- Email: [email protected]
water mixtures catalyzed by vanadium bromoperoxidase from Corallina officinalis
Abstracts / Journal of Biotechnology 136S (2008) S356–S401
S369
V3-P-033
V3-P-035
HPLC-PAD–ESI/MS/MS: Tool for investigation of kinetics of enzymatic hydrolysis of mono-caffeoylquinic acids from tobacco waste
Epoxidation of cyclohexene in t-butyl alcohol/water mixtures catalyzed by vanadium bromoperoxidase from Corallina officinalis
Jun Wang 1 , Dingqiang Lu 1,2,∗ , Hongqun Qiao 2 , Xiuquan Ling 1 , Hui Zhao 1 , Pingkai Ouyang 1
Biaoming Zhang 1,2 , Peichun Wu 1 , Wei Zhang 1,3,∗
1
College of Life Science and Pharmaceutical Engineering, Nanjing University of Technology, Nanjing 210009, PR China 2 Jiangsu Provincial Institute of Materia Medica, Nanjing 210009, PR China E-mail address: [email protected] (D. Lu).
The hydrolysis products of mono-caffeoylquinic acids (mono-CQAs) are caffeic acid (CA) and d-(−)-quinic acid (QA) which have been shown to exhibit various biological and pharmacological properties (Arthurs et al., 2008). In order to investigate accurately the kinetics of enzymatic hydrolysis of mono-CQAs from tobacco waste extract (TWE), ion trap HPLC-PAD–ESI/MS/MS was used as a qualitative and quantitative tool in the analysis of contents variation of chlorogenic acid (5-CQA), neochlorogenic acid (3-CQA) and cryptochlorogenic acid (4-CQA) in the reaction system. Lipase was used to catalyze the hydrolysis of the above compounds to obtain caffeic acid and d-(−)-quinic acid (Yoshimoto et al., 2005), and LC–MS/MS (Clifford et al., 2008) was used to characterize cinnamoylamino acid conjugates and to discriminate between individual isomers of mono-acyl chlorogenic acids. The substrates and products were separated on a Alltima C18 column (250 mm × 4.6 mm i.d.; 5 m) with a mobile phase consisting acetonitrile:ammonium acetate buffer (pH 4.5) (3:97, v/v), at a flow rate of 1.0 mL min−1 , detected at 327 nm, and four regression equations showed good linear relationships (r2 > 0.999) between the peak area of each marker and concentration. Treatment of the kinetic data was carried out by non-linear regression analysis. The results showed that the lipase was found to exert low specificity to individual mono-CQAs in the hydrolysed TWE solution, and the developed assay method was rapid, accurate, reliable and be readily utilized as an analysis method for elucidating the kinetics of enzymatic hydrolysis of mono-CQAs.
1
Marine Bioproducts Engineering Group, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, China 2 Graduate School of Chinese Academy of Sciences, Beijing 100039, China 3 Department of Medical Biotechnology, School of Medicine, Flinders University, Bedford Park, Adelaide, SA 5042, Australia E-mail address: [email protected] (W. Zhang). Current interests in catalytic oxidative transformations in chemical industry are to replace oxidation catalysts that consist of heavy metals, and to develop catalysts with high chemo-, regio- or enantioselectivities (Valery, 2003). Haloperoxidases are potentially suitable biocatalysts to meet these goals (Valery, 2003). In recent years, there is a substantial interest on vanadium bromoperoxidase (VBPO) due to its excellent thermostability, operation stability and broad spectra of substrates such as methyl phenyl ether, 1-methoxylnaphthalenethiophene, indene, organic sulfide, indole terpenes (Valery, 2003; Itoh et al., 1988). In our previous studies we have investigated the epoxidation of cyclohexene to form 1,2cyclohexane catalyzed by VBPO isolated from the marine red algae Corallina officinalis with a yield of 2.8 g l−1 h−1 and 82% selectivity (Yu et al., 2007). The epoxidation was performed in a water-organic biphasic solvent system and the solubility of the cyclohexene is too low in water. In this work, t-butyl alcohol was used as a co-solvent in order to enhance the solubility of cyclohexene in water. A final yield of 3.2 g l−1 h−1 of 1,2-cyclohexane with 96% selectivity was achieved. The reaction was completed within 4 h that is shorter than that without co-solvent (6 h). The results indicated that tbutyl alcohol/water (2%, v/v) was an excellent co-solvent system to improve the bio-oxidation efficiency of cyclohexene by VBPO, with further optimization required. References
References Arthurs, C.L., Lingley, K.F., Piacenti, M., Stratford, I.J., Tatic, T., Whitehead, R.C., Wind, N.S., 2008. (−)-Quinic acid: a versatile precursor for the synthesis of analogues of 2-crotonyloxymethyl-(4R,5R,6R)-4,5,6-trihydroxycyclohex-2-enone (COTC) which possess anti-tumour properties. Tetrahedron Lett. 49, 2410–2413. Clifford, M.N., Kirkpatrick, J., Kuhnert, N., Roozendaal, H., Salgado, P.R., 2008. LC–MSn analysis of the cis isomers of chlorogenic acids. Food Chem. 106, 379–385. Yoshimoto, M., Kurata-Azuma, R., Fujii, M., Hou, D.X., Ikeda, K., Yoshidome, T., Osako, M., 2005. Enzymatic production of caffeic acid by koji from plant resources containing caffeoylquinic acid derivatives. Biosci. Biotechnol. Biochem. 69, 1777–1781.
Itoh, N., Hasan, Q., Izumi, Y., Yamada, H., 1988. Substrate specificity, regiospecificity and stereospecificity of halogenation reactions catalyzed by non-heme-type bromoperoxidase of Corallina pilulifera. Eur. J. Biochem. 172, 477–484. Valery, M.D., 2003. Oxidation, epoxidation and sulfoxidation reactions catalysed by haloperoxidases. Tetrahedron 59, 4701–4720. Yu, Y., Jin, Y., Wu, P.C., Zhang, W., 2007. Epoxidation of cyclohexene catalyzed by the vanadium bromoperoxidase from Corallina officinalis. Chin. J. Catal. 28, 915–918.
doi:10.1016/j.jbiotec.2008.07.849 V3-P-036
doi:10.1016/j.jbiotec.2008.07.848 Biocatalytic production of glycolic acid from glycolonitrile with a new bacterial isolate of Alcaligenes sp. ECU0401 Yu-Cai He ∗ , Jian-He Xu ∗ Laboratory of Biocatalysis and Bioprocessing, State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China E-mail addresses: [email protected] (Y.-C. He), [email protected] (J.-H. Xu). ␣-Hydroxy carboxylic acids are an important class of compounds (He et al., 2007, 2008). Among them, glycolic acid (HOCH2 COOH) is known to be the simplest kind of ␣-hydroxy carboxylic compounds. Its properties make it ideal for a broad spectrum of industrial and
S369
V3-P-033
V3-P-035
HPLC-PAD–ESI/MS/MS: Tool for investigation of kinetics of enzymatic hydrolysis of mono-caffeoylquinic acids from tobacco waste
Epoxidation of cyclohexene in t-butyl alcohol/water mixtures catalyzed by vanadium bromoperoxidase from Corallina officinalis
Jun Wang 1 , Dingqiang Lu 1,2,∗ , Hongqun Qiao 2 , Xiuquan Ling 1 , Hui Zhao 1 , Pingkai Ouyang 1
Biaoming Zhang 1,2 , Peichun Wu 1 , Wei Zhang 1,3,∗
1
College of Life Science and Pharmaceutical Engineering, Nanjing University of Technology, Nanjing 210009, PR China 2 Jiangsu Provincial Institute of Materia Medica, Nanjing 210009, PR China E-mail address: [email protected] (D. Lu).
The hydrolysis products of mono-caffeoylquinic acids (mono-CQAs) are caffeic acid (CA) and d-(−)-quinic acid (QA) which have been shown to exhibit various biological and pharmacological properties (Arthurs et al., 2008). In order to investigate accurately the kinetics of enzymatic hydrolysis of mono-CQAs from tobacco waste extract (TWE), ion trap HPLC-PAD–ESI/MS/MS was used as a qualitative and quantitative tool in the analysis of contents variation of chlorogenic acid (5-CQA), neochlorogenic acid (3-CQA) and cryptochlorogenic acid (4-CQA) in the reaction system. Lipase was used to catalyze the hydrolysis of the above compounds to obtain caffeic acid and d-(−)-quinic acid (Yoshimoto et al., 2005), and LC–MS/MS (Clifford et al., 2008) was used to characterize cinnamoylamino acid conjugates and to discriminate between individual isomers of mono-acyl chlorogenic acids. The substrates and products were separated on a Alltima C18 column (250 mm × 4.6 mm i.d.; 5 m) with a mobile phase consisting acetonitrile:ammonium acetate buffer (pH 4.5) (3:97, v/v), at a flow rate of 1.0 mL min−1 , detected at 327 nm, and four regression equations showed good linear relationships (r2 > 0.999) between the peak area of each marker and concentration. Treatment of the kinetic data was carried out by non-linear regression analysis. The results showed that the lipase was found to exert low specificity to individual mono-CQAs in the hydrolysed TWE solution, and the developed assay method was rapid, accurate, reliable and be readily utilized as an analysis method for elucidating the kinetics of enzymatic hydrolysis of mono-CQAs.
1
Marine Bioproducts Engineering Group, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, China 2 Graduate School of Chinese Academy of Sciences, Beijing 100039, China 3 Department of Medical Biotechnology, School of Medicine, Flinders University, Bedford Park, Adelaide, SA 5042, Australia E-mail address: [email protected] (W. Zhang). Current interests in catalytic oxidative transformations in chemical industry are to replace oxidation catalysts that consist of heavy metals, and to develop catalysts with high chemo-, regio- or enantioselectivities (Valery, 2003). Haloperoxidases are potentially suitable biocatalysts to meet these goals (Valery, 2003). In recent years, there is a substantial interest on vanadium bromoperoxidase (VBPO) due to its excellent thermostability, operation stability and broad spectra of substrates such as methyl phenyl ether, 1-methoxylnaphthalenethiophene, indene, organic sulfide, indole terpenes (Valery, 2003; Itoh et al., 1988). In our previous studies we have investigated the epoxidation of cyclohexene to form 1,2cyclohexane catalyzed by VBPO isolated from the marine red algae Corallina officinalis with a yield of 2.8 g l−1 h−1 and 82% selectivity (Yu et al., 2007). The epoxidation was performed in a water-organic biphasic solvent system and the solubility of the cyclohexene is too low in water. In this work, t-butyl alcohol was used as a co-solvent in order to enhance the solubility of cyclohexene in water. A final yield of 3.2 g l−1 h−1 of 1,2-cyclohexane with 96% selectivity was achieved. The reaction was completed within 4 h that is shorter than that without co-solvent (6 h). The results indicated that tbutyl alcohol/water (2%, v/v) was an excellent co-solvent system to improve the bio-oxidation efficiency of cyclohexene by VBPO, with further optimization required. References
References Arthurs, C.L., Lingley, K.F., Piacenti, M., Stratford, I.J., Tatic, T., Whitehead, R.C., Wind, N.S., 2008. (−)-Quinic acid: a versatile precursor for the synthesis of analogues of 2-crotonyloxymethyl-(4R,5R,6R)-4,5,6-trihydroxycyclohex-2-enone (COTC) which possess anti-tumour properties. Tetrahedron Lett. 49, 2410–2413. Clifford, M.N., Kirkpatrick, J., Kuhnert, N., Roozendaal, H., Salgado, P.R., 2008. LC–MSn analysis of the cis isomers of chlorogenic acids. Food Chem. 106, 379–385. Yoshimoto, M., Kurata-Azuma, R., Fujii, M., Hou, D.X., Ikeda, K., Yoshidome, T., Osako, M., 2005. Enzymatic production of caffeic acid by koji from plant resources containing caffeoylquinic acid derivatives. Biosci. Biotechnol. Biochem. 69, 1777–1781.
Itoh, N., Hasan, Q., Izumi, Y., Yamada, H., 1988. Substrate specificity, regiospecificity and stereospecificity of halogenation reactions catalyzed by non-heme-type bromoperoxidase of Corallina pilulifera. Eur. J. Biochem. 172, 477–484. Valery, M.D., 2003. Oxidation, epoxidation and sulfoxidation reactions catalysed by haloperoxidases. Tetrahedron 59, 4701–4720. Yu, Y., Jin, Y., Wu, P.C., Zhang, W., 2007. Epoxidation of cyclohexene catalyzed by the vanadium bromoperoxidase from Corallina officinalis. Chin. J. Catal. 28, 915–918.
doi:10.1016/j.jbiotec.2008.07.849 V3-P-036
doi:10.1016/j.jbiotec.2008.07.848 Biocatalytic production of glycolic acid from glycolonitrile with a new bacterial isolate of Alcaligenes sp. ECU0401 Yu-Cai He ∗ , Jian-He Xu ∗ Laboratory of Biocatalysis and Bioprocessing, State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China E-mail addresses: [email protected] (Y.-C. He), [email protected] (J.-H. Xu). ␣-Hydroxy carboxylic acids are an important class of compounds (He et al., 2007, 2008). Among them, glycolic acid (HOCH2 COOH) is known to be the simplest kind of ␣-hydroxy carboxylic compounds. Its properties make it ideal for a broad spectrum of industrial and