Difference between revisions of "Part:BBa K4395022"
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| − | + | SpyTag- UGT-33-SpyCatcher is a fusion protein whose thermostability is increased by the SpyTag-SpyCatcher tagging system while the catalytic activity not affected. | |
| + | UGT-33(UDP-GlucuronosylTransferase33), located in intracellular membrane-bounded organelle, catalyzes the biosynthesis of salidroside from tyrosine. It is a member of the UGT superfamily, which is one of the largest multigene families in Arabidopsis[1]. It has a UDP-glucose-4-O-glucosyltransferase(T8GT) protein coding region. And it is the most active gene among R. rosea Tyrosol-Modifying UGTs. Four UGTs (UGT 17, 29, 32, and 33) were found possessing regio-specific T8GT activity for conversion of tyrosol to salidroside, amongst which UGT-33 displayed the highest activity [2][3]. It is used as the last gene to complete salidroside biosynthesis from tyrosine. | ||
| + | Moreover, UGT-33 exhibits the highest T8GT catalytic efficiency in four UGTs(UGT 2, 3, 29, and 33) with a kcat/KM value of 420.6 s−1 mM−1 and was subsequently referred to as T8GT[2]. | ||
| + | The SpyTag/SpyCatcher system is a split-in-two of the immunoglobulin-like collagen adhesion domain of Streptococcus pyogenes denoted CnaB2. CnaB2 is characterized by an internal isopeptide bond between residue Lys31 and residue Asp117. When split in two, one of which containing Lys31 and another containing Asp117, they associate and spontaneously form the isopeptide bond, thus join together [4] | ||
| + | The composite part SpyTag-UGT-33-SpyCatcher is a fusion protein cyclized by the SpyTag/SpyCatcher system, whose two components are fused on both ends. This cyclization process increases the thermostability of the enzyme, enabling catalysis in an unfavored heated system [5]. | ||
| + | References: | ||
| + | [1] Li, Y. et al. (2001) ‘Phylogenetic analysis of the UDP-glycosyltransferase multigene family of Arabidopsis thaliana’, JOURNAL OF BIOLOGICAL CHEMISTRY, 1 January, pp. 4338–4343. Available at: https://search.ebscohost.com/login.aspx?direct=true&db=edsbl&AN=RN091459152&site=eds-live (Accessed: 27 May 2022). | ||
| + | [2]Michael P. Torrens-Spence;Tomas Pluskal;Fu-Shuang Li;Valentina Carballo;Jing-Ke Weng (2018) ‘Complete Pathway Elucidation and Heterologous Reconstitution of Rhodiola Salidroside Biosynthesis’, 分子植物：英文版 / Molecular Plant, (1), p. 205. Available at: https://search.ebscohost.com/login.aspx?direct=true&db=edscqv&AN=edscqv.674735793&site=eds-live (Accessed: 27 May 2022). | ||
| + | [3] Obesity, Fitness & Wellness Week (2022) ‘Findings from Jiangnan University Yields New Data on Phytochemistry (Biosynthesis and Biotechnological Production of Salidroside From Rhodiola Genus Plants)’, 5 February, p. 101. Available at: https://search.ebscohost.com/login.aspx?direct=true&db=edsggo&AN=edsgcl.690958131&site=eds-live (Accessed: 27 May 2022). | ||
| + | [4] Long Li, Jacob O. Fierer, et al. Structural Analysis and Optimization of the Covalent Association between SpyCatcher and a Peptide Tag J Mol Biol. 2014 January 23; 426(2): 309–317 | ||
| + | [5] Xiao-Bao Suna, Jia-Wen Caoa,b, et al. SpyTag/SpyCatcher molecular cyclization confers protein stability and resilience to aggregation | ||
Latest revision as of 02:56, 4 October 2022
SpyTag- UGT-33-SpyCatcher is a fusion protein whose thermostability is increased by the SpyTag-SpyCatcher tagging system while the catalytic activity not affected. UGT-33(UDP-GlucuronosylTransferase33), located in intracellular membrane-bounded organelle, catalyzes the biosynthesis of salidroside from tyrosine. It is a member of the UGT superfamily, which is one of the largest multigene families in Arabidopsis[1]. It has a UDP-glucose-4-O-glucosyltransferase(T8GT) protein coding region. And it is the most active gene among R. rosea Tyrosol-Modifying UGTs. Four UGTs (UGT 17, 29, 32, and 33) were found possessing regio-specific T8GT activity for conversion of tyrosol to salidroside, amongst which UGT-33 displayed the highest activity [2][3]. It is used as the last gene to complete salidroside biosynthesis from tyrosine. Moreover, UGT-33 exhibits the highest T8GT catalytic efficiency in four UGTs(UGT 2, 3, 29, and 33) with a kcat/KM value of 420.6 s−1 mM−1 and was subsequently referred to as T8GT[2]. The SpyTag/SpyCatcher system is a split-in-two of the immunoglobulin-like collagen adhesion domain of Streptococcus pyogenes denoted CnaB2. CnaB2 is characterized by an internal isopeptide bond between residue Lys31 and residue Asp117. When split in two, one of which containing Lys31 and another containing Asp117, they associate and spontaneously form the isopeptide bond, thus join together [4] The composite part SpyTag-UGT-33-SpyCatcher is a fusion protein cyclized by the SpyTag/SpyCatcher system, whose two components are fused on both ends. This cyclization process increases the thermostability of the enzyme, enabling catalysis in an unfavored heated system [5]. References: [1] Li, Y. et al. (2001) ‘Phylogenetic analysis of the UDP-glycosyltransferase multigene family of Arabidopsis thaliana’, JOURNAL OF BIOLOGICAL CHEMISTRY, 1 January, pp. 4338–4343. Available at: https://search.ebscohost.com/login.aspx?direct=true&db=edsbl&AN=RN091459152&site=eds-live (Accessed: 27 May 2022). [2]Michael P. Torrens-Spence;Tomas Pluskal;Fu-Shuang Li;Valentina Carballo;Jing-Ke Weng (2018) ‘Complete Pathway Elucidation and Heterologous Reconstitution of Rhodiola Salidroside Biosynthesis’, 分子植物：英文版 / Molecular Plant, (1), p. 205. Available at: https://search.ebscohost.com/login.aspx?direct=true&db=edscqv&AN=edscqv.674735793&site=eds-live (Accessed: 27 May 2022). [3] Obesity, Fitness & Wellness Week (2022) ‘Findings from Jiangnan University Yields New Data on Phytochemistry (Biosynthesis and Biotechnological Production of Salidroside From Rhodiola Genus Plants)’, 5 February, p. 101. Available at: https://search.ebscohost.com/login.aspx?direct=true&db=edsggo&AN=edsgcl.690958131&site=eds-live (Accessed: 27 May 2022). [4] Long Li, Jacob O. Fierer, et al. Structural Analysis and Optimization of the Covalent Association between SpyCatcher and a Peptide Tag J Mol Biol. 2014 January 23; 426(2): 309–317 [5] Xiao-Bao Suna, Jia-Wen Caoa,b, et al. SpyTag/SpyCatcher molecular cyclization confers protein stability and resilience to aggregation