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研究生:張文智
研究生(外文):Wen-Chih Chang
論文名稱:利用Bacilluscereus發酵蝦蟹殼廢棄物所生產抗真菌酵素之分離純化及其應用
論文名稱(外文):The purification and application of antifungal enzymes produced from shrimp and crab shell wastes by Bacillus cereus
指導教授:陳錦樹陳錦樹引用關係
指導教授(外文):Chinshuh Chen
學位類別:博士
校院名稱:國立中興大學
系所名稱:食品科學系
學門:農業科學學門
學類:食品科學類
論文種類:學術論文
論文出版年:2003
畢業學年度:91
語文別:中文
中文關鍵詞:抗真菌蝦蟹殼粉幾丁質酶幾丁聚醣酶蛋白酶臍帶血單核細胞
外文關鍵詞:antifungalshrimp and crab shell powderchitinasechitosanaseproteaseumbilical cord bloodmononuclear cell
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利用水產加工廢棄物可生產較廉價的抗真菌之水解酵素,故本研究主要利用經過煮沸及破碎處理而成之蝦蟹殼粉,用來培養具有抑制真菌及幾丁質水解能力的菌株。Bacillus cereus YQ308 發酵蝦蟹殼粉為主的培養基,能分泌幾丁質酶、幾丁聚醣酶及蛋白酶等抗真菌之水解酵素。YQ308菌株之發酵液能明顯影響Fusarium oxysporum、 Fusarium solani及 Pythium ultimum等植物病原性真菌之孢子發芽及芽管延長。
利用色層分析法可純化出幾丁質酶及蛋白酶,以SDS-PAGE測得分子量分別為48和 29 kDa。幾丁質酶的最適pH為7,最適溫度為50°С,pH的安定性為6-8,而熱安定性為60°С。
Bacillus cereus YQ 308所純化的幾丁質酶發現具有較高的乙二醇幾丁質酶及少許的乙二醇幾丁聚醣酶活性,但不具有N-乙醯葡萄糖胺酶活性。而水解懸浮態幾丁質主要產物為N-乙醯幾丁四糖,其次是N-乙醯幾丁二糖,根據此酵素的基質特異性及其主要水解產物分析判斷,應為內切型幾丁質酶。純化之幾丁質酶能強烈抑制F. oxysporum、 F. solani 和 P. ultimum 生物量。並對此幾丁質酶進行基因選殖,作為往後將此基因轉殖至酵母系統,使其大量表現作為生物防治之依據。
N-乙醯幾丁六醣和幾丁六醣具有增強免疫功能,然而其製備成本過高,無法實際應用。利用YQ 308幾丁質酶水解所得之0.065 mg/ml N-乙醯幾丁寡醣混合液,免疫活性最佳,可刺激臍帶血中單核細胞的增生。
The production of inexpensive antifungal hydrolytic enzymes is an element in the utilization of shellfish processing wastes. In this study, shrimp and crab shell powder prepared by treating shrimp and crab processing wastes with boiling and crushing was used as a substrate for the culturing of an antifungal chitinase producing microorganism. Bacillus cereus YQ308 produced antifungal hydrolytic enzymes, including chitinase, chitosanase and protease, when it was grown in a medium containing shrimp and crab shell powder (SCSP) obtained from marine waste. The growth of the plant-pathogenic fungi Fusarium oxysporum, Fusarium solani, and Pythium ultimum were significantly affected in the presence of the YQ308 culture supernatant. The supernatant inhibited spore germination and germ tube elongation of F. oxysporum, F. solani, and P. ultimum.
The chitinase and protease, purified by sequential chromatography, had molecular weigfts of 48 and 29 kDa, respectively, by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The pI of for the chitinase was 5.2. The optimum pH, optimum temperature, pH stability, and thermal stability of chitinase were pH 7, 50°С, pH 6-8, and 60°С.
YQ308 chitinase showed higher hydrolyzing activity against ethylene glycol chitin and little activity against glycol chitosan. There was no enzymatic activity on p-NP-GlcNAc. According to this nomenclature system, YQ308 chitinase is not a β-N-acetyl-hexosaminidases because it does not hydrolyze p-NP-GlcNAc. It could be an endochitinase since it catalyzes release of N-acetylchitotetraose from colloidal chitin. The purified chitinase inhibited the hyphal extension of the fungi F. oxysporum, F. solani, and P. ultimum. We interesting in cloning the chitinase gene to a yeast transformation system for overexpression and application in biocontrol system. According to this idea, we cloned the chitinase gene.
N-acetylchitohexaose and chitohexaose showed the immune-enhancing ability. However, its productioncost was too high to put in use. The chitooligosaccharide mixture from YQ308 purified chitinase at 0.065 mg/ml showed the highest enhancing activation the proliferation of cord blood mononuclear cells.
中文摘要------------------------------------------------------------------------1
英文摘要---------------------------------------------------------------------------2
壹、前言---------------------------------------------------------------------------4
貳、文獻整理-----------------------------------------------------------------------7
一、幾丁質之起源------------------------------------------------------------7
二、幾丁質及幾丁聚醣之構造--------------------------------------------7
三、幾丁質之物理和化學性質-------------------------------------------11
四、幾丁質的酵素性分解-------------------------------------------------13
五、幾丁質及幾丁聚醣之應用-------------------------------------------16
六、幾丁質酶分類及應用--------------------------------------------------17
七、N-乙醯幾丁寡醣與幾丁寡醣-----------------------------------------27
八、生物性農藥於生物防治之應用--------------------------------------31
參、材料與方法------------------------------------------------------------------34
一、實驗材料------------------------------------------------------------------34
二、實驗試藥------------------------------------------------------------------34
三、實驗方法------------------------------------------------------------------34
(一) 真菌抑制劑及幾丁質酶生產菌株之篩選與鑑定----------------37
1. 篩菌----------------------------------------------------------------------37
2. 發酵液之製備----------------------------------------------------------37
3. 生物分析法-------------------------------------------------------------37
4. 菌種鑑定----------------------------------------------------------------38
(二) 真菌抑制劑生產條件之探討及應用-------------------------------38
1. 蝦蟹殼粉、幾丁質及幾丁聚醣純度之測定----------------------38
(1) 測定殘留碳酸鈣--------------------------------------------------38
(2) 測定殘留蛋白質--------------------------------------------------38
2. 最適發酵條件之探討-------------------------------------------------39
(1) 最適蝦蟹殼粉、幾丁質及幾丁聚醣的測定------------------39
(2) 最適碳源種類的測定--------------------------------------------39
(3) 最適碳源濃度的測定--------------------------------------------39
(4) 最適氮源種類的測定--------------------------------------------39
(5) 最適氮源濃度的測定--------------------------------------------40
(6) 最適金屬離子種類的測定--------------------------------------40
(7) 最適金屬離子濃度的測定--------------------------------------40
(8) 最適培養時間的測定--------------------------------------------40
(9) 最適培養溫度的測定--------------------------------------------40
(10) 最適培養體積的探討-------------------------------------------40
(11) 最適培養基pH值-----------------------------------------------41
3. 發酵液性質及生物活性探討----------------------------------------41
(1) 發酵液酵素活性測試--------------------------------------------41
(2) 菌體及發酵上清液抗菌測試-----------------------------------44
4. 菌體固定化-------------------------------------------------------------45
(1) 菌體前處理--------------------------------------------------------45
(2) 製備幾丁聚醣溶液-----------------------------------------------46
(3) 製備固定化菌體--------------------------------------------------46
(4) 固定化菌體抑制真菌活性曲線--------------------------------46
(三) 幾丁質酶分離純化、生化性質及抑制真菌探討--------------46
1. 酵素純化---------------------------------------------------------------46
(1) 硫酸銨前處理-----------------------------------------------------46
(2) 硫酸銨沈澱--------------------------------------------------------47
(3) 離子交換層析-----------------------------------------------------47
(4) 膠體過濾層析-----------------------------------------------------47
2. 酵素生化性質及抑制真菌分析------------------------------------48
(1) 幾丁質酶活性之測定--------------------------------------------48
(2) 蛋白質濃度測定--------------------------------------------------48
(3) 分子量測定--------------------------------------------------------48
(4) 酵素作用最適溫度的測定--------------------------------------49
(5) 酵素熱安定性的測定-------------------------------------------49
(6) 酵素作用最適pH的測定--------------------------------------49
(7) 酵素pH安定性的測定-----------------------------------------49
(8) 等電點的測定-色層焦集法------------------------------------50
(9) 酵素對基質特異性-----------------------------------------------50
(10) 酵素水解產物的分析-------------------------------------------50
(11) 菌體之生長曲線及幾丁質酶生產活性---------------------50
(12) 對病原真菌分生孢子及菌絲生物量之抑制測定---------50
(四) 酵素基因之探討------------------------------------------------------51
1. 染色體DNA的製備---------------------------------------------------51
2. 聚合酶連鎖反應------------------------------------------------------52
3. 洋菜膠體電泳分析PCR產物---------------------------------------53
4. 從PCR產物純化DNA 片段----------------------------------------53
5. 利用勝任細胞的轉形快速篩選目標基因-------------------------53
6. 核苷酸序列比對分析-------------------------------------------------54
(五) 酵素水解產物於臍帶血之應用-------------------------------------54
1. 分離臍帶血中之單核細胞-------------------------------------------54
2. 測定經Ficoll分離臍帶血中單核細胞及幹細胞含量----------54
3. 分離水解產物中之N-乙醯幾丁寡醣------------------------------55
4. 幾丁寡醣混合液、幾丁六醣與N-乙醯幾丁六醣
對單核細胞之影響----------------------------------------------------55
(六) 蛋白酶分離純化、生化性質及抑制真菌探討------------------57
1. 酵素純化---------------------------------------------------------------57
(1) 硫酸銨前處理-----------------------------------------------------57
(2) 硫酸銨沈澱--------------------------------------------------------57
(3) 離子交換層析-----------------------------------------------------57
(4) 膠體過濾層析-----------------------------------------------------57
2. 酵素生化性質及抑制真菌分析------------------------------------57
肆、結果與討論------------------------------------------------------------------58
(一) 真菌抑制劑及幾丁質酶生產菌株之篩選與鑑定----------------58
(二) 真菌抑制劑生產條件之探討及應用-------------------------------63
1. 蝦蟹殼粉、幾丁質及幾丁聚醣純度之測定---------------------63
2. 最適發酵條件之探討-------------------------------------------------64
(1) 最適蝦蟹殼粉、幾丁質及幾丁聚醣的測定------------------64
(2) 最適碳源種類的測定--------------------------------------------66
(3) 最適碳源濃度的測定--------------------------------------------66
(4) 最適氮源種類的測定--------------------------------------------66
(5) 最適氮源濃度的測定--------------------------------------------67
(6) 最適金屬離子種類的測定--------------------------------------67
(7) 最適金屬離子濃度的測定--------------------------------------67
(8) 最適培養時間的測定--------------------------------------------74
(9) 最適培養溫度的測定--------------------------------------------74
(10) 最適培養體積的探討-------------------------------------------74
(11) 最適培養基pH值-----------------------------------------------74
3. 發酵液性質及生物活性探討----------------------------------------79
(1) 發酵液酵素活性測試-------------------------------------------79
(2) 菌體及發酵上清液抗菌測試-----------------------------------79
4. 菌體固定化-------------------------------------------------------------84
(三) 幾丁質酶分離純化、生化性質探討及其應用-----------------90
1. 酵素純化---------------------------------------------------------------90
2. 酵素生化性質及抑制真菌分析------------------------------------90
(1) 分子量測定--------------------------------------------------------90
(2) 酵素作用最適溫度的測定--------------------------------------95
(3) 酵素熱安定性的測定-------------------------------------------95
(4) 酵素作用最適pH的測定--------------------------------------95
(5) 酵素pH安定性的測定-----------------------------------------95
(6) 等電點的測定-色層焦集法-----------------------------------100
(7) 酵素對基質特異性----------------------------------------------100
(8) 酵素水解產物的分析------------------------------------------100
(9) 菌體之生長曲線及幾丁質酶生產活性--------------------101
(10) 對病原真菌分生孢子及菌絲生物量之抑制測定--------101
(四) 酵素基因之探討-----------------------------------------------------106
(五) 酵素水解產物於臍帶血之應用------------------------------------111
1. 分離臍帶血中之單核細胞-----------------------------------------111
2. 測定經Ficoll分離臍帶血中單核細胞及幹細胞含量---------111
3. 幾丁寡醣混合液、幾丁六醣與N-乙醯幾丁六醣
對單核細胞之影響--------------------------------------------------111
(六) 蛋白酶分離純化、生化性質探討及其應用--------------------118
1. 酵素純化--------------------------------------------------------------118
2. 酵素生化性質及抑制真菌分析-----------------------------------118
(1) 分子量測定-------------------------------------------------------118
(2) 酵素作用最適溫度的測定-------------------------------------118
(3) 酵素作用最適pH的測定--------------------------------------125
伍、結論--------------------------------------------------------------------------128
王三郎,2000。生物技術。高立出版社。
高穗生,2000。微生物殺蟲劑之現況。2000年台北國際綠色環保研討會論文集。P. 4-30。
孫守恭,黃振文。1996。台灣植物鎌胞菌病害。世維出版社。
陳天,張皓冰,葉秀蓮。1991。殼聚醣常溫保鮮獼猴桃的研究。食品科學,10 : 34-36 .
蔡竹固。1995。植物病理學精要。台灣復文興業出版。
羅朝村,2000。生物性農業藥劑研發與應用。生物資源‧生物技術。2 (3): 9-10。
Aiba, S. 1993. Sudies on chitosan:Relationship between N-acetyl group distribution pattern and chitinase digestibility of partially N-acetylated chitosans. Int. J. Biol. Macromol. 15:241-245.
Allan, G. G. and Peyron., M. 1995. Molecular weight manipulation of chitosan. I:Kinetics of depolymerization by nitrous acid. Carbohydr. Res. 277:257-272.
Angel, Z.H., and Angel, A.B. 1983. chitinolytic activity from Neurosproa crassa. J. Gen. Microbiol. 129:3319-3323 .
Araki, Y.,and E. Ito. 1974 ,A pathway of chitosan formation in Mucor rouxii:enzymatic deacetylation of chitin, Biochem. Biophys. Res. Commun. 56:669-675.
Bagnasco, P., Fuente, L. D. L., Gualtieri, G., Noya, F. and Arias, A. 1998. Fluorescent Pseudomonas spp. as biocontrol agent against forage legume root pathogenic fungi. Soil Biol. Biochem. 30: 1317-1322.
Bassler, B.L., Yu, C., Lee, A.M., and Roseman, S. 1991. Chitin utilization by marine bacteria, degradation and catabolism of chitin oligosacchides by Vibrio furnissii . J. Biol. Chem. 266 (36):242-246.
Benhamou, N., Broglie, R. and Chet, I. 1993. Antifungal effect of bean endochitinase on Rhizoctonia solani:ultrastructal changes and cytochemical aspects of chitin breakdown. Can. J. Microbiol. 39:318-328.
Bhushan, B. 2000. Production and chacterization of a thermostable chitinase from a new alkalophilic Bacillus sp. BG-11. J. Appl. Microbiol. 88:800-808.
Bihari-Varga, M., Sepulchre, C., and Moczar , E. 1975 . Thermoanalytical studies on Protein -poly-saccharide complex of connective tissue. J. Thermal Anal. 7:675-678.
Blakeman, J. P. 1993. Pathogens in the foliar environment. Plant Pathol. 42:479-493.
Boller, T. and Mauch, F. 1988. Colorimetric assay for chitinase. In Methods in Enzymology. 161: 424-426, Academic Press, New York.
Bough, W.A. and Landes, D.R. 1977. Recovery and Nutritional evaluation of proteinaceous solid separation from whey by coagulation with chitosan . J. Dairy Sci. 59:1874-1877.
Bruijn, F. J. and O’Gara, F. 1997. Biological control of Pythium ultimum by Stenotrophomonas maltophilia W81 is mediated by an extracellular proteolytic activity. Microbiol. 143:3921-3931.
Budi, S. W., Tuinen, D., van, Arnould, C., Dumas-Gaudot, E., Gianinazzi-Pearson, V. and Gianinazzi, S. 2000. Hydrolttic enzyme activity of Paenibacillus sp. Strain B2 and effects of the antagonistic bacterium on cell integrity of two soil-brone pathogenic fungi. Appl. Soil Ecol. 15:191-199.
Cao, L-Z. and Lin, Z-B. 2002. Regulation on maturation and function of dedritic cells by Ganoderma lucidum polysaccharides. Immunol. Lett. 83:163-169.
Carroad , D.A., and Tom , R.A. 1978. Bioconversion of shellfish chitin waste:process conception and selection of microorganism . J. Food Sci. 43:1158-1161.
Chang, C. C. 1998. Cord blood stem cell transplantation-facts behind the media hype. Clinician Rev. 8:67-70.
Chemin L, Ismailov A, Haran S, and Chet I., 1995.Chitinolytic Enterobacter agglomerans antagonistic to fungal plant pathogens. Appl. Environ. Microbiol. 61:1720-6.
Chemin, L., Ismailov, Z., Haran, I. and Chet, I. 1995. Chitinolytic Enterobacter agglomerans antagonistic to fungal plant pathogens. Appl. Environ. Microbiol. 61:1720-1726.
Ciopraga, J. Gozia, O., Tudor, R., Brezuica, L. and Doyle, R. J. 1999. Fusarium sp. growth inhibition by wheat germ agglutinin. Biochim. et Biophys. Acta. 1428:424-432.
Civin, C. I. and Gore, S. D. 1993. Antigenic analysis of hematopoiesis:a review. J. Hematother. 2:137-144.
Cohen-Kupiec, R. and Chet, J. 1998. The molecular biology of chitin digest. Curr. Opin. Biotech. 9:270-277.
Collinge, D. B., Kragh, K. M., Mikkslesen, J. D., Nielsen, K. K., Rasmussen, U. and Vad, K. 1993. Plant chitinase. Plant J. 3:31-40.
Correa, J.U., Elango, I., and Polacheck, E. 1982. Endochitinase , a mannan - associated enzyme from Saccharomyces cerevisiae. J. Biol. Chem. 257:1392-1396.
Cosio, I. G., Fisher, R. A. and Carroad, D. A. 1982. Bioconversion of shellfish chitin waste: waste pretreatment, enzyme production, process design, and economic analysis. J. Food Sci. 47:901-906.
Deane, E. E., Whipps, J.M., Lynch, J. M. and Peberdy, J. F. 1998. The purification and characterization of a Trichoderma harzianum exochitinase. Biochem. Biophys. Acta. 1383:101-110.
Deshpande, M.V. 1986. Enzymatic degradation of chitin and its biological application. J. Sci. Ind. Res. 45:273-275.
Dickinson, K., Keer, V., Hitchcock, C.A. and Asams, D.J. 1989. Chitinase activity from Candida albicans its inhibition by allosamidin, J. Gen. Microbiol. 135:1417-1421.
Di Pietro, A., Lorito, M., Hayes, C. K., Brodway, R. M. and Harman, G. E. 1993. Endochitinase from Gliocladium virens:isolation, characterization, and synergistic antifungal activity in combination with gliotoxin. Phytopathol. 83:308-13.
Domard, A. and Cartier, N. 1989. Glucosamine oligomers:Preparation and characterization. Int. J. Biol. Macromol. 11:297-302.
Dunne, C., Crowley, J. J., Moenne-Loccoz, Y., Dowling, D. N., Bruijn, F. J. and O’Gara, F. 1997. Biological control of Pythium ultinum by Stenotrophomonas maltophilia W 18 is mediated by an extracellular proteolytic activity. Microbiol. 143:3921-3931.
Dunne, C., Moenne-Loccoz, Y., de Bruijin, F. J. and O’Gara, F. 2000. Overproduction of an inducible extracellular serine protease improves biological control of Pythium ultimum by Stenotrophomonas maltophilia strain W81. Microbiol. 146:2069-2078.
Dunne, C., Moenne-Loccoz, Y., McCarthy, J., Higgins, P., Powell, J., Dowling, D. N. and O’Gara, F. 1998. Combining proteolytic and phloroglucinol-producing bacteria for improved biocontrol of Pythium-mediated damping-off sugar beet. Plant Pathol. 47:299-307.
Elad, Y. 1994. Biological control of grape grey mould by Trichoderma harzianum. Crop Protect. 13:35-38.
Elad, Y. 1996. Mechanisms involved in the biological control of Botrytis cinerea incited diseases. Eur. J. Plant Pathol. 102:719-732.
Elad, Y. 1999. Role of Trichoderma harzianum protease in the biocontrol of Botrytis cinerea. Eur. J. Plant Pathol. 105:177-189.
Elad, Y. 2000. Biological control of foliar pathogens by means of Trichoderma harzianum and potential modes of action. Crop Protect. 19:704-714.
Fedi, S., Tola, E., Moenne-Loccoz, Y., Dowling, D. N., Smith, L. M. and O’Gara, F. 1997. Evidence for signalling between the phytopathogenic fungus Pythium ultimum and Pseudomonas fluorescens F113:P. ultimum represses the expression of genes in P. fluorescens F113, resulting in altered ecological fitness. Appl. Environ. Microbiol. 63:4261-4266.
Fenton, D.M., Eveleigh, D.E. 1981. Purification and mode of action of a chitosonase from Penicillium islandicum. J. Gen. Microbiol. 126:151-154 .
Flach, J., Pilet, P.-E. and Jolles, P. 1992. What’s new in chitinase reach ? Experientia. 48:701-716.
Flores-Guzmán, P., Gutiérrez-Rodríguez, M. and Mayani, H. 2002. In vitro proliferation, expansion, and differentiation of a CD34+ cell-enriched hematopoietic cell population from human umbilical cord blood in response to recombinated cytokines. Arch. Medical Res. 33:107-114.
Fuchs, E. and Segre, J. A. 2000. Stem cells:A new lease on life. Cell 100:143-155.
Gal, S., Choi, JY., Kim, CY., Cheong, YH., Choi, YJ., Lee, SY., Bahk, JD. And Cho, MJ. 1998. Cloning of the 52-KDa chitinase gene from Serratia marcescens KCTC2172 and its proteolytic cleavage into an active 35-KDa enzyme. FEMS microbiol. Lett. 160:151-158.
Gooday, G. W. 1990. Physiology of microbial degradation of chitin and chitosan. Biodegradation 1: 177-190.
Gorbach, V. I., Krasikova, I. N., Lukyanov, P. A., Loenko, Y. N., Soloveva, T. F., Ovodov, Y. S., Deev, V. V. and Pimenov, A. A. 1994. New glycolipids (chitooligosaccharide derivatives) possessing immunostimulating and antitumor activities. Carbohydr. Res. 260:73-82.
Harman, G. E., Hayes, C. K., Lorito, M., Broadway, R. M., DiPietro, A., Peterbaues, C. and Tronsmo, A. 1993. Chitinolytic enzymes of Trichoderma harzianum: purification of chitobiosidase and endochitinase. Phytopathol. 83:313-318.
Harish, S., Manjula, K., and Podile, A. R. 1998. Fusarium udum is resistant to themycolytic activity of biocontrol strain of Bacillus subtilis AF 1. FEMS Microbiol. Ecol. 25:385-390.
Hasegawa, M., Isogai, A. and Onabe, F. 1993. Preparation of low-molecular-weight chitosan using phosphoric acid. Carbohydr. Polym. 20:279-283.
Hashimoto, M., Ikegami, T., Seino, S. Ohuchi, N., Fukada, H., Sugiyama, J., Shirakawa M. and Watanabe, T. 2000. Expression and characterization of the chitin-binding domain of chitinase A1 from Bacillus circulans WL-12. J. Bacteriol. 182: 3045-3054.
Healy, L., May, G., Gale, K. and Greaves, M. F. 1995. The stem cell antigen CD34 functions as a regulator of hematopoietic cell adhesion. Proc. Natl. Acad. Sci. USA 92:12240-12244.
Helistö, P., Aktuganov, G., Galimzianova, N., Melentjev, A. and Korpela, T. 2001. Lytic enzyme complex of an antagonistic Bacillus sp. X-b: isolation and purification of components. J. Chromatogr. 758:197-205.
Hirano, S., Yamamote, T., Hayashi, M., Nishida, T. and luni, H. 1990. Chitinase activity in seeds coated with chitosan derivatives. Agric. Biol. Chem. 4:2719-2720.
Horikoshi, K. and Iika, S. 1959. Effect of lytic enzyme from Bacillus circulans and chitinase from Streptomyces sp. on Aspergillus oryzae. Nature 183:196-187.
Imoto,T., Yagishita ,K. 1971 . A simple activity measurement of lysozyme. Agric. Biol. Chem. 35:1154-1158 .
Inbar, J. and Chet, I. 1991. Evidence that chitinase produced by Aeromonas caviae is involved in the biological control of soil- borne plant pathogens by this bacterium. Soil. Biol. Biochem. 23:973-978.
Jeuniaux, C. 1966. Methods in enzymology. 8:644-654. Academic Press, New York.
Joshi, S., Kozlowski, M., Selvaraj, G., Iyer, V. N. and Davies, R. W. 1988. Cloning of the gene of the chitin utilization regulon of Serratia liquefacients. J. Bacteriol. 170:2984-2988.
Knorr, D. 1982. Functional properties of chitin and chitosan . J. Food Sci. 47:593-596 .
Knorr, D. 1983. Dye binding properties of chitin and chitosan. J. Food Sci. 48:36-39.
Knorr, D. 1984. Use of chitinous polymer in food. Food Technol. 1:85-88.
Knorr, D., Wampler, T. P., and Teutonico, R. A. 1985. Formation of pyrazines by chitin pyrolysis. J. Food Sci. 50:1762-1765.
Kobayashi, M., Watanabe, T., Suzuki, S. and Suzuki, M. 1990. Effect of N-acetylchitohexaose against Candida albicans infection of tumorbearing mice. Microbiol. Immunol. 34:413-426.
Kobayashin, D. Y., Guglielomoni, M. and Clarke, B. B. 1995. Isolation of the chitinolytic bacteria Xanthomonas maltophilia and Serratia marcescens as biological control agents for summer patches disease of turfgrass. Soil. Biol. Biochem. 27:1479-187.
Koga, D., Tsukamoto, T., Sueshige, N., Usumi, T. and Ide, A. 1989, Kinetics of chitinase from yam, Dioscorea oppsita thunb, Agric. Biol. Cnem. 53:3121-3126.
Koga, D., Mizuki, K., Ide, A., Kono, M.,Matsui, T. and Shimizu, C. 1990, Kinetics of a chitinase from a prawn, Penaeus japonicus. Agric. Biol. Chem. 54:2505-2512.
Koga, D., T. Hirata, N. Sueshige, S. Tanaka, and A. Ide.1992, Induction patterns of chitinases in yam callus by inoculation with autoclaved Fusarium oxysporium, ethylene, and chitin and chitosan oligosaccharides, Biosci. Biotech. Biochem. 56:280-285.
Lewis, I. D., Almeida-Porada, G., Du, J., Lemischka, I., Moore, K. A., Zanjani, E. D. and Verfaillie, C. M. 2001. Umbilical cord blood cells capable of engrafting in primary, secondary, and tertiary xenogeneic hosts are preserved after ex vivo culture in a noncontact system. Blood 97:3441-3449.
Lim, H. S., Kim, Y. S. and Kim, S. D. 1991. Pseudomonas stutzeri YPL-1 genetic transformation and antifungal mechanism against Fusarium solani, an agent of plant root rot. Appl. Environ. Microbiol. 57:510-516.
Lorito, M., Di Pietro, A., Hayes, C. K., Woo, S. L. and Harman, G. E. 1993a. Antifungal, synergistic interaction between chitinolytic enzymes from Trichoderma harzianum and Enterobacter cloacae. Phytopathol. 83:721-728.
Lorita, M., Harman, G. E., Hayes, C. K., Broadway, R. M., Tronsmo, A., Woo, S. L. and Di Pietro, A. 1993b. Chitinolytic enzymes produced by Trichoderma harzianum: antifungal activity of purified endochitinase and chitobiosidase. Phytopathol. 83:302-307.
Mabuchi, N. Hashizume, I. and Araki, Y. 2000. Characterization of chitinases excreted by Bacillus cereus CH. Can. J. Microbiol. 46:370-375.
Mahadevan, B. and Crawford, D. L. 1997. Properties of the chitinase of the antifungal biocontrol agent Streptomyces lydicus WYEC 108. Enzyme Microb. Technol. 20:489-493.
Marco, J. L. De. and Felix, C. R. 2002. Characterization of a protease produced by a Trichoderma harzianum isolate which controls cocoa plant witches'' broom disease. BMC Biochem. 3:1-7.
Mckay, R. 2000. Stem cells-hype hope. Nature 406:361-364.
McNiece, I. K. 2001. Ex vivo expansion of hematopoietic cells: what is the clinical need ? J. Hematother. Stem Cell Res. 10:431—433.
Miller, G. L. 1959. Use of dinitrosalicyclic acid reagent for determination of reducing sugar. Anal. Chem. 31:426-428.
Molano, J., Puran, A. and Cabib, E. 1977. A rapid and sensitive assay of chitinase using tritiated chitin. Anal. Biochem. 83: 648.
Moneral, J. and Reese, E. T. 1969. The chitinase of Serratia marcescens. Can. J. Microbiol. 15: 689-696.
Morimoto, K., Karita, S., Kimura, T., Sakka, K. and Ohmiya, K. 1999. Sequencing, expression, and transcription analysis of the Clostridium paraputrificum chiA gene encoding chitinase ChiA. Appl. microbiol. Biotechnol. 51:340-347.
Murao, S., Kawada, T., Itoh, H., Oyama, H. and Shin, T. 1992. Purification and characterization of a novel type of chitinase from Vibrio alginolyticus TK-22. Biosci. Biotech. Biochem. 56: 368-369.
Murata, T. a. U., T. 1997. Preparation of oligosaccharides units library and its utilization. Biosci. Biotech. Biochem. 61:1059-1066.
Nam, M. Y. Shon, Y. H. Kim, S. K. Kim, C. H. Jeong, T. R. and Nam, K. S. 2000. Effect of chitosan oligosaccharides on polyamine metabolism for chemopreventive activity. J. Chit. Chitos. 5:15-18.
Nam, M. Y., Shon, Y. H. Kim, S. K., Kim, C. H. and Nam, K. S. 1999. Inhibitory effect of chitosan oligosaccharides on the growth of tumor cells. J. Chit. Chitos. 4:184-188.
Ohtakara, A. 1961. Studies on the chitinolytic enzymes of black-kogi mold. Part I. Viscometric determination of chitinase activity by application of glycol chitin as a new substrate. Agric. Biol. Chem. 25(1): 50-54.
Ordentilich, A., Elad, Y. and Chet, I. 1988. The role of chitinase of Serratia marcescens in biocontrol of Sclerotium rolfsii. Phytopathol. 78:48-88.
Orkin, S. H. 2000. Stem cell alchemy. Nature Med. 6:1212-1213.
Overdijk, B. and Steijk, G. J. V. 1994. Human serum contains a chitinase: identification of an enzyme, formerly described as4-methylumbelliferyl-tetra-N-acetylchitotetraoside hydrolase (MU-TACT hydrolase). Glycobiol. 4:797-803.
Patil, R. S., Ghormade, V., and Deshpande, M. V. 2000. Chitinolytic enzymes: an exploration. Enzyme Microbial Technol. 26:473-483.
Pinelli, S. A., Toledo, G. A. R., Esquerra, B. I. R., Luviano, S. A. R. and Higuera, C. I. 1998. Methods for extracting chitin from shell waste. Archivos Latinoamericanos de Nutrition 48(1):58-61.
Pleban, S., Chernin, L. and Chet, I. 1997. Chitinolytic activity of an endophytic strain of Bacillus cereus. Lett. Appl. Microbiol. 25:284-288.
Podile, A. R. and Prakash, A. P. 1996. Lysisi and biological control of Aspergillus niger by Bacillus subtilis AF 1. Can. J. Microbiol. 42:533-538.
Podile, A. R. and Prakash, A. P. 1996. Lyses and biological control of Aspergillus niger by Bacillus subtilis AF 1. Can. J. Microbiol. 42:533-538.
Prenner, E. J., Lewis, R. N.A.H. and McElhaney, R. N. 1999. The interaction of the antimicrobial peptide gramicidin S with lipid bilayer model and biological membranes. Biochem. Biophysnol. Acta. 1462:201-221.
Roberts, G. A. F. 1992. Chitin Chemistry. MacMillan Press, London.
Roberts, W.K., and C.P. Selitrennidkoff. 1988. Plant and bacterial chitinases differ in antifungal activity. J. Biol. Chem. 134:169-176.
Roberts, W. K. and Selitrennikoff, C. P. 1988. Plant and Bacterial Chitinase Differ in Antifungal activity. J. General Microbiol. 134:169-176.
Roby, D., Gadell, A. and Toppan, A. 1987. Chitin oligosaccharides as elicitors of chitinase activity in melon plants. Biochem. Biophys. Res. Commun. 143:885-887.
Roffey, P. E. a. P., J. M. 1990. Cloning and expression of an Aeromonas hydrophila chitinase gene in Escherichia coli. Curr. Microbiol. 21:329-337.
Sakai, K., Yokota, A., Kurokawa, H., Wakayama, M. and Moriguchi, M. 1998. Purification and characterization of three thermostable endochitinases of a noble Bacillus strain, MH-1, isolated from chitin-containing compost. Appl. Environ. Microbiol. 64:3397-3402.
Sampson, M. N. and Gooday, G. W. 1996. A novel chitinase assay using the fluorescent brightener Calcofluor White M2R. In Chitin Enzymol. 2:227-234.
Shapira, R., Ordentlich, A., Chet, I. and Openheim, A. B. 1989. Control of plant diseases by chitinase expressed from cloned DNA in Escherichia coli. Phytopathol. 79:1246-1249.
Shigemasa, Y., Saito, K., Sashiwa, H. and Saimoto, H. 1994. Enzymatic degradation of chitins and partially deacetylated chitins. Int. J. Biol. Microbiol. 16:43-49.
Singh, P. P., Shin, Y. C., Park, C. S. and Chung, Y. R. 1999. Biological control of Fusarium wilt of cucuber by chitinolytic bactera. Phytopathol. 89:92-99.
Sneh, B. 1981. Use of rhizosphere chitinolytic bacteria for biological control of Fusarium oxysporum f. sp. diantibi in carnation. Phytopath. 100:251-256.
Stanley, W.L., Watters, G.G., Chan, B.G., Kelly,S. H. 1976. Immobilization of glucose isomerase on chitin with glutaraldehyde and by simple adsorption. Biotechnol. Bioeng. 18:439442 .
Stanly, W. L., Watter, G. G. Chan, B. G. and Marcer. J. M. 1975. Lactose and other enzymes bound to chitin with glutaldehyde. Biotechnol. Bioeng. 17:315-318.
Streichsbier, F. 1983. Utilization of chitin as sole carbon and nitrogen source by Chrommbacterium violaceum. FEMS Microbiol. Lett. 19:129-132.
Suzuki, K., Mikami, T., Okawa, Y., Tokoro, A., Suzuki, S. and Suzuki, K. 1986. Antitumor effect of hexa-N-acetylchitohexaose and chitohexaose. Carbohy. Res. 151:403-408.
Tagawa, K. and Okazaiki, K. 1991. Isolation and some culture conditions of Streptomyces species which produce enzyme lysing Aspergillus niger cell wall. J. Ferment. Bioeng. 71:230-236.
Takayanagi, T., Ajisaka, K., Takiguchi, Y. and Shimahara, K. 1991. Isolation and characterization of thermostable chitinases from Bacillus licheniformis X-7u. Biochem. Biophys. Acta. 1078:404-410.
Takegawa, K., Mikami, B., Iwahara, S., Marita, Y., Yamamoto, K. and Tochikura, T. 1991. Complete amino acid sequence of endo-β-N-acetylglucosaminidase from Flavobacterium sp. Eur. J. Biochem. 202:175-180.
Timmis, K., Hobbs, G. and Berkeley, R. C. W. 1974. Chitinolytic Clostridia isolated from marine mud. Can. J. Microbiol. 20:1284-1285.
Todd, E. W. 1949 Quantitative studies on the total plasmid and trypsin inhibitor of human blood serum. J. Exptl. Med. 39:295-308.
Tokro, A., Suzuki, K. and Matsumoto, T. 1998. Chemotactic response of human neutrophils to N-acetylchitohexaose in vitro. Microbiol. Immunol. 32:387-394.
Trachuk, L. A., Shemiakina, T. M., Chestukhina, G. G. and Stepanov, V. M. 1996. Bacillus cereus chitinases:isolation and characteristics. Biokhimiia. 61:357-368.
Tsujibo, H., Minoura, K., Miyamoto, K., Endo, H., Moriwaki, M. and Inamori, Y. 1993. Purification and properties of a thermostable chitinase from Streptomyces thermoviolaceus OPC-520. Gene. 134: 113-117.
Tsutomu, T. Kasumi, A., Yasuyaki, T. and Venzo, S. 1991. Isolation and characterization of thermostable chitinase from Bacillus licheniformis. Biochem. Biophy. Acta. 1078:404-410.
Usui, T., Hayashi, Y., Nanjo, F., Sakai, K. and Ishido, Y. 1987. Transglycoylation reaction of a chitinase purified from Nocardio orientalis. Biochem. Biophys. Acta. 9923:302-309.
Wang, S. L. and Chang, W. T. 1997a. Purification and characterization of two bifunctional chitinase/lysozymes extracellularly produced by Pseudomonas aeruginosa K-187 in a shrimp and crab shell powder medium. Appl. Environ. Microbiol. 63:380-386.
Wang, S. L., Chio S. H. and Chang, W. T. 1997b. Production of chitinase from shellfish waste by Pseudomonas aeruginosa K-187. Proc. Natl. Sci. Counc. ROC (B), 21:71-78.
Wang, S.Y., Moyne, A., Thottappilly, G., Wu, S. Locy, R. D. and Singh, N. K. 2001. Purification and characterization of a Bacillus cereus exochitinase. Enzyme Microb. Technol. 28:492-498.
Watanabe, T., Oyanagi, W., Suzuki, K. and Tanaka, H. 1990. Chitinase system of Bacillus circulans WL-12 and importantance of chitinase A1 in chitin degradation. J. Bacteriol. 172:4017-4022.
Watanabe, T., Kobori, K., Miyashita, K., Fujii, T., Sakai, H., Uchida, M. and acid 2000 in chitinase A1 of Bacillus circulans WL-12 as essential residues for chitinase activity. J. Biol. Chem. 268:18567-18572.
Wortman, A. T., Somerville, C. C. and Colwell, R. R. 1986. Chitinase determinants of Vibrio vulnificus:gene cloning and applications of a chitinase probe. Appl. Environ. Microbiol. 52:142-145.
Wynne, E. C. and Pemberton, J. M. 1986. Cloning of a gene cluster from Cellvibrio mixtures which codes for cellulase, chitinase, amylase and pectinase. Appl. Environ. Microbiol. 52:1362-1367.
Yabuki, M., Mizushina, K., Amatatsu, T., Ando, A., Fujii, T., Shimada, M. and Yamashita, M. 1986. Purification and characterization of chitinase and chitobiase produced by Aeromonas hydropila subsp.anaerogenes A52. J. Gen. Appl. Microbiol. 32:25-38.
Yang, T. and Zall, R.R. 1984. Chitosan membranes for reverse osmosis application. J. Food Sci. 49:91-96.
Yu, C., Lee, A.M., Bassler, B.L., and Roseman, S. 1991. Chitin utilization by marine bacteria, a physiological function for bacterial adhesion to immobilized carbohydrate. J. Biochem. 266 : 260-266 .
Zhu, B. C., Lo, J. Y., Li, Y. T., Li S. C., Jaynes, J. M., Gildemeister, O. S., Laine, R. A. and Ou, C. Y. 1992. Thermostable, salt tolerant, wide pH range novel chitobioase from Vibrio parahaemolyticus : isolation, characterization, molecular cloning, and expression. J. Biochem. 112 : 163-167.
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