;정기철;김강화
    • Microbiology and Biotechnology Letters
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    • v.21 no.6
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    • pp.588-593
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    • 1993
  • Xylanase(1, 4-beta-D-xylan xylanohydrolase` EC 3.2.1.8) II was purified from Penicillium verruculosum by using the techniques of two anion exchange chromatographies, and gel filtration. The molecular weight of this enzyme was about 22, 000 as determined by SDS-electrophoresis. The enzyme showed hydropytic activity toward xylan but did not catalyze hydrolysis of Rho-nitrophenyl-beta-D-xylopyranoside, Rho-nitrophenyl-beta-D-glucopyranoside, Rho-nitrophenyl-beta-D-cellobiopyranoside, and celluloses such as Avicel, cotton, filter paper, carboxymethylcellulose.

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Saprolegnia ferax에 의한$\beta$-amylase의 생산 및 특성

  • Bai, Suk;Cho, Nam-Chul;Chun, Soon-Bai
    • Microbiology and Biotechnology Letters
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    • v.25 no.2
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    • pp.109-114
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    • 1997
  • The Oomycete Saprolegnia ferax produces an extracellular $\beta$-amylase, Maximum enzyme yield was attained after 7 days of growth in YNB starch medium (pH 6.5) at 25$\circ$C. The amylase was pu- rified 24-fold by ultrafitration, HPLC DEAE column and HPLC gel filtration. The purfied enzyme was a monomeric glycoprotein with a molecular weight of about 44,000 dalton. The pH and temperature optima were 6.5 and 50$\circ$C, respectively. The enzyme was fairly stable up to 50$\circ$C and at acidic pH region (pH 4.0-7.0). The apparent Km and Vmax values of the enzyme against soluble starch were 0.77 mg/ml and 2,174 $\mu$moles/mg protein, respectively. Amino acid analysis indicated that the enzyme was enriched in alanine, glycine, leucine and acidic amino acid. Starch hydrolysis with the enzyme released maltose but not glucose, whereas maltotriose, Schardinger dextrin ($\alpha$-cyclodextrin) and pullulan were not hydrolysed by the enzyme. The enzyme was inhibited by Schardinger dextrin, p-chloromercuribenzoate(PCMB), CU$^{2+}$' and Hg$^{2+}$. Inhibition of the enzyme by PCMB could be reversed by the addition of cysteine and mercaptoethanol.

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