• Title/Summary/Keyword: Proteolytic processing

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Effects of fermentation on protein profile of coffee by-products and its relationship with internal protein structure measured by vibrational spectroscopy

  • Samadi;Xin Feng;Luciana Prates;Siti Wajizah;Zulfahrizal;Agus Arip Munawar;Peiqiang Yu
    • Animal Bioscience
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    • v.36 no.8
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    • pp.1190-1198
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    • 2023
  • Objective: To our knowledge, there are few studies on the correlation between internal structure of fermented products and nutrient delivery from by-products from coffee processing in the ruminant system. The objective of this project was to use advanced mid-infrared vibrational spectroscopic technique (ATR-FT/IR) to reveal interactive correlation between protein internal structure and ruminant-relevant protein and energy metabolic profiles of by-products from coffee processing affected by added-microorganism fermentation duration. Methods: The by-products from coffee processing were fermented using commercial fermentation product, called Saus Burger Pakan, consisting of various microorganisms: cellulolytic, lactic acid, amylolytic, proteolytic, and xylanolytic microbes, for 0, 7, 14, 21, and 28 days. Protein chemical profiles, Cornell Net Carbohydrate and Protein System crude protein and CHO subfractions, and ruminal degradation and intestinal digestion of protein were evaluated. The attenuated total reflectance-Ft/IR (ATR-FTIR) spectroscopy was used to study protein structural features of spectra that were affected by added microorganism fermentation duration. The molecular spectral analyses were carried using OMNIC software. Molecular spectral analysis parameters in fermented and non-fermented by-products from coffee processing included: Amide I area (AIA), Amide II (AIIA) area, Amide I heigh (AIH), Amide II height (AIIH), α-helix height (αH), β-sheet height (βH), AIA to AIIA ratio, AIH to AIIH ratio, and αH to βH ratio. The relationship between protein structure spectral profiles of by-products from coffee processing and protein related metabolic features in ruminant were also investigated. Results: Fermentation decreased rumen degradable protein and increased rumen undegradable protein of by-products from coffee processing (p<0.05), indicating more protein entering from rumen to the small intestine for animal use. The fermentation duration significantly impacted (p<0.05) protein structure spectral features. Fermentation tended to increase (p<0.10) AIA and AIH as well as β-sheet height which all are significantly related to the protein level. Conclusion: Protein structure spectral profiles of by-product form coffee processing could be utilized as potential evaluators to estimate protein related chemical profile and protein metabolic characteristics in ruminant system.

Overview of Milk Allergens and Allergic Reaction Reduction Methods (우유 알레르기의 특성 및 저감화 방법에 대한 고찰)

  • Kim, Ki-Hwan;Seol, Kuk-Hwan;Oh, Mi-Hwa;Park, Beam Young;Kim, Hyoun Wook
    • Journal of Dairy Science and Biotechnology
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    • v.31 no.1
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    • pp.67-73
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    • 2013
  • Food allergy is defined as adverse reactions toward food mediated by aberrant immune mechanisms. Cow's milk allergy is one of the most common food allergies in childhood. This allergy is normally outgrown in the first year of life, however 15% of allergic children remain allergic. Cow's milk allergy seem to be associated with casein (${\alpha}_{s1}$-CN), ${\beta}$-lactoglobulin and whey protein. In addition to this, many other milk proteins are antigenic and capable of inducing immune responses. Various food processing affects the stability, structure and intermolecular interactions of cow milk proteins, as a result reduction the allergenic capacity. Heating, hydrolysis, chemical, proteolytic and other processes such as gamma-ray irradiation, high pressure, using probiotics treatments of milk to obtain hypoallergenic milk have been developed to reduce allergic reactions.

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An Automated High Throughput Proteolysis and Desalting Platform for Quantitative Proteomic Analysis

  • Arul, Albert-Baskar;Han, Na-Young;Lee, Hookeun
    • Mass Spectrometry Letters
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    • v.4 no.2
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    • pp.25-29
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    • 2013
  • Proteomics for biomarker validation needs high throughput instrumentation to analyze huge set of clinical samples for quantitative and reproducible analysis at a minimum time without manual experimental errors. Sample preparation, a vital step in proteomics plays a major role in identification and quantification of proteins from biological samples. Tryptic digestion a major check point in sample preparation for mass spectrometry based proteomics needs to be more accurate with rapid processing time. The present study focuses on establishing a high throughput automated online system for proteolytic digestion and desalting of proteins from biological samples quantitatively and qualitatively in a reproducible manner. The present study compares online protein digestion and desalting of BSA with conventional off-line (in-solution) method and validated for real time sample for reproducibility. Proteins were identified using SEQUEST data base search engine and the data were quantified using IDEALQ software. The present study shows that the online system capable of handling high throughput samples in 96 well formats carries out protein digestion and peptide desalting efficiently in a reproducible and quantitative manner. Label free quantification showed clear increase of peptide quantities with increase in concentration with much linearity compared to off line method. Hence we would like to suggest that inclusion of this online system in proteomic pipeline will be effective in quantification of proteins in comparative proteomics were the quantification is really very crucial.

Recovery of Bioavailable Calcium from Alaska Pollack (Theragra chalcogramma) Fish Backbone By-products by Pepsinolytic Hydrolysis

  • Karawita Rohan;Heo, Soo-Jin;Lee, Bae-Jin;Kim, Se-Kwon;Song, Choon-Bok;Jeon, You-Jin
    • Preventive Nutrition and Food Science
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    • v.11 no.2
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    • pp.120-126
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    • 2006
  • Fish backbone, a major by-product in the fish processing industry, accounts for about 15% of whole fish weight. In this study, recovery of bioavailable calcium from Alaska pollack (Theragra chalcogramma) backbone by-products using enzymatic hydrolysis was investigated. Finely ground fish backbones were hydrolyzed with two proteolytic enzymes (pepsin and protease) to obtain soluble calcium from the by-products. The pepsin digest had a higher degradation efficiency (88%) than protease. Four different concentrations of the fish backbone calcium (100, 250, 500 and 1000 mg/L) prepared by the pepsin digest were treated with $Na_2HPO_4$ at a concentration gradient (0, 1, 2, 4, 8, 10, 15 and 20 mM) to evaluate their solubility, revealing that solubilities of the fish backbone calcium were superior to those of $CaCl_2$ at all the calcium and $Na_2HPO_4$ concentrations. Among the tested concentrations the highest solubility was found in the pepsin digest containing a calcium concentration of 1000 mg/L. Thus, hydrolyzing with pepsin is an effective mode of recovering bioavailable calcium from Alaska pollack fish backbones.

Processing Conditions of Low-Salt Fermented Squid and Its Flavor Components 3. Characterization of Protease Produced by Pseudomonas D2 Isolated from Squid Jeotkal (저염 오징어젓갈 제조 방법 및 향미 성분 3. 오징어젓갈에서 분리한 Pseudomonas D2가 생성하는 Protease의 효소학적 특성)

  • 허성호;이호재;김형선;최성희;김영만
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.24 no.4
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    • pp.636-641
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    • 1995
  • Proteolytic activities were compared using three species involving in squid jeotkal fermentation and showing positive reaction upon casein test : Pseudomonas D2, Flavovacterium odoratum and Acinetobacter calcoaceticus. Pseudomonas D2 produced highest activity of protease at 72h when incubated in our own modified medium(polypeptone, 0.5% ; tryptone, 0.5% ; NaCl, 3% ; pH, 7.5). Thus, this specie was selected for the further study. The growth pattern was coincided with the production of protease. Thus purification of protease was proceeded by ethanol precipitation, sephadex G-100 gel filtration, and DEAE sepharose ion exchange chromatography. The purified protease showed highest activity at pH 7.0 and 5$0^{\circ}C$. The enzyme was very stable over the wide ragnes of the temperature ; even with one hour heat treatment at 7$0^{\circ}C$, the enzyme showed substantial amount of the activity toward casein. In addition, the enzyme was stable over the wide range of pH. Molecular weight of the protease was determined to be 17.4 kD by SDS-PAGE.

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Effects of Heat Treatment on the Nutritional Quality of Milk: V. The Effect of Heat Treatment on Milk Enzymes (우유의 열처리가 우유품질과 영양가에 미치는 영향: V. 열처리가 우유효소에 미치는 영향)

  • Shin, Hanseob;Oh, Sejong
    • Journal of Dairy Science and Biotechnology
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    • v.36 no.1
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    • pp.49-71
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    • 2018
  • Heat treatment is the most popular processing technique in the dairy industry. Its main purpose is to destroy the pathogenic and spoilage bacteria in order to ensure that the milk is safe throughout its shelf life. The protease and lipase that are present in raw milk might reduce the quality of milk. Plasmin and protease, which are produced by psychrotrophic bacteria, are recognized as the main causes of the deterioration in milk flavor and taste during storage. The enzymes in raw milk can be inactivated by heat treatment. However, the temperature of inactivation varies according to the type of enzyme. For example, some Pseudomonas spp. produce heat-resistant proteolytic and lipolytic enzymes that may not be fully inactivated by the low temperature and long time (LTLT) treatment. These types of enzymes are inhibited only by the high temperature and short time (HTST) or ultra-high temperature (UHT) treatment of milk.

Studies on the Processing of Accelerated Low Salt-Fermented Anchovy Paste by Adding Koji (Koji를 이용한 저식염 멸치젓의 속성제조에 관한 연구)

  • 차용준;김은정;주동식
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.23 no.2
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    • pp.348-352
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    • 1994
  • The adding effect of Koji, made by Aspergillus oryzae, in low salt-fermented anchovy paste was studied to reduced fermentation time and to enhance flavor quality as a commercial feasibility in seafood industry. The content of volatile basic nitrogen increased slowly during fermentation , whereas that of amino nitrogen was the highest in 30 days of low salt-fermented anchovy made with Koji. Protease and lipase activity in anchovy paste, which made with or without Koji, were high amount in 30 and 20 days of fermentation, respectively. A number of proteolytic bacteria was high level in 13 days and then slowly decreased afterward.

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In Vitro Formation of Active Carboxypeptidase Y from Pro-Carboxypeptidase Y Inclusion Bodies by Fed-Batch Operation

  • Hahm, Moon-Sun;Chung, Bong-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.11 no.5
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    • pp.887-889
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    • 2001
  • The gene encoding yeast pro-carboxypeptidase Y (pro-CPY) has been cloned and expressed in Escherichia coli. Most of the expressed pro-CPY was accumulated as cytoplasmic insoluble aggregates. In our previous study, active CPY was obtained by renaturation of entirely denatured pro-CPY followed by in vitro proteolytic processing with proteinase K along with the activation process. The same refolding process was performed to produce an active CPY from pro-CPY inclusion bodies with renaturation buffers containing proteinase K at different concentrations. The refolding efficiency decreased from $25\%\;to\;2\%$ in the renaturation buffers containing proteinase K at concentrations of $60{\mu}g/ml\;and\;0.6{\mu}g/mi$, respectively. In an attempt to increase the refolding efficiency with a lesser amount of proteinase K, a novel fed-batch refolding process was developed. In a fed-batch refolding, 99 ml of the renaturation buffer containing pro-CPY was gradually added into 1 ml of the renaturation buffer containing $60{\mu}g/ml$ of proteinase K to give a final proteinase K concentration of $0.6{\mu}g/ml$. The fed-batch refolding process resulted in a refolding efficiency of $18\%$, which corresponded to a 9-fold increase over that ($2\%$) in the batch process.

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Optimal Condition of Natural Silk 3D Matrix Production by Silkworm Spinning

  • Bae, Sung Min;Kweon, HaeYong;Jo, You-Young
    • International Journal of Industrial Entomology and Biomaterials
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    • v.35 no.2
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    • pp.83-88
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    • 2017
  • Silk is appealing materials for many biomedical applications involving tissue engineering and implantable devices, because of its biocompatibility, environmental stability, controlled proteolytic biodegradability and morphologic flexibility. Silk matrix is required for the treatment of a wide wound area, but the present silk matrix is made by the second processing, and thus, the labor and the cost are high. In this work, we investigated the optimal production condition of natural silk 3D matrix using the silkworms and invented Automatic Silk Matrix Making Machine (ASMMM) for natural silk 3D matrix production. As a result, we determined that optimal production condition for making A4 paper size natural silk 3D matrix was used Rough aquarelle paper on surface at $25^{\circ}C$ and 30 silkworm larvae. These results are expected to provide basic data for the efficient production of the natural silk 3D matrix, and it is suggested that the produced natural silk 3D matrix is useful as a medical biomaterials.

Potential role of ANGPTL4 in cancer progression, metastasis, and metabolism: a brief review

  • Min Seok Park;Sang Eun Kim;Pureunchowon Lee;Ju-Hee Lee;Kyung Hee Jung;Soon-Sun Hong
    • BMB Reports
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    • v.57 no.8
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    • pp.343-351
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    • 2024
  • Angiopoietin-like 4 (ANGPTL4) has been identified as an adipokine involved in several non-metabolic and metabolic diseases, including angiogenesis, glucose homeostasis, and lipid metabolism. To date, the role of ANGPTL4 in cancer growth and progression, and metastasis, has been variable. Accumulating evidence suggests that proteolytic processing and posttranslational modifications of ANGPTL4 can significantly alter its function, and may contribute to the multiple and conflicting roles of ANGPTL4 in a tissue-dependent manner. With the growing interest in ANGPTL4 in cancer diagnosis and therapy, we aim to provide an up-to-date review of the implications of ANGPTL4 as a biomarker/oncogene in cancer metabolism, metastasis, and the tumor microenvironment (TME). In cancer cells, ANGPTL4 plays an important role in regulating metabolism by altering intracellular glucose, lipid, and amino acid metabolism. We also highlight the knowledge gaps and future prospect of ANGPTL4 in lymphatic metastasis and perineural invasion through various signaling pathways, underscoring its importance in cancer progression and prognosis. Through this review, a better understanding of the role of ANGPTL4 in cancer progression within the TME will provide new insights into other aspects of tumorigenesis and the potential therapeutic value of ANGPTL4.