Please use this identifier to cite or link to this item:
標題: The optimum production of recombinant type I antifreeze protein analogue by Bacillus subtilis and application on probiotic and cell cryopreservation
作者: 詹雅婷
Chan, Ya-Ting
關鍵字: Recombinant type I AFP (rAFP);重組第一型抗凍蛋白質類似物;Bacillus subtilis;cryopreservation;C2BBel cell;Lactobacillus;枯草桿菌;冷凍保護;細胞;益生菌
出版社: 食品暨應用生物科技學系所
引用: 王志鵬, 2001。一、枯草桿菌Bacillus subtilis DB104電轉型效率之增進。二、設計、合成第一型抗凍蛋白基因及其在枯草桿菌、大腸桿菌中之表現。國立中興大學食品科學系碩士論文。 蘇芳仙, 2001。最佳σA啟動子及多重啟動子之構築及其於枯草桿菌中之表現。國立中興大學食品科學系碩士論文。 戴君如, 2003。Bacillus subtilis 群之分類近況。食品工業。35(7):42-53 Guei-Cheng Peng, Ching-Hsiang Hsu,2005。熱滅活副乾酪乳酸桿菌治療屋塵蟎致慢性過敏性鼻炎的有效性和安全性。PEDIATRIC ALLERGY AND IMMUNOLOGY. 王志鵬, 2007。開發枯草桿菌持續型及誘導型表現系統以應用於自體、同源及異源蛋白質之表現暨建立芽孢桿菌益生菌表現系統。國立中興大學食品暨應用生物科技學系博士論文。 高苾瑜,2008。第一型重組抗凍蛋白質類似物之生產與其於食品工業之應用。國立中興大學食品暨應用生物科技學系碩士論文。 Akita, M., Sasaki, S., Matsuyama, S., & Mizushima, S. (1990). SecA interacts with secretory proteins by recognizing the positive charge at the amino terminus of the signal peptide in Escherichia coli. J Biol Chem, 265(14), 8164-8169. Amir, G., Rubinsky, B., Kassif, Y., Horowitz, L., Smolinsky, A. K., & Lavee, J. (2003). Preservation of myocyte structure and mitochondrial integrity in subzero cryopreservation of mammalian hearts for transplantation using antifreeze proteins--an electron microscopy study. Eur J Cardiothorac Surg, 24(2), 292-296; discussion 296-297. Briggs, M. S., & Gierasch, L. M. (1986). Molecular mechanisms of protein secretion: the role of the signal sequence. Adv Protein Chem, 38, 109-180. Carpenter, J. F., & Hansen, T. N. (1992). Antifreeze protein modulates cell survival during cryopreservation: mediation through influence on ice crystal growth. Proc Natl Acad Sci U S A, 89(19), 8953-8957. Chakrabartty ., Hew C. L., M., S., & L., F. G. (1988). Primary structures of the alanine rich antifreeze polypeptides from grubby sculpin (Myoxocephalus aenaeus). Can. J. Zool., 66, 403-408. Chao, H., Hodges, R. S., Kay, C. M., Gauthier, S. Y., & Davies, P. L. (1996). A natural variant of type I antifreeze protein with four ice-binding repeats is a particularly potent antifreeze. Protein Sci, 5(6), 1150-1156. Cutler, A. J. (1989). Winter flounder antifreeze protein improves the cold hardiness of plant tissues. Journal of Plant Physiology, 135(3), 351. Davies, P. L., & Hew, C. L. (1990). Biochemistry of fish antifreeze proteins. FASEB J, 4(8), 2460-2468. Davies, P. L., Roach, A. H., & Hew, C. L. (1982). DNA sequence coding for an antifreeze protein precursor from winter flounder. Proc Natl Acad Sci U S A, 79(2), 335-339. Deng, G., Andrews, D. W., & Laursen, R. A. (1997). Amino acid sequence of a new type of antifreeze protein, from the longhorn sculpin Myoxocephalus octodecimspinosis. FEBS Lett, 402(1), 17-20. Deuerling, E., Mogk, A., Richter, C., Purucker, M., & Schumann, W. (1997). The ftsH gene of Bacillus subtilis is involved in major cellular processes such as sporulation, stress adaptation and secretion. Mol Microbiol, 23(5), 921-933. DeVries, A. L., & Wohlschlag, D. E. (1969). Freezing resistance in some Antarctic fishes. Science, 163(871), 1073-1075. Donald, G. (1994). Actomyosin Stabilization to Freeze‐Thaw and Heat Denaturation by Lactate Salts. Journal of Food Science, 59(1), 101. Doucet, C. J., Byass, L., Elias, L., Worrall, D., Smallwood, M., & Bowles, D. J. (2000). Distribution and characterization of recrystallization inhibitor activity in plant and lichen species from the UK and maritime Antarctic. Cryobiology, 40(3), 218-227. Duman, J. G. (1979). Subzero temperature tolerance in spiders: The role of thermal-hysteresis-factors. Journal of Comparative Physiology A-Neuroethology Sensory neural And Behavioral Physiology, 131(4), 347. Duman, J. G. (2004). Antifreeze proteins in Alaskan insects and spiders. Journal of Insect Physiology, 50(4), 259. Duman, J. G., Li, N., Verleye, D., Goetz, F. W., Wu, D. W., Andorfer, C. A., Benjamin, T., & Parmelee, D. C. (1998). Molecular characterization and sequencing of antifreeze proteins from larvae of the beetle Dendroides canadensis. J Comp Physiol B, 168(3), 225-232. Duman, J. G., & Olsen, T. M. (1993). Thermal Hysteresis Protein Activity in Bacteria, Fungi, and Phylogenetically Diverse Plants. Cryobiology, 30(3), 322-328. Ewart, K. V., & Fletcher, G. L. (1993). Herring antifreeze protein: primary structure and evidence for a C-type lectin evolutionary origin. Mol Mar Biol Biotechnol, 2(1), 20-27. Ewart, K. V., Lin, Q., & Hew, C. L. (1999). Structure, function and evolution of antifreeze proteins. Cell Mol Life Sci, 55(2), 271-283. Ewart, K. V., Rubinsky, B., & Fletcher, G. L. (1992). Structural and functional similarity between fish antifreeze proteins and calcium-dependent lectins. Biochem Biophys Res Commun, 185(1), 335-340. Ewart, K. V., Yang, D. S., Ananthanarayanan, V. S., Fletcher, G. L., & Hew, C. L. (1996). Ca2+-dependent antifreeze proteins. Modulation of conformation and activity by divalent metal ions. J Biol Chem, 271(28), 16627-16632. Feeney, R. E., Burcham, T. S., & Yeh, Y. (1986). Antifreeze glycoproteins from polar fish blood. Annu Rev Biophys Biophys Chem, 15, 59-78. Feeney, R. E., & Yeh, Y. (1978). Antifreeze proteins from fish bloods. Adv Protein Chem, 32, 191-282. Graham, L. A., Liou, Y. C., Walker, V. K., & Davies, P. L. (1997). Hyperactive antifreeze protein from beetles. Nature, 388(6644), 727-728. Griffith, M., Ala, P., Yang, D. S., Hon, W. C., & Moffatt, B. A. (1992). Antifreeze protein produced endogenously in winter rye leaves. Plant Physiol, 100(2), 593-596. Griffith, M., & Ewart, K. V. (1995). Antifreeze proteins and their potential use in frozen foods. Biotechnol Adv, 13(3), 375-402. Harding, M. M., Ward, L. G., & Haymet, A. D. (1999). Type I ''antifreeze'' proteins. Structure-activity studies and mechanisms of ice growth inhibition. Eur J Biochem, 264(3), 653-665. Hew, C. L. (1983). Presence of cystine-containing antifreeze proteins in the spruce bud worm, Choristoneura fumiferana. Canadian Journal of Zoology, 61(10), 2324. Hew, C. L., Joshi, S., Wang, N. C., Kao, M. H., & Ananthanarayanan, V. S. (1985). Structures of shorthorn sculpin antifreeze polypeptides. Eur J Biochem, 151(1), 167-172. Hon, W. C., Griffith, M., Mlynarz, A., Kwok, Y. C., & Yang, D. S. (1995). Antifreeze proteins in winter rye are similar to pathogenesis-related proteins. Plant Physiol, 109(3), 879-889. Huang, T., & Duman, J. G. (2002). Cloning and characterization of a thermal hysteresis (antifreeze) protein with DNA-binding activity from winter bittersweet nightshade, Solanum dulcamara. Plant Mol Biol, 48(4), 339-350. Husby, J. A. (1980). Antifreeze agents in the body fluid of winter active insects and spiders. cellular and molecular life sciences, 36(8), 963. In ''t Veld, G., Driessen, A. J., & Konings, W. N. (1992). Effect of the unsaturation of phospholipid acyl chains on leucine transport of Lactococcus lactis and membrane permeability. Biochim Biophys Acta, 1108(1), 31-39. Jia, Z., DeLuca, C. I., Chao, H., & Davies, P. L. (1996). Structural basis for the binding of a globular antifreeze protein to ice. Nature, 384(6606), 285-288. Kenward, K. D., Altschuler, M., Hildebrand, D., & Davies, P. L. (1993). Accumulation of type I fish antifreeze protein in transgenic tobacco is cold-specific. Plant Mol Biol, 23(2), 377-385. Kets, E., Teunissen, P., & de Bont, J. (1996). Effect of compatible solutes on survival of lactic Acid bacteria subjected to drying. Appl Environ Microbiol, 62(1), 259-261. Kristiansen, E., Ramlov, H., Hagen, L., Pedersen, S. A., Andersen, R. A., & Zachariassen, K. E. (2005). Isolation and characterization of hemolymph antifreeze proteins from larvae of the longhorn beetle Rhagium inquisitor (L.). Comp Biochem Physiol B Biochem Mol Biol, 142(1), 90-97. Li, N., Chibber, B. A., Castellino, F. J., & Duman, J. G. (1998). Mapping of disulfide bridges in antifreeze proteins from overwintering larvae of the beetle Dendroides canadensis. Biochemistry, 37(18), 6343-6350. Li, X. M., Trinh, K. Y., Hew, C. L., Buettner, B., Baenziger, J., & Davies, P. L. (1985). Structure of an antifreeze polypeptide and its precursor from the ocean pout, Macrozoarces americanus. J Biol Chem, 260(24), 12904-12909. Lin, F., Zi Rong, X., Wei Fen, L., Jiang Bing, S., Ping, L., & Chun Xia, H. (2007). Protein secretion pathways in Bacillus subtilis: implication for optimization of heterologous protein secretion. Biotechnol Adv, 25(1), 1-12. Liou, Y. C., Thibault, P., Walker, V. K., Davies, P. L., & Graham, L. A. (1999). A complex family of highly heterogeneous and internally repetitive hyperactive antifreeze proteins from the beetle Tenebrio molitor. Biochemistry, 38(35), 11415-11424. Lu, C. F. (2000). Secretory antifreeze proteins produced in suspension culture cells of Rhodiola algida var. tangutica during cold acclimation. Progress in Biochemistry and Biophysics, 27(5), 555. Lubzens, E., Daube, N., Pekarsky, I., Magnus, Y., Cohen, A., Yusefovich, F., & Feigin, P. (1997). Carp (Cyprinus carpio L.) spermatozoa cryobanks -- strategies in research and application. Aquaculture, 155(1-4), 13-30. Meyer, K., Keil, M., & Naldrett, M. J. (1999). A leucine-rich repeat protein of carrot that exhibits antifreeze activity. FEBS Lett, 447(2-3), 171-178. Miles, A. A., Misra, S. S., & Irwin, J. O. (1938). The estimation of the bactericidal power of the blood. J Hyg (Lond), 38(6), 732-749. Miller, J. F. (1994). Bacterial transformation by electroporation. Methods in Enzymology, 235, 375. Mirel, D. B., Estacio, W. F., Mathieu, M., Olmsted, E., Ramirez, J., & Marquez-Magana, L. M. (2000). Environmental regulation of Bacillus subtilis sigma(D)-dependent gene expression. J Bacteriol, 182(11), 3055-3062. Mueller, G. M., McKown, R. L., Corotto, L. V., Hague, C., & Warren, G. J. (1991). Inhibition of recrystallization in ice by chimeric proteins containing antifreeze domains. J Biol Chem, 266(12), 7339-7344. Ng, N. F., & Hew, C. L. (1992). Structure of an antifreeze polypeptide from the sea raven. Disulfide bonds and similarity to lectin-binding proteins. J Biol Chem, 267(23), 16069-16075. Ng, N. F., Trinh, K. Y., & Hew, C. L. (1986). Structure of an antifreeze polypeptide precursor from the sea raven, Hemitripterus americanus. J Biol Chem, 261(33), 15690-15695. Panoff, J. M., Thammavongs, B., Gueguen, M., & Boutibonnes, P. (1998). Cold stress responses in mesophilic bacteria. Cryobiology, 36(2), 75-83. Payne, S. R., & Young, O. A. (1995). Effects of pre-slaughter administration of antifreeze proteins on frozen meat quality. Meat Science, 41(2), 147-155. Robles, V., Cabrita, E., Anel, L., & Herráez, M. P. (2006). Microinjection of the antifreeze protein type III (AFPIII) in turbot (Scophthalmus maximus) embryos: Toxicity and protein distribution. Aquaculture, 261(4), 1299-1306. Scott, Davies P. L., A., S. M., & L., F. G. (1987). Structural variation in the alanine-rich antifreeze proteins of the Pleuronectidae. Eur. J. Biochem., 168, 629-633. Scott, G. K., Hayes, P. H., Fletcher, G. L., & Davies, P. L. (1988). Wolffish antifreeze protein genes are primarily organized as tandem repeats that each contain two genes in inverted orientation. Mol Cell Biol, 8(9), 3670-3675. Sicheri, F., & Yang, D. S. (1995). Ice-binding structure and mechanism of an antifreeze protein from winter flounder. Nature, 375(6530), 427-431. Sinclair, B. J. (2002). Haemolymph osmolality and thermal hysteresis activity in 17 species of arthropods from sub-Antarctic Marion Island. POLAR BIOLOGY, 25(12), 928. Smallwood, M., Worrall, D., Byass, L., Elias, L., Ashford, D., Doucet, C. J., Holt, C., Telford, J., Lillford, P., & Bowles, D. J. (1999). Isolation and characterization of a novel antifreeze protein from carrot (Daucus carota). Biochem J, 340 ( Pt 2), 385-391. Su-Mei Wu, P.-P. H., Choy-Leong Hew and Jen-Leih Wu (1998). Effect of antifreeze protein on cold tolerance in juvenile tilapia (Oreochromis mossambicus Peters) and milkfish (Chanos chanos Forsskal). Zoological Studies, 37, 39-44. Tamiya, T. (1985). Freeze denaturation of enzymes and its prevention with additives. Cryobiology, 22(5), 446. Tjalsma, H., Antelmann, H., Jongbloed, J. D., Braun, P. G., Darmon, E., Dorenbos, R., Dubois, J. Y., Westers, H., Zanen, G., Quax, W. J., Kuipers, O. P., Bron, S., Hecker, M., & van Dijl, J. M. (2004). Proteomics of protein secretion by Bacillus subtilis: separating the "secrets" of the secretome. Microbiol Mol Biol Rev, 68(2), 207-233. Tjalsma, H., Bolhuis, A., Jongbloed, J. D., Bron, S., & van Dijl, J. M. (2000). Signal peptide-dependent protein transport in Bacillus subtilis: a genome-based survey of the secretome. Microbiol Mol Biol Rev, 64(3), 515-547. Tursman, D. (1994). Freeze tolerance adaptations in the centipede, Lithobius forficatus. Journal of Experimental Zoology Part A: Ecological Genetics and Physiology, 268(5), 347. Tursman, D. (1995). Cryoprotective effects of thermal hysteresis protein on survivorship of frozen gut cells from the freeze-tolerant centipede Lithobius forficatus. Journal of Experimental Zoology Part A: Ecological Genetics and Physiology, 272(4), 249. Tyshenko, M. G., Doucet, D., Davies, P. L., & Walker, V. K. (1997). The antifreeze potential of the spruce budworm thermal hysteresis protein. Nat Biotechnol, 15(9), 887-890. Urrutia, M. E., Duman, J. G., & Knight, C. A. (1992). Plant thermal hysteresis proteins. Biochim Biophys Acta, 1121(1-2), 199-206. von Heijne, G. (1985). Signal sequences. The limits of variation. J Mol Biol, 184(1), 99-105. Waites, W. M., Kay, D., Dawes, I. W., Wood, D. A., Warren, S. C., & Mandelstam, J. (1970). Sporulation in Bacillus subtilis. Correlation of biochemical events with morphological changes in asporogenous mutants. Biochem J, 118(4), 667-676. Wallis, J. G., Wang, H., & Guerra, D. J. (1997). Expression of a synthetic antifreeze protein in potato reduces electrolyte release at freezing temperatures. Plant Mol Biol, 35(3), 323-330. Wang, J. P., Yeh, C. M., & Tsai, Y. C. (2006). Improved subtilisin YaB production in Bacillus subtilis using engineered synthetic expression control sequences. J Agric Food Chem, 54(25), 9405-9410. Wang, R., Zhang, P., Gong, Z., & Hew, C. L. (1995). Expression of the antifreeze protein gene in transgenic goldfish (Carassius auratus) and its implication in cold adaptation. Mol Mar Biol Biotechnol, 4(1), 20-26. Warren, G. J., Mueller, G. M., & McKown, R. L. (1992). Ice crystal growth suppression polypeptides and method of making. U.S. Patent 5118792. Wen, D., & Laursen, R. A. (1992). Structure-function relationships in an antifreeze polypeptide. The role of neutral, polar amino acids. J Biol Chem, 267(20), 14102-14108. Wierzbicki, A., Dalal, P., Cheatham, T. E., 3rd, Knickelbein, J. E., Haymet, A. D., & Madura, J. D. (2007). Antifreeze proteins at the ice/water interface: three calculated discriminating properties for orientation of type I proteins. Biophys J, 93(5), 1442-1451. Wolfe, J., & Bryant, G. (1999). Freezing, drying, and/or vitrification of membrane- solute-water systems. Cryobiology, 39(2), 103-129. Yang, C., & Sharp, K. A. (2005). Hydrophobic tendency of polar group hydration as a major force in type I antifreeze protein recognition. Proteins, 59(2), 266-274. Yeh, C. M., Wang, J. P., Lo, S. C., Chan, W. C., & Lin, M. Y. (2010). Chromosomal integration of a synthetic expression control sequence achieves poly-gamma-glutamate production in a Bacillus subtilis strain. Biotechnol Prog, 26(4), 1001-1007. Yeh, Y., & Feeney, R. E. (1996). Antifreeze Proteins: Structures and Mechanisms of Function. Chem Rev, 96(2), 601-618. Younis, A. I., Rooks, B., Khan, S., & Gould, K. G. (1998). The effects of antifreeze peptide III (AFP) and insulin transferrin selenium (ITS) on cryopreservation of chimpanzee (Pan troglodytes) spermatozoa. J Androl, 19(2), 207-214. Yu, X. M., & Griffith, M. (2001a). Winter rye antifreeze activity increases in response to cold and drought, but not abscisic acid. Physiol Plant, 112(1), 78-86. Yu, X. M., Griffith, M., & Wiseman, S. B. (2001b). Ethylene induces antifreeze activity in winter rye leaves. Plant Physiol, 126(3), 1232-1240.
抗凍蛋白質(antifreeze proteins, AFP)存在於某些寒帶地區生物如魚類、植物和細菌體內,具有修飾冰晶構形,抑制冰晶生長及抑制冰晶再結晶之功能。可應用於冷凍食品之品質保存或細胞、精卵之冷凍保存及非耐寒生物轉殖抗凍蛋白基因降低寒害等。
枯草桿菌(Bacillus subtilis)為GRAS (generally recognized as safe)級之菌種,常被運用於發酵工業。此外在遺傳工程研究範圍中,亦常被使用做為宿主細胞表現異源蛋白質。本實驗室先前已成功建立枯草桿菌分泌表現重組第一型抗凍蛋白質類似物之系統,但產量仍有待提升。
本研究建立枯草桿菌持續型和誘導型表現系統表現重組第一型抗凍蛋白質類似物,其中以B. subtilis(pSECS-6AFPT) 發酵可得最高產量為872 mg/L。將上清液純化所得之重組第一型抗凍蛋白質類似物經加熱1.5小時或改變緩衝液pH值範圍於2~12後仍具有抗凍活性。重組第一型抗凍蛋白類似物應用方面,益生菌冷凍保護部分,重組第一型抗凍蛋白類似物搭配海藻糖,脫脂奶粉等保護劑可提高Lactobacillus paracasei菌體的冷凍儲存存活率。於菌體凍乾實驗中,在適當儲藏條件下,重組第一型抗凍蛋白類似物可略為提升L. paracasei存活率。細胞冷凍保護部分,重組第一型抗凍蛋白類似物雖無顯著差異但可略為提升DMSO保護劑處理之C2BBel細胞冷凍存活率。本實驗室研發之重組第一型抗凍蛋白質未來可嘗試應用於其他生物素材之冷凍保護,增加其應用潛力。

Antifreeze proteins (AFPs) play an important role in the freezing tolerance of various organisms such as fish, plants, and bacteria who live in supercooling conditions. It modifies the shape of ice crystal, inhibits ice growth, and represses the ice crystals from recrystallization. AFPs have potential to apply in food technology, cryopreservation, cryosurgery and transgenic technologies.
Bacillus subtilis has been classified as GRAS (generally recognized as safe) microorganism by FDA and considered as an excellent host for the secretory production of heterologous proteins.In our previous study, Synthetic gene encoding the recombinant type I AFP (rAFP) analogue has been expressed in Bacillus subtilis, but the production yield needed further improvement.
In this study, constitutive and inducible system of B. subtilis were constructed to optimize the rAFP production yield. B. subtilis (pSECS-6AFPT) had the maximum yield of 872 mg/L. The rAFP heated for 1.5 hr or changed in various pH buffers ranging from 2-12 still retained their antifreeze activity.
The application of rAFP in probiotic cryopreservation showed that rAFP together with other cryoprotectants such as trehalose or skim milk improved the viability of frozen Lactobacillus paracasei. In proper storage condition,rAFP slightly improved the viability of lyophilized L. paracasei. In C2BBel cell cryopreservation experiments, rAFP slightly but not significantly improve the DMSO treated frozen C2BBel cells viability. The potent application of rAFP on biological materials cryopreservation is highly expected in the future.
其他識別: U0005-2607201116174000
Appears in Collections:食品暨應用生物科技學系

Show full item record

Google ScholarTM


Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.