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http://dx.doi.org/10.4014/mbl.1905.05008

Application of Solanum lycopersicum Glucose-6-phosphate Dehydrogenase to NADPH-generating System for Cytochrome P450 Reactions  

Park, Chan Mi (School of Biological Sciences and Technology, Chonnam National University)
Jeong, Heon (School of Biological Sciences and Technology, Chonnam National University)
Ma, Sang Hoon (School of Biological Sciences and Technology, Chonnam National University)
Kim, Hyun Min (School of Biological Sciences and Technology, Chonnam National University)
Joung, Young Hee (School of Biological Sciences and Technology, Chonnam National University)
Yun, Chul-Ho (School of Biological Sciences and Technology, Chonnam National University)
Publication Information
Microbiology and Biotechnology Letters / v.47, no.4, 2019 , pp. 536-545 More about this Journal
Abstract
Cytochrome P450 (P450 or CYP) is involved in the metabolism of endogenous and exogenous compounds in most organisms. P450s have great potential as biocatalysts in the pharmaceutical and fine chemical industries because they catalyze diverse oxidative reactions using a wide range of substrates. The high-cost nicotinamide cofactor, NADPH, is essential for P450 reactions. Glucose-6-phosphate dehydrogenase (G6PDH) has been commonly used in NADPH-generating systems (NGSs) to provide NADPH for P450 reactions. Currently, only two G6PDHs from Leuconostoc mesenteroides and Saccharomyces cerevisiae can be obtained commercially. To supply high-cost G6PDH cost-effectively, we cloned the cytosolic G6PDH gene of Solanum lycopersicum (tomato) with 6xHis tag, expressed it in Escherichia coli, and purified the recombinant G6PDH (His-G6PDH) using affinity chromatography. In addition, enzymatic properties of His-G6PDH were investigated, and the His-G6PDH-coupled NGS was optimized for P450 reactions. His-G6PDH supported CYP102A1-catalyzed hydroxylation of omeprazole and testosterone by NADPH generation. This result suggests that tomato His-G6PDH could be a cost-effective enzyme source for NGSs for P450-catalyzed reactions as well as other NADPH-requiring reactions.
Keywords
Cytochrome P450; tomato glucose-6-phosphate dehydrogenase; heterologous expression; NADPH-generating system;
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1 Hall TA. 1999. BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. http://www.mbio.ncsu.edu/BioEdit/bioedit.html. Accessed May 8, 2019.
2 Castiglia D, Cardi M, Landi S, Cafasso D, Esposito S. 2015. Expression and characterization of a cytosolic glucose 6 phosphate dehydrogenase isoform from barley (Hordeum vulgare) roots. Protein Expr. Purif. 112: 8-14.   DOI
3 Cardi M, Chibani K, Castiglia D, Cafasso D, Pizzo E, Rouhier N, et al. 2013. Overexpression, purification and enzymatic characterization of a recombinant plastidial glucose-6-phosphate dehydrogenase from barley (Hordeum vulgare cv. Nure) roots. Plant Physiol. Biochem. 73: 266-273.   DOI
4 Yang Y, Fu Z, Su Y, Zhang X, Li G, Guo J, et al. 2014. A cytosolic glucose-6-phosphate dehydrogenase gene, ScG6PDH, plays a positive role in response to various abiotic stresses in sugarcane. Sci. Rep. 4: 7090.   DOI
5 Cardi M, Castiglia D, Ferrara M, Guerriero G, Chiurazzi M, Esposito S. 2015. The effects of salt stress cause a diversion of basal metabolism in barley roots: possible different roles for glucose-6-phosphate dehydrogenase isoforms. Plant Physiol. Biochem. 86: 44-54.   DOI
6 Liu J, Wang X, Hu Y, Hu W, Bi Y. 2013. Glucose-6-phosphate dehydrogenase plays a pivotal role in tolerance to drought stress in soybean roots. Plant Cell Rep. 32: 415-429.   DOI
7 Wang H, Yang L, Li Y, Hou J, Huang J, Liang W. 2016. Involvement of ABA- and $H_2O_2$-dependent cytosolic glucose-6-phosphate dehydrogenase in maintaining redox homeostasis in soybean roots under drought stress. Plant Physiol. Biochem. 107: 126-136.   DOI
8 Landi S, Nurcato R, De Lillo A, Lentini M, Grillo S, Esposito S. 2016. Glucose-6-phosphate dehydrogenase plays a central role in the response of tomato (Solanum lycopersicum) plants to short and long-term drought. Plant Physiol. Biochem. 105: 79-89.   DOI
9 Kim DH, Kim KH, Kim DH, Liu KH, Jung HC, Pan JG, et al. 2008. Generation of human metabolites of 7-ethoxycoumarin by bacterial cytochrome P450 BM3. Drug Metab. Dispos. 36: 2166-2170.   DOI
10 Omura T, Sato R. 1964. The carbon monoxide-binding pigment of liver microsomes. II. solubilization, purification, and properties. J. Biol. Chem. 239: 2379-2385.   DOI
11 Ryu SH, Park BY, Kim SY, Park SH, Jung HJ, Park M, et al. 2014. Regioselective hydroxylation of omeprazole enantiomers by bacterial CYP102A1 mutants. Drug Metab. Dispos. 42: 1493-1497.   DOI
12 Vottero E, Rea V, Lastdrager J, Honing M, Vermeulen NP, Commandeur JN. 2011. Role of residue 87 in substrate selectivity and regioselectivity of drug-metabolizing cytochrome P450 CYP102A1 M11. J. Biol. Inorg. Chem. 16: 899-912.   DOI
13 NCBI. Available from https://www.ncbi.nlm.nih.gov. Accessed May 8, 2019.
14 UniProtKB. Available from https://www.uniprot.org/. Accessed May 8, 2019.
15 Multiple Sequence Alignment by CLUSTALW. Available from https://www.genome.jp/tools-bin/clustalw. Accessed May 8, 2019.
16 Basic Local Alignment Search Tool. Available from https://blast.ncbi.nlm.nih.gov/Blast.cgi. Accessed May 8, 2019.
17 Translate Tool. Available from https://web.expasy.org/translate/. Accessed May 8, 2019.
18 Guengerich FP. 2008. Cytochrome P450 and chemical toxicology. Chem. Res. Toxicol. 21: 70-83.   DOI
19 Tang W, Stearns RA. 2001. Heterotropic cooperativity of cytochrome P450 3A4 and potential drug-drug interactions. Curr. Drug Metab. 2: 185-198.   DOI
20 Guengerich FP. 2001. Common and uncommon cytochrome P450 reactions related to metabolism and chemical toxicity. Chem. Res. Toxicol. 14: 611-650.   DOI
21 Guengerich FP, Munro AW. 2013. Unusual cytochrome P450 enzymes and reactions. J. Biol. Chem. 288: 17065-17073.   DOI
22 Munro AW, Daff S, Coggins JR, Lindsay JG, Chapman SK. 1996. Probing electron transfer in flavocytochrome P-450 BM3 and its component domains. Eur. J. Biochem. 239: 403-409.   DOI
23 Yun CH, Kim KH, Kim DH, Jung HC, Pan JG. 2007. The bacterial P450 BM3: a prototype for a biocatalyst with human P450 activities. Trends Biotechnol. 25: 289-298.   DOI
24 Kang JY, Ryu SH, Park SH, Cha GS, Kim DH, Kim KH, et al. 2014. Chimeric cytochromes P450 engineered by domain swapping and random mutagenesis for producing human metabolites of drugs. Biotechnol. Bioeng. 111: 1313-1322.   DOI
25 Lussenburg BMA, Babel LC, Vermeulen NPE, Commandeur JNM. 2005. Evaluation of alkoxyresorufins as fluorescent substrates for cytochrome P450 BM3 and site-directed mutants. Anal. Biochem. 341: 148-155.   DOI
26 Xu Z, Jing K, Liu Y, Cen P. 2007. High-level expression of recombinant glucose dehydrogenase and its application in NADPH regeneration. J. Ind. Microbiol. Biotechnol. 34: 83-90.   DOI
27 Uppada V, Bhaduri S, Noronha SB. 2014. Cofactor regenerationan important aspect of biocatalysis. Curr. Sci. 106: 946-957.
28 Kruger NJ, von Schaewen A. 2003. The oxidative pentose phosphate pathway: structure and organisation. Curr. Opin. Plant Biol. 6: 236-246.   DOI
29 Schnarrenber C, Oeser A, Tolbert NE. 1973. Two isoenzymes each of glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase in spinach leaves. Arch. Biochem. Biophys. 154: 438-448.   DOI
30 von Schaewen A, Langenkamper G, Graeve K, Wenderoth I, Scheibe R. 1995. Molecular characterization of the plastidic glucose- 6-phosphate dehydrogenase from potato in comparison to its cytosolic counterpart. Plant Physiol. 109: 1327-1335.   DOI
31 Gnanasekaran T, Vavitsas K, Andersen-Ranberg J, Nielsen AZ, Olsen CE, Hamberger B, et al. 2015. Heterologous expression of the isopimaric acid pathway in Nicotiana benthamiana and the effect of N-terminal modifications of the involved cytochrome P450 enzyme. J. Biol. Eng. 9: 24-24.   DOI
32 Sheludko YV, Gerasymenko IM, Warzecha H. 2018. Transient expression of human cytochrome P450s 2D6 and 3A4 in Nicotiana benthamiana provides a Possibility for Rapid Substrate Testing and Production of novel compounds. Biotechnol. J. 13: e1700696.
33 Schadt S, Bister B, Chowdhury SK, Funk C, Hop CECA, Humphreys WG, et al. 2018. A Decade in the MIST: Learnings from Investigations of Drug Metabolites in Drug Development under the "Metabolites in Safety Testing" Regulatory Guidance. Drug Metab. Dispos. 46: 865-878.   DOI
34 Furge LL, Guengerich FP. 2006. Cytochrome P450 enzymes in drug metabolism and chemical toxicology: An introduction. Biochem. Mol. Biol. Educ. 34: 66-74.   DOI
35 Urlacher VB, Eiben S. 2006. Cytochrome P450 monooxygenases: perspectives for synthetic application. Trends Biotechnol. 24: 324-330.   DOI
36 Schewe H, Kaup BA, Schrader J. 2008. Improvement of P450 (BM-3) whole-cell biocatalysis by integrating heterologous cofactor regeneration combining glucose facilitator and dehydrogenase in E. coli. Appl. Microbiol. Biotechnol. 78: 55-65.   DOI
37 Siriphongphaew A, Pisnupong P, Wongkongkatep J, Inprakhon P, Vangnai AS, Honda K, et al. 2012. Development of a whole-cell biocatalyst co-expressing P450 monooxygenase and glucose dehydrogenase for synthesis of epoxyhexane. Appl. Microbiol. Biotechnol. 95: 357-367.   DOI
38 Cirino PC, Arnold FH. 2003. A self-sufficient peroxide-driven hydroxylation biocatalyst. Angew. Chem. Int. Ed. Engl. 42: 3299-3301.   DOI
39 Fantuzzi A, Fairhead M, Gilardi G. 2004. Direct electrochemistry of immobilized human cytochrome P450 2E1. J. Am. Chem. Soc. 126: 5040-5041.   DOI
40 Bian M, Li S, Wei H, Huang S, Zhou F, Zhu Y, et al. 2018. Heteroexpression and biochemical characterization of a glucose-6-phosphate dehydrogenase from oleaginous yeast Yarrowia lipolytica. Protein Expr. Purif. 148: 1-8.   DOI
41 Levy HR, Daouk GH. 1979. Simultaneous analysis of NAD- and NADP-linked activities of dual nucleotide-specific dehydrogenases. Application to Leuconostoc mesenteroides glucose-6-phosphate dehydrogenase. J. Biol. Chem. 254: 4843-4847.   DOI
42 Adediran SA, Gbadegesin MR. 1995. Kinetics of the reaction of baker's yeast glucose-6-phosphate dehydrogenase with 5, 5'-dithiobis (2-nitrobenzoic acid). Arch. Biochem. Biophys. 322: 39-42.   DOI
43 Cave CRJ, Cockshull KE, Adams SR. 2001. Effect of temperature on the growth and development of tomato fruits. Ann. Bot. 88: 869-877.   DOI
44 Demoss RD, Gunsalus IC, Bard RC. 1953. A glucose-6-phosphate dehydrogenase in Leuconostoc mesenteroides. J. Bacteriol. 66: 10-16.   DOI
45 Glaser L, Brown DH. 1955. Purification and properties of d-glucose- 6-phosphate dehydrogenase. J. Biol. Chem. 216: 67-79.   DOI
46 Jeon H, Durairaj P, Lee D, Ahsan MM, Yun H. 2016. Improved NADPH regeneration for fungal cytochrome P450 monooxygenase by co-expressing bacterial glucose dehydrogenase in restingcell biotransformation of recombinant yeast. J. Microbiol. Biotechnol. 26: 2076-2086.   DOI
47 ProtParam tool. Available from https://web.expasy.org/protparam/. Accessed May 8, 2019.
48 Corpas FJ, Barroso JB, Sandalio LM, Distefano S, Palma JM, Lupianez JA, et al. 1998. A dehydrogenase-mediated recycling system of NADPH in plant peroxisomes. Biochem J. 330(Pt 2): 777-784.   DOI
49 Wakao S, Benning C. 2005. Genome-wide analysis of glucose-6- phosphate dehydrogenases in Arabidopsis. Plant J. 41: 243-256.   DOI
50 Esposito S, Carfagna S, Massaro G, Vona V, Di Martino Rigano V. 2001. Glucose-6-phosphate dehydrogenase in barley roots: kinetic properties and localisation of the isoforms. Planta 212: 627-634.   DOI
51 Graeve K, von Schaewen A, Scheibe R. 1994. Purification, characterization, and cDNA sequence of glucose-6-phosphate dehydrogenase from potato (Solanum tuberosum L.). Plant J. 5: 353-361.   DOI