• Title/Summary/Keyword: alternative protein

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Enhancement of Glucose-Fueled Cell-Free Protein Synthesis by the Addition of Lipids (지질의 첨가를 통한 포도당 기반 무세포 단백질 합성 시스템의 단백질 발현 효율 향상)

  • Lee, So Jeong;Kim, Ho-Cheol;Kim, Dong-Myung
    • Korean Chemical Engineering Research
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    • v.57 no.1
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    • pp.85-89
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    • 2019
  • Cell-free protein synthesis utilizes the translational machinery in a cell extract. Unlike the conventional cell-based expression methods, not being affected by the conditions for cell growth, cell-free protein synthesis enables flexible manipulation of individual factors affecting the efficiency protein biosynthesis. However, the high cost and low stability of the energy sources to regenerate ATP have limited the use of cell-free synthesis for large-scale production of recombinant proteins. One of the approaches to address this problem is to use glucose as an alternative energy source to regenerate ATP through the glucose-metabolizing pathways in a cell extract. In this study, in an attempt to improve the efficiency of ATP regeneration by reinforcing oxidative phosphorylation process, we supplemented with cellular lipids to a glucose-fueled reaction mixture for cell-free protein synthesis. As a result of the lipid supplementation, the productivity of chloramphenicol acetyltransferase in a cell-free synthesis system using glucose increased more than 6 fold compared to when the lipid was not supplemented.

Inhibition of lipid and protein oxidation in raw ground pork by Terminalia arjuna fruit extract during refrigerated storage

  • Chauhan, Pranav;Pradhan, Soubhagya Ranjan;Das, Annada;Nanda, Pramod Kumar;Bandyopadhyay, Samiran;Das, Arun K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.2
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    • pp.265-273
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    • 2019
  • Objective: Terminalia arjuna plant, specially its leaves, bark, and roots, are widely used in traditional herbal medicine due to presence of bioactive components and being a rich source of natural antioxidants. But its fruit has not been used for any such purposes despite its potential to retard oxidation. Hence, the antioxidant potential of Arjuna fruit extract (AFE) in retarding lipid and protein oxidation of raw ground pork was evaluated during refrigerated storage for 9 days. Methods: The AFEs were prepared using different solvents viz. ethanol (EH), water, ethanol: water (60:40) and methanol:hot water (60:40). The AFEs were analysed for total phenolic content (TPC), 2, 2-diphenyl-1-picrylhydrazyl radical scavenging activity and reducing power. Water extract (WE) and ethanol-water extract (EH-WE) were selected and incorporated at 1.0% into freshly minced pork meat and compared with a synthetic antioxidant, in retarding lipid and protein oxidation during storage. Results: The TPC in AFEs using different solvents ranged from 11.04 to 16.53 mg gallic acid equivalents/g and extracts exhibited appreciable scavenging activity ranging from 50.02% to 58.62%. Arjuna extracts significantly (p<0.05) improved the colour score of meat samples by reducing the formation of metmyoglobin during storage. Both the AFEs (WE and EH-WE) significantly (p<0.05) lowered the thiobarbituric acid reactive substances value, peroxide formation and formation of protein carbonyls in raw pork than control sample during storage. Upon sensory evaluation of all samples, it was found that AFE treatment could prolong the storage period of meat samples, without influencing the colour and odour score, up to 6 days. Conclusion: AFEs used at 1% improved the oxidative stability, colour and odour score and prolonged the refrigerated shelf life of ground pork up 6 days. Therefore, AFE could be explored as an alternative natural antioxidant in retarding lipid and protein oxidation in meat products.

Effects of Blanching Methods on Nutritional Properties and Physicochemical Characteristics of Hot-Air Dried Edible Insect Larvae

  • Jae Hoon Lee;Tae-Kyung Kim;Sun-Young Park;Min-Cheol Kang;Ji Yoon Cha;Min-Cheol Lim;Yun-Sang Choi
    • Food Science of Animal Resources
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    • v.43 no.3
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    • pp.428-440
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    • 2023
  • Global meat consumption is increasing worldwide, however, supply remains lacking. Several alternative protein sources, such as cultured meat, plant-based protein production, and edible insects, have been proposed to overcome this shortage. Interestingly, edible insects are characterized by superior digestive and absorptive qualities that make them the ideal replacement for traditional protein production. This study aims to further the processing ability of insect protein by investigating the effects of various pre-treatment methods, such as blanching (HB), roasting (HR), and superheated steam (HS), on the nutritional properties and physicochemical characteristics of proteins extracted from Hermetia illucens larvae. The drying rate, pH value, color analysis, amino and fatty acid profile, as well as bulk density, shear force, and rehydration ratios of the above pre-treatment methods, were explored. HS was found to have the highest drying rate and pH value analysis showed that HB and HS samples have significantly higher values compared to the other modalities. Raw edible insects had the highest value in the sum of essential amino acid (EAA) and EAA index when compared to EAAs. HB and HS showed significantly lower bulk density results, and HS showed the highest shear force and the highest value in rehydration ratio, regardless of immersion time. Therefore, taking the above results together, it was found that blanching and superheated steam blanching pre-treatment were the most effective methods to improve the processing properties of H. illucens after hot-air drying.

Drying Techniques and Nutritional Composition of Drone Pupae (Apis mellifera L.) as Edible Food

  • Choi, Hong Min;Kim, Hyo-Young;Woo, Soon Ok;Kim, Se Gun;Bang, Kyeong Won;Moon, Hyo Jung;Han, Sang Mi
    • Journal of Apiculture
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    • v.34 no.2
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    • pp.161-167
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    • 2019
  • There is an urgent need for novel protein sources as an alternative to meat production. Insects, such as honeybees, hold potential as a safe, nutritious and reliable protein source for the future. In the present study, we established optimal powder preprocessing conditions of drone pupae (Apis mellifera L.) for use as a novel food. The content of moisture, crude protein, crude fat, crude ash, carbohydrate and crude fiber in drone pupae(Apis mellifera L.) were analyzed. The crude protein content ranged from 48.5 to 51.8% was found in both freeze-dried and hot-air powdered drone pupae. However, the protein content in the freeze-dried powder was higher than that in the hot-air powder by 3.3%. According to the Korean Food Standard Codex test method, coliforms, Salmonella spp. Staphylcoccus aureus, and Enterohamorrhagice Escherichia coli were not detected in both freeze-dried and hot-air powder. Therefore, we suggest that the high protein content of the powdered drone pupae prepared in this study can serve as a novel food.

Refolding of Acid-Unfolded Globin to Hemoglobin

  • Lee Jong-Woo
    • Biomedical Science Letters
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    • v.11 no.1
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    • pp.85-88
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    • 2005
  • Hemoglobin is oxygen carrier protein within erythrocyte in blood. Apoprotein of this, globin, is synthesized in the cytosol but it's cofactor, heme, is synthesized in the mitochondria. It has not been known very well how globin receives the heme from mitochondria and folds to hemoglobin. In this folding process, the initial structure of globin seems to be very important. A small volume of globin at acid pH was added rapidly into the bulk of an egg phosphatidylcholine $60\%$ liposome, containing hemins, at neutral pH according to the Rapid Dilution method. It was observed that an acid-induced unfolding structure of globin is initially needed to receive hemins from the lipid bilayer of liposomes. Also, this conclusion was confirmed with the absorption spectrum of the refolded globin separated by centrifugation.

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Cloning, Characterization, and Production of a Novel Lysozyme by Different Expression Hosts

  • Zhang, Haifeng;Fu, Gang;Zhang, Dawei
    • Journal of Microbiology and Biotechnology
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    • v.24 no.10
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    • pp.1405-1412
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    • 2014
  • Lysozyme is a protein found in egg white, tears, saliva, and other secretions. As a marketable natural alternative to preservatives, lysozyme can act as a natural antibiotic. In this study, we have isolated Bacillus licheniformis TIB320 from soil, which contains a lysozyme gene with various features. We have cloned and expressed the lysozyme in E. coli. The antimicrobial activity of the lysozyme showed that it had a broad antimicrobial spectrum against several standard strains. The lysozyme could maintain efficient activities in a pH range between 3 and 9 and from $20^{\circ}C$ to $60^{\circ}C$, respectively. The lysozyme was resistant to pepsin and trypsin to some extent at $40^{\circ}C$. Production of the lysozyme was optimized by using various expression strategies in B. subtilis WB800. The lysozyme from B. licheniformis TIB320 will be promising as a food or feed additive.

Preparation and Cytotoxicity Comparison of Type A Gelatin Nanoparticles with Recombinant Human Gelatin Nanoparticles

  • Won, Young-Wook;Kim, Yong-Hee
    • Macromolecular Research
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    • v.17 no.7
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    • pp.464-468
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    • 2009
  • Gelatin nanoparticles derived from bovine or porcine have been developed as various types of drug delivery system, and they need to be cross-linked to maintain their physicochemical properties in aqueous environments. Although gelatin is a widely used material in pharmaceutical industries, the safety issue of animal-origin gelatins, such as transmissible mad cow disease and anaphylaxis, remains to be solved. The purpose of this study was to prepare type A gelatin (GA) nanoparticles by modified, two-step, desolvation method and compare the toxicity of the resulting GA nanoparticles with recombinant human gelatin (rHG) nanoparticles. The GA nanoparticles were characterized, and drug loading and release pattern were measured. FITC-BSA, a model protein, was efficiently loaded in the nanoparticles and then released in a biphasic and sustained release pattern without an initial burst. In particular, the cell viability of the GA nanoparticles was less than that of the rHG nanoparticles. This finding suggests that rHG nanoparticles should be considered as an alternative to animal-origin gelatin nanoparticles in order to minimize the safety problems.

Molecular Basis of Drug Resistance: Epidermal Growth Factor Receptor Tyrosine Kinase Inhibitors and Anaplastic Lymphoma Kinase Inhibitors

  • Yang, Sei-Hoon
    • Tuberculosis and Respiratory Diseases
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    • v.75 no.5
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    • pp.188-198
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    • 2013
  • Over the past decade, several kinase inhibitors have been approved based on their clinical benefit in cancer patients. Unfortunately, in many cases, patients develop resistance to these agents via secondary mutations and alternative mechanisms. To date, several major mechanisms of acquired resistance, such as secondary mutation of the epidermal growth factor receptor (EGFR) gene, amplification of the MET gene and overexpression of hepatocyte growth factor, have been reported. This review describes the recent findings on the mechanisms of primary and acquired resistance to EGFR tyrosine kinase inhibitors and acquired resistance to anaplastic lymphoma kinase inhibitors, primarily focusing on non-small cell lung carcinoma.

Purification and Characterization of Crystalins by Aqueous Two-Phase Extraction

  • Bermudez, Ondrea;Forciniti, Daniel
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.6 no.6
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    • pp.395-401
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    • 2001
  • Crystallins are a family of water-soluble proteins that constitute up to 90% of the wa-ter-soluble proteins in mammalian eye lenses, We present in this paper an alternative purification method for these proteins using polyethylene glycol/dextran aqueous two-phase extraction. Un-der the appropriate conditions, we were able to recover the γ-crystallin fraction essentially free of the remaining proteins. High concentrations of salt at a neutral pH maximize the recovery of γ-crystallins in the top phase and minimize the contamination by the other proteins present in the lenses. The proposed protocol decreases the separation time by about 50%. The complex partition behavior observed for these proteins reflects a delicate balance between protein/phase-forming species(various polymers and salts) and protein interactions. This is evidenced, in part, by the role played by the largest proteins in this group as a "pseudo"phase-forming species.

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Farnesyl Protein Transferase Inhibitory Components of Lithospermum erythrorhizon

  • Kim, Seong-Jin;Kwon, Byoung-Mog;Kim, Sung-Hoon;Baek, Nam-In;Yang, Jae-Heon;Lee, Jeong-Joo;Lee, Sa-Im;Kwon, Young-Ee;Park, Hee-Wook;Lee, Jae-Hyeok;Park, Jeong-Suk;Kim, Dae-Keun
    • Natural Product Sciences
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    • v.13 no.4
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    • pp.328-331
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    • 2007
  • The methanolic extract of the roots of Lithospermum erythrorhizon (Boraginaceae) was found to show inhibitory activity towards farnesyl protein transferase (FPTase). Bioassay-guided fractionation of the methanolic extract resulted in the isolation of three naphthoquinone compounds, as inhibitors of FPTase. These compounds inhibited the FPTase activity in a dose-dependent manner.