Hostname: page-component-586b7cd67f-g8jcs Total loading time: 0 Render date: 2024-11-26T18:51:10.810Z Has data issue: false hasContentIssue false
  • English
  • Français

cDNA cloning and prokaryotic expression of β-glucosidase in tea plant [Camellia sinensis (L.) O. Kutze]

Published online by Cambridge University Press:  13 June 2008

Jiang Chang-Jun ,
Li Yuan-Hua  and
Fang Wan-Ping
Jiang Chang-Jun*
Affiliation:
Key Laboratory of Tea Biochemistry and Biotechnology, Ministry of Agriculture, Anhui Agricultural University, Hefei 230036, China
Li Yuan-Hua
Affiliation:
Key Laboratory of Tea Biochemistry and Biotechnology, Ministry of Agriculture, Anhui Agricultural University, Hefei 230036, China
Fang Wan-Ping
Affiliation:
Key Laboratory of Tea Biochemistry and Biotechnology, Ministry of Agriculture, Anhui Agricultural University, Hefei 230036, China
*
*Corresponding author. Email: [email protected]
Get access Rights & Permissions [Opens in a new window]

Abstract

The β-glucosidase gene has important effects on alcoholic aroma precursors and insect resistance of the tea plant [Camellia sinensis (L.) O. Kutze]. The complete cDNA sequence of β-glucosidase of the tea plant was cloned; its full length was 1475 bp, and shared 40–60% similarity with corresponding parts of the nucleotide sequence of β-glucosidase gene from other plants. Its secondary structure contains 14.33% α-helix, 25.43% β-pleated sheet and many functional amino acid domains. The β-glucosidase gene was cloned into the pET-32a expression system and expressed at high-efficiently in Escherichia coli BL21 (DE3); the molecular weight of expressed fusion protein was 63 kDa. The results of enzymic reaction showed that the fusion protein possessed normal bioactivity, and it could catalyse the dehydration of the glycosidic bond. The soluble fusion protein was expressed mainly in the cytoplasm.

Keywords

tea (Camellia sinensis)β-glucosidase; cDNAprokaryotic expression
Type
Research Article
Information
Chinese Journal of Agricultural Biotechnology , Volume 2 , Issue 2 , August 2005 , pp. 107 - 111
Copyright
Copyright © China Agricultural University and Cambridge University Press 2005

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Cheng, XD and Fujio, Y (1997) Purification and some properties of extracellular β-glucosidase from Rhizopus japonicus IFO5318. Acta Microbiological Sinica 37: 368–373 (in Chinese with English abstract).Google Scholar
Cicek, M (1998) Structure and expression of a dhurrinase (beta-glucosidase) from sorghum. Plant Physiology 116: 1469–1478.CrossRefGoogle ScholarPubMed
Crombie, HJ (1998) A xyloglucan oligosaccharide-active, trans-glycosylating beta-D-glucosidase from the cotyledons of nasturtium (Tropaeolum majus L) seedlings—purification, properties and characterization of a cDNA clone. Plant Journal 15 (1): 27–38.CrossRefGoogle Scholar
Geerlings, A (2000) Molecular cloning and analysis of strictosidine beta-D-glucosidase, an enzyme in terpenoid indole alkaloid biosynthesis in Catharanthus roseus. Journal of Biological Chemistry 275 (5): 3051–3056.CrossRefGoogle ScholarPubMed
Gusmayer, S (1994) Avenacosidase from oat: purification, sequence-analysis and biochemical characterization of a new member of the BGA family of beta-glucosidases. Plant Molecular Biology 26: 909–921.CrossRefGoogle ScholarPubMed
Jiang, CJ, Wang, ZX and Li, YY (2000) Studies on isolation of general RNA from tea plant [Camellia sinensis (L.) O. Kuntze]. Journal of Tea Science 20 (1): 27.Google Scholar
Ketuat Cairns, JR, Champattanachai, V, Srisomsap, C et al. (2000) Sequence and expression of Thai Rosewood beta-glucosidase/beta-fucosidase, a family 1 glycosyl hydrola glycoprotein. Journal of Biochemistry, (Tokyo) 128: 999–1008.CrossRefGoogle Scholar
Liebig, J and Wohler, F (1837) Formation of the oil of bitter almonds. Annual Chemistry Physics 64: 185–209.Google Scholar
Luo, YP and Tong, QQ (1997) Analysis of the activity of β-glucosidase of the fresh tea shoots of seven cultivars. Journal of Tea Science 17 (Suppl.): 104–107 (in Chinese).Google Scholar
Matsuva, M, Sasaki, J and Murao, S (1995) Studies on β-glucosidase from soybeans that hydrolyze daidzin and genistin, isolation and characterization of an isozyme. Bioscience Biotechnology and Biochemistry 59: 1623–1627.CrossRefGoogle Scholar
Misawa, N and Nakamura, K (1989) Expression and stability of a β-glucosidase gene of Ruminococcus albus in Zymononas mobilis. Agriculture Biology Chemistry 53: 723–727.Google Scholar
Sambrook, J and Russell, WD (2001) Molecular Cloning: A Laboratory Manualr, 3rd ed. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press.Google Scholar
Takeo, T (1981) Production of linalool and geraniol by hydrolytic breakdown of bound forms in disrupted tea shoots. Phytochemistry 120: 2145–2147.CrossRefGoogle Scholar
Tu, YY, Tong, QQ and Luo, YP (1999) The release mechanism of tea aroma: The relationship of β-glucosidase activity and alcoholic aroma Longjing tea processing. Journal of Tea 25 (1): 20–23, 26 (in Chinese).Google Scholar
Wan, XC (1992) Studies on flavor and flavor enzymes of fruit. PhD thesis, Wuxi Light Industrial University (in Chinese).Google Scholar
Wang, Q and Zhao, XH (1994) Catalyze character of β-glucosidase of Aspergillus niger. Research and Development of Natural Products 4: 36–39.Google Scholar
Xia, T and Tong, QQ (1997) Effects of freeze withering on polyphenol oxidase and β-glucosidase activity of tea leaves. Journal of Zheniang Agricultural University 23 (1): 108–110 (in Chinese with English abstract).Google Scholar
Yan, TR and Lin, CL (1997) Purification and characterization of a glucose-tolerant β-glucosidase from Aspergillus niger CCRC 31494. Bioscience Biotechnology and Biochemistry 61: 965–970.CrossRefGoogle ScholarPubMed
Zhang, ZZ (2000) Studies on the major glycodic precusor of green tea. Dissertation of Hunan Agricultural University (in Chinese).Google Scholar