Title

以Aspergillus niger NCH-318生產聚甘露糖酶最適條件及誘導特性探討暨聚甘露糖水解產物之益生效果

Translated Titles

Optimization of mannanase production and induction properties from Aspergillus niger NCH-318 and the prebiotic effect of mannan hydrolysis products

Authors

黃麗螢

Key Words

聚甘露寡糖 ; 益生質 ; 益生菌 ; 聚甘露糖酶 ; probiotics ; mannooligosaccharide ; mannanase ; prebiotics

PublicationName

中臺科技大學食品科技研究所學位論文

Volume or Term/Year and Month of Publication

2010年

Academic Degree Category

碩士

Advisor

林澤群

Content Language

繁體中文

Chinese Abstract

本研究探討使用具有聚甘露糖酶活性之Aspergillus niger NCH-318為試驗菌株,以去油椰子粕為碳源並以反應曲面法( response surface method, RSM)探討聚甘露糖酶(mannanase)生產培養基組成的最適化。此外,以不同醣類做為培養Asp. niger NCH-318探討酵素誘導特性,最後以部分純化的聚甘露糖酶製備聚甘露糖之水解產物,再以水解產物進行乳酸菌生長試驗。 Asp. niger NCH-318經RSM試驗,得到生產聚甘露糖酶之最適培養條件結果如下:去油椰子粕濃度3.12% (w/v)、酵母萃取物8.67(g/L)、起始pH 4.6,培養溫度40℃,接種量為106 spores/mL,培養於第六天有最佳的聚甘露糖酶活性(約236.64 U/mL)。 利用酵素水解聚甘露糖所得產物誘導Aspergillus niger NCH-318生產聚甘露糖酶效果顯著。多醣則以蒟蒻粉、刺槐豆膠及關華豆膠誘導生產聚甘露糖酶之效果較顯著。 B. infantis BCRC 14602及B. angulatum BCRC 14665在甘露糖、聚甘露糖水解產物及刺槐豆膠培養期間,以聚甘露糖水解產物為碳源培養B. infantis BCRC 14602及B. angulatum BCRC 14665較其他兩碳源快進入對數生長期。利用可滴定酸估計培養基中乳酸含量,以聚甘露糖水解產物做為碳源培養B. infantis BCRC 14602及B. angulatum BCRC 14665之乳酸含量較高。

English Abstract

A strain Aspergillus niger NCH-318 capable of producing mannanase was investigated in this study. We used coconut meal as carbon source and other factors to optimize medium composition by response surface method. In addition, different sugars were useed as carbon sources to study mannanase induction for this strain. Finally, mannan hydrolysates were used to cultivate bifidobacteria to understand their growth-enhanceing effect. The optimum culture conditions for mannanase production were as follow: defatted coconut meal 3.12% (w/v), yeast extract 8.67 g/L, initial pH 4.6, incubation temperature 40℃, and inoculum size of 106 spores/mL. The best mannanase activity (236.64 U/mL) was obtained under the optimal condition after six days of cultivation. Mannooligosaccharide could induce mannanase production by this strain. The induction effect was significant. When we used the media containing mannooligosaccharide to culture B. infantis BCRC 14602, B. longum BCRC 14664 and B. angulatum BCRC 14665, the results showed B. infantis BCRC 14602 and B. angulatum BCRC 14665 grew faster than cultured by other carbon sources. The data showed that both B. infantis BCRC 14602 and B. angulatum BCRC 14665 spent less time to reach log phase of growth curve when they were cultured by mannooligosaccharide. The lactic acid production was also higher than those cultures without mannooligosaccharide. Therefor, the mannooligosaccharides could promote some bifidobacteria growth.

Topic Category 健康科學院 > 食品科技研究所
工程學 > 化學工業
Reference
  1. Ademark, P., Larsson, M., Tjerneld, F., and Stalbrand, H. 2001. Multiple α-galactosidases from Aspergillus niger: purification, characterizationand substrate specificities. Enzyme Microb. Technol. 29: 441-448.
    連結:
  2. Aspinall, G.O. 1959. Structural chemistry of the hemicelluloses. AdvCarbohydr. Chem. 14: 429-468.
    連結:
  3. Baurhoo, B., Phillip, L., and Ruiz-Feria, C. A. 2007. Effects of purified lignin and mannan oligosaccharides on intestinal integrity and microbial populations in the ceca and litter of broiler chickens. Poult. Sci. 86: 1070-1078.
    連結:
  4. Bruzzese, E., Volpicelli, M., Squaglia, M., Tartaglione, A., and Guarino, A. 2006. Impact of prebiotics on human health. Digestive and Liver Disease, 38(Suppl 2): S283-S287.
    連結:
  5. Bulpin, P. V., Gidley, M. J., Jeffcoat, R., and Underwood, D. J. 1990. Development of a biotechnological process for the modification of galactomannan polymers with plant α-galactosidase. Carbohydr. Polym. 12: 155-168.
    連結:
  6. Chandrasekaran, R., Radha, A., and Okuyama, K. 1998. Morphology of galactomannans: an X-ray structure analysis of guaran. Carbohydr. Res. 306: 243-255.
    連結:
  7. Chang, Y. T., Chen, Y. S., Lin, K. T., Lin, I. F., Chen, Y. F., Yen, Y. H., Liu, B. L., and Chang, Y. N. 2006. Characteristics of galactomannanase for degrading konjac gel. J. Mol. Catal. B: Enzym. 43: 153-157.
    連結:
  8. Dea, I. C. M., and Morrison, A. 1975. Chemistry and interactions of seed galactomannans. Adv. Carbohydr. Chem. Biochem. 31:241-312.
    連結:
  9. Dekker, R. F. 1985. Biodegradation of the hemicelluloses. In: Higuchi (ed) Biosynthesis and Biodegradation of Wood Component (pp 505-532). Academic Press, New York.
    連結:
  10. Dey, P. M. 1978. Biochemistry of plant galactomannans. Adv. Carbohydr. Chem. Biochem. 35: 341-376.
    連結:
  11. Dhawan, S. and Kaur, J. 2007. Microbial mannanases: an overview of production and applications. Crit. Rev. Biotechnol. 27(4): 197-216.
    連結:
  12. Do, B. C., Dang, T. T., Berrin, J. G., Haltrich, D., To, K. A., Sigoillot, J. C. and Yamabhai, M. 2009. Cloning, expression in Pichia pastoris, and characterization of a thermostable GH5 mannan endo-1, 4-β-mannosidase from Aspergillus niger BK01, Microb. Cell. Fact. 8: 59-70.
    連結:
  13. Duffaud GD, McCutchen CM, Leduc P, Parker KL, Kelly RM 1997. Purification and characterization of extremely thermostable β-mannanase, β-mannosidase, and α-galactosidase from the hyperthermophilic Eubacterium Thermotoga neapolitana 5068. Appl. Environ. Microbiol. 63: 169-177.
    連結:
  14. Duruksu, G., Ozturk, B., Biely, P., Bakir, U., and B. Ogel, Z. 2009. Cloning, expression and characterization of endo-β-1, 4-mannanase from Aspergillus fumigatus in Aspergillus sojae and Pichia pastoris. Biotechnol. Prog. 25(1): 271-276.
    連結:
  15. Gallaher, D. D., and J. Khil. 1999. The effect of synbiotics on colon carcinogenesis in rats. J. Nutr. 129(7 Suppl):1483S–1487S.
    連結:
  16. Gibson, G. R., and Roberforid, M. B. 1995. Dietary modulation of the human colonic microbiota: introducing the concept of prebiotics. J. Nutr. 125: 1401-1412.
    連結:
  17. Giovanni, M. 1983. Response surface methodology and product optimization. Food Technol. 37(11): 41-45, 83.
    連結:
  18. Heck, K. L., Hays, G., and Orth, R. J. 2003. Critical evaluation of the nursery role hypothesis for seagrass meadows. Mar. Ecol. Prog. Ser. 253: 123-136.
    連結:
  19. Heinrichs, M., Baumgartner, T., Kirschbaum, C., and Ehlert, U. 2003. Social support and oxytocin interact to suppress cortisol and subjective responses to psychosocial stress. Biol Psychiatry 54: 1389-1398.
    連結:
  20. Hofacre, C. L., Beacorn, T. S., Collett, S. and Mathis, G. 2003. Using competitive exclusion mannanoligosaccharide and other intestinal products to control necrotic enteritis. J. Appl. Poult. Res. 12: 60-64.
    連結:
  21. Holzapfel, W. H., Haberer, P., Snel, J., Schillinger, U., and Huis in’t Veld, J. H. J. 1998. Overview of gut flora and probiotics. Int. J. Food Microbiol. 41: 85-101.
    連結:
  22. Hongshu, Z., Jinggan, Y., and Yan, Z. 2002. The glucomannan from ramie.Carbohydr. Polym. 47: 83-86.
    連結:
  23. Hossain, M. Z., Abe, J-I., and Hizukuri, S. 1996. Multiple forms of β-mannanase from Bacillus sp. KK01. Enzyme Microb. Technol. 18: 95-98.
    連結:
  24. Jeffries, T. W. 1994. Biodegradation of lignin and hemicelluloses, In: Ratledge C (ed.) Biochemistry of microbial degradation. Kluwer, Dordrecht, pp 233-277.
    連結:
  25. Jiang, Z. Q., Wei, Y., Li, D. Y., Li, L., Chai, P. P., and Kusakabe, I. 2006. High-level production, purification and characterization of a thermostable b-mannanase from the newly isolated Bacillus subtilis WY34. Carbohybr. Polym. 66: 88-96.
    連結:
  26. Kirk, O., Borchet, T. V., and Fuglsang, C. C. 2002. Industrial enzyme applications. Curr. Opin. Biotechnol. 13: 345-351.
    連結:
  27. Kobayashi, H., and Suzuki, H. 1972. Studies on the decomposition of raffinose by α-galactosidase of mold. J. Ferment. Technol. 50: 625-632.
    連結:
  28. Krishman, S., Prapulla S. G., Rajalakshmi, D., Misra, M. C. and Karanth, N. G. 1998. Screening and selection of media components for lactic acid production using Plackett-Burman design. Bioprocess Eng. 19: 61-65.
    連結:
  29. Krishna, C. 1999. Production of becterial cellulases by solids state bioprocessing of banana wastes. Biores. Technol. 69: 231-239.
    連結:
  30. Lee, Y. K., and Salminen, S. 1995. The coming of age of probiotics. Trends Food Sci. Technol. 6: 241-245.
    連結:
  31. Liepman, A. H., Nairn, C. J., Willats, W. G. T., Sorensen, I., Roberts, A. W., and Keegstra, K. 2007. Functional genomic analysis supports conservation of function among cellulose synthase-like A gene family members and suggest diverse roles of mannans in plants. Plant Physiol. 143: 1881-1893.
    連結:
  32. Lin, S. S., Dou, W. F., Xu, H. Y., Li, S. S., Xu, Z. H., and Ma, Y. H. 2007. Optimization of medium composition for the production of alkaline β-mannanase by alkaline Bcillus sp. N16-5 using response surface methodology. Appl. Microbiol. Biotechnol. 75: 1015-1022.
    連結:
  33. Lindberg, B., Lonngren, J., Ruden, U., and Simmons, D. A. R. 1973. A reinvestigation of Shigella Jlexneri O-antigens by methylation analysis. Eur. J. Biochem. 32: 15-18.
    連結:
  34. Ma, Y., Xue, Y., Dou, Y., Xu, Z., Tao, W., and Zhou, P. 2004. Characterization and gene cloning of a novel β-mannanase from alkailphilic Bacillus sp. N16-5. Extremophiles, 8: 447-454.
    連結:
  35. Malherbe, S., and Cloete, T.E. 2002 Lignocellulose biodegradation: fundamentals and applications: A review. Environ. Sci. Biotechnol. 1: 105-114.
    連結:
  36. Meier, H. (1958) On the structure of cell walls and cell wall mannans from ivory nuts and from dates. Biochim. Biophys. Acta 28: 229-240.
    連結:
  37. Meier, H., and Reid, J. S. G. (1982) Reserve Polysaccharides other than starch in higher plants. In: Loewus FA, Tanner W (eds) Encyclopedia of Plant. vol. 13A. Springer, Berlin, pp 418-471.
    連結:
  38. Millane, R. P., and Hendrixson, T. L. (1994) Crystal-structures of mannans and glucomannans. Carbohydr. Polym. 25: 245-251.
    連結:
  39. Moreira, L. R. S. and Filho, E. X. F. 2008. An overview of mannan structure and mannan-degrading enzyme systems. Appl. Microbilo. Biotechnol. 79: 165-178.
    連結:
  40. Myers, R. H. and Montagomery, D. C. 1995. Response surface methodology: process and product optimization using designed experiments. John wiley & Sons. Inc., New York.
    連結:
  41. Northcote, D. H. 1972. Chemistry of the plant cell wall. Annu. Rev. Plant. Physiol. 23: 113-132.
    連結:
  42. Ouwehand, A.C., Derrien, M., De Vos, WM., Tiihonen, K., and Rutonen, N. 2005. Prebiotics and other microbial substrates for gut functionality. Curr. Opin. Chem. Biol. 16: 212-217.
    連結:
  43. Parvathy, K. S., Susheelamma, N. S., Tharanathan, R. N. and Gaonkar, A.K. 2005. A simple non-aqueous method for carboxmethylation of galactomannans. Carbohydr. Polym. 62: 137-141.
    連結:
  44. Parvathy, K. S., Susheelamma, N. S., Tharanathan, R. N., and Gaonkar, A. K. 2005. A simple non-aqueous method for carboxmethylation of galactomannans. Carbohydr. Polym. 62: 137-141.
    連結:
  45. Perez, L., Mas, V., Coll, J., and Estepa, A. 2002. Enhanced detection of viral hemorrhagic septicemia virus (a salmonid rhabdovirus) by pretreatment of the virus with a combinatorial library-selected peptide. J. Virol. Methods 106: 17-23.
    連結:
  46. Perrin, S., Fougnies, C., Grill, J.P., Jacobs, H., and Schneider, F. 2002. Fermentation of chicory fructo-oligosaccharides in mixtures of different degrees of polymerization by three strsins of bifidobacteria. Can. J. Microbiol. 48: 759-763.
    連結:
  47. Petkowicz, C. L. O., Reicher, F., Chanzy, H., Taravel, F. R., and Vuong, R. 2001. Linear mannan in the endosperm of Schizolobium amazonicum. Carbohydr. Polym. 44: 107-112.
    連結:
  48. Petkowicz, C. L. O., Schaefer, S., and Reicher, F. 2007. The mannan from Schizolobium parahybae endosperm is not a reserve polysaccha-ride. Carbohydr. Polym. 69: 659-664.
    連結:
  49. Popa, V. I., and Spiridon, J. 1998. Hemicelluloses: structure and properties.In: Dumitriu S (ed) Polysaccharides: structural diversity and functional versatility. Marcel Dekker, New York, pp 297-311.
    連結:
  50. Preston, R. D. 1968. Plants without cellulose. Sci. Am. 216: 102-108.
    連結:
  51. Puls, J. and Schuseil, J. 1993. Chemistry of hemicelluloses: relationship between hemicelluloses structure and enzyme required for hydrolysis. In: Coughlan, M. P. and Hazlewood, G. P. (eds) Hemicellulose and hemicellulases. Portland, London, pp 1-27.
    連結:
  52. Sako, T., Matsumoto, K., and Tanaka, R. 1999. Recent progress on research and applications of non-digestible galactooligosaccharides. Int. Dairy J. 9: 69-80.
    連結:
  53. Samra, Z. Q. and Athar, M. A. 2008. Cloning, sequence, expression and characterization of human β-mannosidase. Acta Biochem. Pol. 55(3): 479-490.
    連結:
  54. Shobha, M. S., Kumar, A. B. V., Tharanathan, R. N., Koka, R., and Gaonkar, A. K. 2005. Modification of guar galactomannan with the aid of Aspergillus niger pectinase. Carbohydr. Polym. 62: 267-273.
    連結:
  55. Sittikijyothin, W., Torres, D., and Goncalves, M. P. 2005. Modelling the rheological behaviour of galactomannan aqueous solutions. Carbohydr. Polym. 59: 339-350.
    連結:
  56. Songsiriritthigul, C., Buranabanyat, B., Haltrich, D. and Yamabhai, M. 2010. Efficient recombinant expression and secretion of a thermostable GH26 mannan endo-1, 4-β-mannosidase from Bcillus licheniformis in Escherichia coli. Microb. Cell Fact. 9:20.
    連結:
  57. Staykov, Y., Spring, P., Denev, S., and Sweetman, J. 2007. Effect of a mannan oligosaccharide on the growth performance and immune status of rainbow trout (Oncorhynchus mykiss). Aquac. Int. 15: 153-161.
    連結:
  58. Talbot, G., and Sygusch, J. 1990. Purification and characterization of thermostable β-mannanase and α-galactosidase from Bacillus sterrothermophilus. Appl. Environ. Microbiol. 56(11): 3505-3510.
    連結:
  59. Tang, Z. R, Yin, Y. L., Nyachoti, C. M., Huang, R. L., Li T. J., Yang, C. B., Yang, X. J., Peng, J., Qi D. S., Xing,J. J., Sun Z. H., and Fan, M. Z. 2005. Effect of dietary supplementation of chitosan and galacto-mannan-oligosaccharide on serum parameters and the insulin-like growth factor-I mRNA expression in early-weaned piglets. Domest. Anim. Endocrinol. 28: 430-441.
    連結:
  60. Techapun, C., Poosaran, N., Watanabe, M., and Sasaki, K. 2003. Thermostable and alkaline-tolerant microbial cellulase-free xylanases produced from agricultural wastes and the properties required for use in pulp bleaching bioprocesses: a review. Process Biochem. 38: 1327-1340.
    連結:
  61. Timell, T. E. 1967. Recent progress in the chemsity of wood hemicelluloses. Wood Sci. Technol. 1: 45-70.
    連結:
  62. Tomasik, P.J. and Tomasil P. 2003. Probiotics and prebiotics. Cereal Chemistry, 80(2): 113-117.
    連結:
  63. Trafalska, E. and Grzybowski, A. 2006. Probiotics and prebiotics in prevention of chronic civilization diseases. New Medicine, 9:3-6.
    連結:
  64. Volek, Z., Marounek, M., and Skrivanova, V. 2007. Effect of a starter diet supplementation with mannan-oligosaccharide or inulin on health status, caecal metabolism, digestibility of nutrients and growth of early weaned rabbits. Animal, 1: 523-530.
    連結:
  65. Willfor, S., Sjoholm, R., Laine, C., Roslund, M., Hemming, J., and Holmbom, B. 2003. Characterisation of water-soluble galactoglucomannan from Norway spruce wood and thermomechanical pulp. Carbohydr. Polym. 52: 175-187.
    連結:
  66. Yoon, K. H., Chung, S., and Lim B. L. 2008.characterization of the Bacillus subtilis WL-3 mannanase from a recombinant Escherichia coli. J. Microbiol. 46(3): 344-349.
    連結:
  67. Zahura, U. A., Rahman, M. M., Inoue, A., Tanaka, H., and Ojima, T. 2010. An endo-β-1, 4-mannanase, AkMan, from the common sea hare Aplysia kurodai. Comp. Biochem. Physiol. B. 157:137-143.
    連結:
  68. 邱秉毅,2008,聚甘露糖酶生產菌之篩選及培養條件探討與酵素特性分析,中臺科技大學食品科技研究所碩士論文,台中。
    連結:
  69. Asenjo, J. A., and Patrick, I. 1990. Large-scale protein purification. In: HarrisELV, Angal S (eds) Protein purification applications: a practicalapproach. IRL, Oxford, pp 1-28.
  70. Coughlan, M. P., Tuohy, M. G., Filho, E. X. F., Puls, J., Claeyssens, M., Vrsanska, M., and Hughes, M. 1993. Enzymological aspects of microbial hemicellulases with emphasis on fungal systems. In: Coughlan MP (ed) Hemicellulose and hemicellulases. Portland, London, pp 53-84.
  71. Cristofaro, E., Mottu, F., and Wuhrmann, J. J. 1974. Involvement of the raffinose family of oligosaccharides in flatulence. In: Sipple HL, MCNutt K (eds) Sugars in nutrition. Academic, New York, pp 313-336.
  72. Filho, E. X. F. 1998. Hemicellulases and biotechnology. In: Pandalai SG(ed) Recent research developments in microbiology. Research Signpost, Trivandrum, pp 165-176.
  73. Kirk, K., and Cullen, D. 1998. Enzymology and molecular genetics of wood degradation by white rot fungi. In: Young RA, Akhtar M (eds) Environmental friendly technologies for pulp and paper industry. Wiley, New York, pp 273-307.
  74. McCleary, B. V. and Matheson, N. K. 1986. Polysaccharides having a β-D-xylan backbone. Adv. Carbohydr. Chem. Biochem. 44: 158-164.
  75. Sa-Pereira, P., Paveia, H., Costa-Ferreira, M., and Aires-Barros, M.R. 2003. A new look at xylanases. Mol. Biotechnol. 24: 257-281.
  76. Singh, S. K., Clarke, I. D., Terasaki, M., Bonn, V. E., Hawkins, C., Squire, J. and Dirks, P. B. 2003. Identification of a cancer stem cell in human brain tumors. Cancer. Res. 63: 5821-5828.
  77. Wong, K. K. Y., and Saddler, J. N. 1993 Applications of hemicellulases in the food, feed, and pulp and paper industries. In: Coughlan, M. P., and Hazlewood, G. P. (eds) Hemicellulose and hemicellulases. Portland, London, pp 127-143.
  78. 王三郎,1999,木質纖維素廢棄物之回收再利用,生物資源生物技術1(3)129:136。
  79. 王姿惠,2001,益生菌於含果寡糖之培養環境中生長情形之探討,國立中興大學食品科學系碩士論文,台中。
  80. 李晉嘉,2003,以反應曲面研究生化柴油之最優化酵素合成,大葉大學食品工程系,彰化。
  81. 林宛柔,2005。反應曲面法於綠藻Chlorella pyrenoidosa NCHU-6異營培養基組之最適化研究,國立中興大學食品科學系碩士論文,台中。
  82. 林澤揚,1996,Trichoderma longibrachiatum 185聚木糖酶之特性探討及半纖維素碳源對酵素誘導之影響,國立中興大學食品科學研究所碩士論文,台中。
  83. 林澤群,2003,Aspergillus niger NCH-189聚甘露糖酶之生產、純化、特性及其在低聚甘露糖製備之應用,國立中興大學食品科學系博士論文,台中。
  84. 洪哲穎、陳國誠,1992,回應曲面實驗設計法在微生物酵素生產上之應用,化工 39(2):3-18。
  85. 高馥君,1992,反應曲面在食品開發上的應用,食品工業 24(3):32-41。
  86. 陳清泉,1993,最適化實驗設計在食品工業產品開發上的應用。食品工業 25(2):50-62。
  87. 黎正中譯。Montgomery, D. C.著。1998。實驗設計與分析 第十四章:反應曲面法和其他製程最佳化途徑。高利圖書公司出版。
  88. 蘇祖良,2000,酵母菌 Saccharomyces diastaticus LORRE-316乙醇醱酵之最適化與狀態估計,中正大學化學工程研究所,嘉義。