• Title/Summary/Keyword: alcalase

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Purification and Characterization of Antioxidant Peptides from Lotus Nelumbo nucifera Seed Protein (연자육(Lotus Nelumbo nucifera Seed) 단백질로부터 항산화 펩타이드 분리 정제 및 특성)

  • Chathuri K. Marasinghe;Hyun-Woo Kim;Won-Kyo Jung;Jae-Young Je
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.56 no.1
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    • pp.21-27
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    • 2023
  • Lotus Nelumbo nucifera seed protein (LSP) was isolated by alkaline solubilization after removing fat and phenolics by hexane and ethanol treatment. Antioxidant peptides from LSP were produced with Alcalase® and pepsin and hydroxyl radical scavenging activities were determined. LSP-Alcalase® hydrolysates showed higher hydroxyl radical scavenging activity than LSP-pepsin hydrolysates. To purify antioxidant peptides, LSP-Alcalase® hydrolysates were subjected to high performance liquid chromatography (HPLC) separation on the C18 column and the active fraction was further purified using a SuperdexTM peptide 10/300 GL column. Finally, the active fraction (F8-2) was evaluated for antioxidant activities by 2,2-diphenyl-1-picrylhydrazyl (DPPH), hydroxyl radical scavenging, and oxygen radical absorbance capacity (ORAC) assays. The EC50 values of the F8-2 were 105.81±0.02 ㎍/mL for DPPH and 32.26±0.02 ㎍/mL for hydroxyl radical and the F8-2 exhibited 7.22 μM trolox equivalent (TE)/100 ㎍ F8-2. Glutathione (GSH), which is a positive control, showed EC50 values of 19.87±0.01 ㎍/mL for DPPH and 15.95±0.03 ㎍/mL for hydroxyl radical and an ORAC value of 14.17±0.03 μM TE/100 ㎍ GSH. Finally, sixteen peptides were identified by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Among them, Ile-Tyr and Leu-Tyr showed higher antioxidant scores.

Effect of Proteases on the Extraction of Crude Protein and Reducing Sugar in Pollen (화분에서의 조단백질 및 환원당 추출시 단백질 분해효소가 미치는 영향)

  • Choi, Su-Jeong;Jeong, Yoon-Hwa
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.33 no.8
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    • pp.1353-1358
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    • 2004
  • This study was conducted to increase crude protein and reducing sugar contents in pollen extracts by proteases. Four commercial neutral proteases (Alcalase 2.4L, Protamex, Flavozyme and Protease A) and two alkaline proteases (Protease S and Protease P) were used to prepare acorn and Darae pollen extracts. Contents of moisture, ash, crude protein and crude fat of acorn pollen were 5.2%, 2.7%, 6.2% and 22.3%, respectively, while those of Darae pollen were 5.4%, 2.8%, 1.8% and 27.8%, respectively. Contents of crude protein and reducing sugar in pollen extracts were increased by proteases. Alcalase 2.4L was the most effective in increasing protein contents while Protease A in increasing reducing sugar contents. It is suggested the use of proteases is one of the potential methods for increasing the contents of crude protein and reducing sugar in preparation of pollen extracts.

Preparation and Characterization of the Hydrolyzed Protein from Shaving Scraps of Leather Waste Containing Chromium by the Combination Treatment with Alkaline Inducing Agent and Alkaline Proteolytic Enzyme (Alkaline Inducing Agent 및 Alkaline Proteolytic Enzyme 혼용처리에 의한 Shaving Scraps 가수분해 단백질의 제조 및 특성)

  • Kim, Won-Ju;Cho, Ju-Sik;Lee, Hong-Jae;Heo, Jong-Soo
    • Journal of the Korea Organic Resources Recycling Association
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    • v.6 no.1
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    • pp.1-12
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    • 1998
  • To examine the possibility of protein recycling of shaving scraps containing chromium generated from manufacturing process of leather, the optimum hydrolysis conditions and the withdrawal methods of low molecular weight protein for using the liquid fertilizer sources by investigation of solubilities of hydrolyzed protein, inorganic nutrients contents and molecular weight distributions of hydrolyzed protein from shaving scraps treated with mixed alkaline inducing agents and mixed alkaline proteolytic enzymes including MgO were investigated. In hydrolysis of shaving scraps treated with mixed alkaline inducing agents, the solubility of shaving scraps were clearly different with 65~85% according to the sorts of the inducing agents, and the degree of hydrolysis was high in the order of NaOH, $Ca(OH)_2$ and KOH. The average molecular weights of withdrawal hydrolyzed protein were 10, 40 and 80 KD treated with NaOH, $Ca(OH)_2$ and KOH, respectively. And the chromium contents was about 15 ppm. In hydrolysis of shaving scraps treated with mixed alkaline proteolytic enzymes, the bility of shaving scraps were high in the order of alcalase, esperase and savinase. In c of treating 0.5% alcalase, the low molecular weight of hydrolyzed protein could be withdrawn. The solubility of the hydrolyzed protein was about 85%, the average molecular weight of the protein was below 1 KD and chrome content of the protein was below 10 ppm.

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Characteristics of Whey Protein (WPC-30) Hydrolysate from Cheese Whey (치즈유청으로부터 제조한 유청단백질 가수분해물의 특성에 관한 연구)

  • Yoon, Yoh-Chang;An, Sung-Il;Jeong, A-Ram;Han, Song-Ee;Kim, Myeong-Hee;Lee, Chang-Kwon
    • Journal of Animal Science and Technology
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    • v.52 no.5
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    • pp.435-440
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    • 2010
  • Whey protein concentrate (WPC) is widely used to increase the nutritional and functional properties of food. In this study, the physiochemical and functionality of WPC-30 hydrolysates were examined to evaluate the possibility of application in the food industry. The WPC-30 was manufactured using ultrafiltration and spray-drying, and then hydrolyzed with proteolytic enzyme including alcalase, flavourzyme, nuetrase and protamex. Enzymatic hydrolysis had a significant influence on the physicochemical properties as evident from the increased foaming capacity, solubility. Alcalase caused highest protein hydrolysis (3.26%) and the bitterness. Foaming capacity was largest in WPC-30 hydrolysate treated with flavourzyme. Protein solubility at various levels of pH was highest in protamex-treated WPC-30 hydrolysate. However, the solubility of WPC-30 hydrolysates was significantly improved in alkaline condition than in acidic and neutral conditions. The study revealed that spray dried enzyme modified WPC can be used in various functional food.

Extraction and Physicochemical Properties of Collagen from Squid (Todarodes pacificus) skin and Alaska pollack (Theragra chalcogramma) skin (오징어 및 명태껍질 유래 콜라겐의 추출 및 물리화학적 특성)

  • Yang, Su Jin;Hong, Joo-Heon
    • Korean journal of food and cookery science
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    • v.28 no.6
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    • pp.711-719
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    • 2012
  • The collagens from squid (Todarodes pacificus) skin and Alaska pollack (Theragra chalcogramma) skin were extracted and their physicochemical properties were investigated. The yield was improved with the increase of NaOH concentration and was increased Neutrase than Alcalase in enzymatic hydrolysates. Protein and collagen contents from Alaska pollack skin were 38.3~62.7% and 13.1~28.9%, respectively. All enzymatic hydrolysates also showed high antioxidant activities as NaOH concentration decrease. Composition of their amino acids was mainly glycine and proline. The spectrum of FT-IR of the collagen showed wavenumber at $1,631cm^{-1}$, $1,549cm^{-1}$, $1,234cm^{-1}$ and $3,322cm^{-1}$ representing the regions of amide I, amide II, amide III and amide A, respectively. The decomposition temperature for the collagen was in the range of $300^{\circ}C$ and showed relatively good enough for their thermal stabilities.

Angiotensin Ⅰ Converting Enzyme(ACE) Inhibitory Activities of Laver(Porphyra tenera) Protein Hydrolysates (김 단백질 가수분해물의 Angiotensin Ⅰ 전환효소 저해 활성)

  • Kim Young-Myoung;Do Jeong-Ryong;In Jae-Pyung;Park Jong-Hyuk
    • The Korean Journal of Food And Nutrition
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    • v.18 no.1
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    • pp.11-18
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    • 2005
  • Angiotensin Ⅰ converting enzyme(ACE) inhibitory activities of laver(Porphyra tenera) protein hydrolysates were investigated by enzymes used for hydrolysis, molecular fractions and drying methods. For the enzymatic hydrolysis, crude laver protein, separated by filtration of water extract of dried laver extracted with 20 times(w/v) water for 3 hours at boiling temperature, were hydrolyzed with three commercial protease, Pepsin, alcalase and maxazyme NNP at optimal conditions. The yield of hydrolysis and ACE inhibitory activities of which were high in order of pepsin, alcalase and maxazyme NNP. ACE inhibitory activities of laver hydrolysates by molecular levels were high in order of 3 kDa > 10 kDa > 3∼10 kDa, and the IC/sub 50/ ACE inhibitory activities by molecular lebels were 4 mg/mL(3 kDa), 5 mg/mL(total hydrolysate), and 20 mg/mL(10 kDa), respectively. The storage stability of dried laver hydrolysates at 20℃ were strongly affected by drying methods, hot air dried of which were much stabler than freeze-dried one.

Isolation of Iron-Binding Peptides from Sunflower (Helianthus annuus L.) Seed Protein Hydrolysates (해바라기씨박 단백질 가수분해물로부터 철분 결합 펩타이드의 분리)

  • Choi, Dong Won;Kim, Nam Ho;Son, Kyung Bin
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.7
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    • pp.1162-1166
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    • 2013
  • Proteins from sunflower seeds were hydrolyzed with Alcalase and Flavourzyme to isolate iron-binding peptides. The optimal hydrolysis conditions were determined. Hydrolysates were filtered under a 3 kDa membrane and iron-binding peptides separated from the hydrolysates using ion exchange and gel permeation chromatographic methods. A fraction with the highest iron-binding activity (Fe/peptide, 0.69), F22, was obtained. These results suggest that fractions isolated from sunflower seed protein hydrolysates can be applied toward the production of iron supplements.

ACE-inhibitory Effect and Physicochemical Characteristics of Yogurt Beverage Fortified with Whey Protein Hydrolysates

  • Lim, Sung-Min;Lee, Na-Kyoung;Park, Keun-Kyu;Yoon, Yoh-Chang;Paik, Hyun-Dong
    • Food Science of Animal Resources
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    • v.31 no.6
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    • pp.886-892
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    • 2011
  • This study investigated the ACE-inhibitory effect of yogurt beverage fortified with hydrolysates as well as the suitability of hydrolysates as a nutraceutical additive to yogurt beverage. Three whey protein hydrolysates hydrolyzed by alcalase, protamex, and trypsin were each added to yogurt beverage at concentrations of 1.25, 2.5, and 5 mg/mL. Yogurt beverage fortified with 2.5 mg/mL of hydrolysates had 61-69% ACE-inhibitory activity, whereas yogurt beverage fortified with 5 mg/mL of hydrolysates showed 74% ACE-inhibitory activity. There were no significant differences in ACE-inhibitory activity between the alcalase or protamex hydrolysates during storage; however, trypsin hydrolysate exhibited significant differences. On the other hand, physicochemical characteristics such as pH (3.47-3.77), titratable acidity (0.81-0.84%), colority, viable cell count, and sensory qualities were not significantly different among the tested yogurt beverage samples during storage. These results showed that yogurt beverage fortified with whey protein hydrolysates maintained antihypertensive activity and underwent no unfavorable changes in physicochemical characteristics regardless of enzyme type.

Hydrolysis Conditions of Porcine Blood Proteins and Antimicrobial Effects of Their Hydrolysates

  • Jin, Sang Keun;Choi, Jung Seok;Yim, Dong-Gyun
    • Food Science of Animal Resources
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    • v.40 no.2
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    • pp.172-182
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    • 2020
  • In the present study, we determined the degree of hydrolysis (DH) of porcine blood plasma proteins, albumin, and globulin hydrolyzed by six proteases (alcalase, neutrase, flavourzyme, protamex, trypsin, and papain) for various reaction times. Moreover, antimicrobial activities of hydrolysates against five pathogenic microorganisms (Bacillus cereus, Staphylococcus aureus, Salmonella Typhimurium, Escherichia coli, and Shigella flexneri) were investigated. Alcalase, trypsin, and papain hydrolysates of the three porcine blood proteins showed higher DH values than hydrolysates produced by the other three proteases. DH of the three porcine blood proteins hydrolyzed by the six proteases failed to increase after 2 h of hydrolysis. In antimicrobial tests, hydrolysates (hydrolysis time of 2 h) showed antibacterial activity only against B. cereus. Albumin hydrolysates showed higher antimicrobial activity than globulin and plasma hydrolysates. Albumin hydrolysates obtained with flavourzyme, protamex, and trypsin showed higher antimicrobial activity than those obtained with the other three proteases.

Potential Antioxidant Activities of Enzymatic Digests from Fresh Water Microalgae, Pediastrum duplex and Dactylococcopsis fascicularis

  • Lee, Seung-Hong;Kim, Areum-Daseul;Kang, Min-Cheol;Lee, Joon-Baek;Jeon, You-Jin
    • ALGAE
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    • v.24 no.3
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    • pp.169-177
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    • 2009
  • In this study, we focused on natural water-soluble antioxidants from fresh water microalgae, Pediastrum duplex and Dactylococcopsis fascicularis from Jeju Island, Korea. They were prepared by enzymatic digestion using five carbohydrases (Viscozyme, Celluclast, AMG, Termamyl and Ultraflo) and five proteases (Protamex, Kojizyme, Neutrase, Flavourzyme and Alcalase), and the potential antioxidant activity of each was assessed. All enzymatic digests from P. duplex showed significant DPPH scavenging effects. Termamyl (60.6%) digest from P. duplex possessed the highest effects on hydrogen peroxide scavenging. Celluclast (58.1%) and Kojizyme digests (56.9%) from D. fascicularis possessed higher effects on superoxide anion radical scavenging. All enzymatic digests exhibited significant effects on both NO· scavenging and metal chelating. Lipid peroxidation was significantly in inhibited Viscozyme, Termamyl and Kojizyme digests from P. duplex and Ultraflo, Protamex, Kojizyme and Alcalase digests from D. fascicularis. These data suggest that enzymatic digests of the fresh water microalgae, P. duplex and D. fascicularis might be valuable sources of antioxidant which can be applied in food and pharmaceutical industry.