• Title/Summary/Keyword: Alternative protein

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Heterologous Expression of Interferon α-2b in Lactococcus lactis and its Biological Activity against Colorectal Cancer Cells

  • Meilina, Lita;Budiarti, Sri;Mustopa, Apon Zaenal;Darusman, Huda Shalahudin;Triratna, Lita;Nugraha, Muhammad Ajietuta;Bilhaq, Muhammad Sabiq;Ningrum, Ratih Asmana
    • Microbiology and Biotechnology Letters
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    • v.49 no.1
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    • pp.75-87
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    • 2021
  • Type I Interferons (IFNα) are known for their role as biological anticancer agents owing to their cell-apoptosis inducing properties. Development of an appropriate, cost-effective host expression system is crucial for meeting the increasing demand for proteins. Therefore, this study aims to develop codon-optimized IFNα-2b in L. lactis NZ3900. These cells express extracellular protein using the NICE system and Usp45 signal peptide. To validate the mature form of the expressed protein, the recombinant IFNα-2b was screened in a human colorectal cancer cell line using the cytotoxicity assay. The IFNα-2b was successfully cloned into the pNZ8148 vector, thereby generating recombinant L. lactis pNZ8148-SPUsp45-IFNα-2b. The computational analysis of codon-optimized IFNα-2b revealed no mutation and amino acid changes; additionally, the codon-optimized IFNα-2b showed 100% similarity with native human IFNα-2b, in the BLAST analysis. The partial size exclusion chromatography (SEC) of extracellular protein yielded a 19 kDa protein, which was further confirmed by its positive binding to anti-IFNα-2b in the western blot analysis. The crude protein and SEC-purified partial fraction showed IC50 values of 33.22 ㎍/ml and 127.2 ㎍/ml, respectively, which indicated better activity than the metabolites of L. lactis NZ3900 (231.8 ㎍/ml). These values were also comparable with those of the regular anticancer drug tamoxifen (105.5 ㎍/ml). These results demonstrated L. lactis as a promising host system that functions by utilizing the pNZ8148 NICE system. Meanwhile, codon-optimized usage of the inserted gene increased the optimal protein expression levels, which could be beneficial for its large-scale production. Taken together, the recombinant L. lactis IFNα-2b is a potential alternative treatment for colorectal cancer. Furthermore, its activity was analyzed in the WiDr cell line, to assess its colorectal anticancer activities in vivo.

Microscopic Feature, Protein Marker Expression, and Osteoinductivity of Human Demineralized Dentin Matrix

  • Park, Sung-Min;Hwang, Jung-Kook;Kim, Young-Kyun;Um, In-Woong;Lee, Geun-Ho;Kim, Kyung-Wook
    • Journal of Korean Dental Science
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    • v.5 no.2
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    • pp.77-87
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    • 2012
  • Purpose: This study examined the scanning electron microscopic feature, protein marker expression and osteoinductive activity of demineralized dentin matrix (DDM) from human for nude mice. Materials and Methods: Twenty healthy nude mice, weighing about 20 g were used for study. DDM from Human was prepared and implanted into the dorsal portion of nude mouse. Before implantation, DDM was examined by scanning electron microscopy (SEM). Nude mice were sacrificed at 2 weeks, 4 weeks and 8 weeks after DDM grafting and evaluated histologically by H-E, MT staining. And also immunohistochemistry analysis (ostecalcin, osteopontin) was performed. Result: Dentinal tubules and collagen fibers were observed by SEM of dentin surface of DDM. The DDM induced bone and cartilage independently in soft tissues. And, the histological findings showed bone forming cells like osteoblasts, fibroblasts at 2, 4 and 8 weeks. On immunohistochemistry analysis, osteocalcin and osteopontin positive bone forming cells were observed. Conclusion: This results showed that the DDM from human has osteoinductive ability and is a good alternative to autogenous bone graft materials.

Natural language processing techniques for bioinformatics

  • Tsujii, Jun-ichi
    • Proceedings of the Korean Society for Bioinformatics Conference
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    • 2003.10a
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    • pp.3-3
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    • 2003
  • With biomedical literature expanding so rapidly, there is an urgent need to discover and organize knowledge extracted from texts. Although factual databases contain crucial information the overwhelming amount of new knowledge remains in textual form (e.g. MEDLINE). In addition, new terms are constantly coined as the relationships linking new genes, drugs, proteins etc. As the size of biomedical literature is expanding, more systems are applying a variety of methods to automate the process of knowledge acquisition and management. In my talk, I focus on the project, GENIA, of our group at the University of Tokyo, the objective of which is to construct an information extraction system of protein - protein interaction from abstracts of MEDLINE. The talk includes (1) Techniques we use fDr named entity recognition (1-a) SOHMM (Self-organized HMM) (1-b) Maximum Entropy Model (1-c) Lexicon-based Recognizer (2) Treatment of term variants and acronym finders (3) Event extraction using a full parser (4) Linguistic resources for text mining (GENIA corpus) (4-a) Semantic Tags (4-b) Structural Annotations (4-c) Co-reference tags (4-d) GENIA ontology I will also talk about possible extension of our work that links the findings of molecular biology with clinical findings, and claim that textual based or conceptual based biology would be a viable alternative to system biology that tends to emphasize the role of simulation models in bioinformatics.

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Molecular Cloning, Identification and Characteristics of a Novel Isoform of Carbamyl Phosphate Synthetase I in Human Testis

  • Huo, Ran;Zhu, Hui;Lu, Li;Ying, Lanlan;Xu, Min;Xu, Zhiyang;Li, Jianmin;Zhou, Zuomin;Sha, Jiahao
    • BMB Reports
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    • v.38 no.1
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    • pp.28-33
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    • 2005
  • A gene coding a novel isoform of carbamyl phosphate synthetase I (CPS1) was cloned from a human testicular library. As shown by cDNA microarray hybridization, this gene was expressed at a higher level in human adult testes than in fetal testes. The full length of its cDNA was 3831 bp, with a 3149 bp open reading frame, encoding a 1050-amino-acid protein. The cDNA sequence was deposited in the GenBank (AY317138). Sequence analysis showed that it was homologous to the human CPS1 gene. The putative protein contained functional domains composing the intact large subunit of carbamoyl phosphate synthetase, thus indicated it has the capability of arginine biosynthesis. A multiple tissue expression profile showed high expression of this gene in human testis, suggesting the novel alternative splicing form of CPS1 may be correlated with human spermatogenesis.

Plant Toxins and Detoxification Methods to Improve Feed Quality of Tropical Seeds - Review -

  • Makkar, H.P.S.;Becker, K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.12 no.3
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    • pp.467-480
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    • 1999
  • Many antinutritional and toxic factors abound in tropical seeds, which are also generally rich in nutrients and therefore more prone to attack from herbivores. Antinutritional and toxic factors are considered to defend seeds against environmental vagaries and thus help to protect them. These factors though good for the plant, cause deleterious effects or are even toxic to animals and man. The conventional seeds cultivated for oil or non-oil purposes, and general aspects of antinutritional factors are not presented here as these have already been discussed widely by many workers. Deficits in conventional protein and energy sources in the tropics have stimulated a quest for alternative feeds both for animals and humans. This article attempts to highlight two new oilseed crops, Jatropha curcas and Moringa oleifera, and in addition deals with some under-utilized seeds with potential as animal feed. Most of these seed plants are adapted to various marginal growing conditions in the tropics and can help to mitigate the prevailing deficit in protein and energy sources. Antinutritional and toxic factors in seed or seed meal, various approaches to detoxify seed meal, and future research and development priorities for their exploitation as animal feeds are presented.

A Recent Research Trends for Food Emulsions using Pickering Stabilization of Nano-particles (나노 입자의 피커링 안정화를 이용한 식품 에멀젼의 최근 연구동향)

  • Cho, Wan-Goo
    • Journal of the Korean Applied Science and Technology
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    • v.29 no.2
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    • pp.238-247
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    • 2012
  • Nanotechnology in the food industry is an emerging area with considerable research and potential products. Solid particles of nanoscale and microscale dimensions are becoming recognized for their potential application in the formulation of novel dispersed systems containing emulsified oil or water droplets. This review describes developments in the formation and properties of food-grade emulsion systems based on edible fat crystals, silica nanoparticles, and novel particles of biological origin nanocrystals. The special features characterizing the properties of Pickering stabilized droplets are focused in comparison with those of protein-stabilized emulsions. We also review describes application examples of these in the food industry.

Simple Purification of the Human Antimicrobial Peptide Dermcidin (MDCD-1L) by Intein-Mediated Expression in E. coli

  • Hong, In-Pyo;Kim, Yong-Seok;Choi, Shin-Geon
    • Journal of Microbiology and Biotechnology
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    • v.20 no.2
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    • pp.350-355
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    • 2010
  • Among human antimicrobial peptides (hAMPs), DCD-1L has a broad spectrum of antimicrobial activity over a wide pH range and in high salt concentrations. It offers a promising alternative to conventional antibiotics. The 458-bp-long dermcidin cDNA was amplified by PCR using a human fetal cDNA library as a template. The 147-bp fragment of the MDCD-1L gene encoding an additional methionine residue was subcloned into the pTYB11 vector. Recombinant MDCD-1L was expressed as an intein fusion protein in E. coli, and then purified by affinity chromatography using chitin beads. A small peptide with a molecular mass of about 5 kDa was detected by tricine gel electrophoresis. The recombinant MDCD-1L peptide was purified from the gel and its amino acid sequence was determined by nanoLC-ESI-MS/MS analysis. The initiating amino acid, methionine, remained attached to the N-terminal region of recombinant MDCD-1L. Purified MDCD-1L showed antimicrobial activity against a Micrococcus luteus test strain.

Reconsideration of Classical Antibiotic Lincomycin: Anti-inflammatory Effect in LPS-stimulated RAW 264.7 Cells

  • Yang, Eun-Jin;Lee, Nari;Hyun, Chang-Gu
    • Microbiology and Biotechnology Letters
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    • v.48 no.3
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    • pp.366-372
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    • 2020
  • Since, side effects of antibiotics are frequently emphasized these days, their use is gradually diminishing, and alternative drugs are being developed. We have sought to reintroduce them as raw materials for human health as conventional 'weapons' that have been retired after their historical duties. In this study, we investigated the anti-inflammatory effects of lincomycin (LIN), on lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. Our findings show that LIN potently inhibited production of LPS-induced proinflammatory mediators, such as nitric oxide (NO) and prostaglandin E2 (PGE2), without cytotoxicity. Consistent with these findings, LIN strongly decreased protein expression levels of inducible NO synthase (iNOS) and cyclooxygenase (COX-2). Furthermore, LIN reduced pro-inflammatory cytokines such as tumor necrosis factor (TNF)-α, interleukin (IL)-6, and IL-1β. To further elucidate the mechanisms of these inhibitory effects of LIN, we studied LPS-induced IκB-α degradation, and mitogen-activated protein kinase (MAPK) phosphorylation. LIN suppressed downregulation of inhibitory κB (IκB-α) degradation, and the phosphorylation of the c-Jun N-terminal kinase (JNK) pathway. Based on these results, we suggest that LIN may be considered a potential candidate as an anti-inflammatory cosmetic or a medicine for human health.

Polymorphism Assessment of Six Lentil (Lens culinaris Medik.) Genotypes Using Isozyme

  • Madina, M. Hur;Rahman, M. Saifur;Deb, A. Chandra;Choi, Yun Hee;Kim, Mi Ri;Shin, Jihoon;Yoo, Jin Cheol
    • Journal of Integrative Natural Science
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    • v.8 no.2
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    • pp.117-127
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    • 2015
  • Lentil (Lens culinaris Medik.) is one of the important legumes and cheaper source of protein in Bangladesh that displays great biological diversity. Isozyme, one of the most important protein markers to detect genetic polymorphism in lentil, whereas we considered thirteen-isozyme in six varieties viz., BARI masur-1, BARI masur-2, BARI masur-3, BARI masur-4, BARI masur-5 and BARI masur-6. The highest polymorphism was found in tyrosinase isozyme system. UPGMA analysis revealed that the highest similarity between BARI masur-5 and BARI masur-6 whereas, the highest genetic distance between BARI masur-1 and BARI masur-5 reflecting higher intervarietal variation. Principal component analysis (PCA) also revealed the similar results that of unweighted pair group method with arithmetic mean (UPGMA). The first, second and third PCs contributed 81.58%, 11.19% and 4.94% variation respectively, with cumulative variation of the first three PCs was 75.45%. Consequently, Isozyme could clearly assed the genetic diversity at intervarietal levels and these two varieties can be considered as valuable gene resources for future breeding and conservation programs.

Optimum Condition of Extracting Collagen from Chicken Feet and its Characetristics

  • Liu, D.C.;Lin, Y.K.;Chen, M.T.
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.11
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    • pp.1638-1644
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    • 2001
  • The objective of this research was to evaluate alternative treatments for the best extraction condition for collagen from chicken feet. Various properties such as chemical composition, amino acid, pH, swelling percentage, yield and pure collagen, collagen loss, color (Hunter L, a and b) and electrophoresis of collagen from chicken feet treated by 5% acids (acetic acid, citric acid. hydrochloric acid and lactic acid) and soaking times (12, 24, 36 and 48 h) were evaluated. The crude protein, fat, ash and moisture contents of chicken feet was 17.42, 12.04, 5.98 and 62.05%, respectively. Amino acid composition of collagen from chicken feet indicated that the protein of collagen was markedly hydrolized by the hydrochloric acid treatment. The result of electrophoresis also supported this phenomenon. Both the swelling percentage of lactic acid and citric acid treatments were significantly higher than that of acetic acid and HC1 treatment. The pH of the acid treatments ranged from 2.43-3.62. According to the result of yield, pure collagen and loss of collagen, the best condition of extracting collagen from chicken feet was soaked in 5% lactic acid for 36 h. However, a brighter yellow color of collagen from all treatments was observed with a longer soaking time.