Multistep conversion of cresols by phenol hydroxylase and 2,3-dihydroxy-biphenyl 1,2-dioxygenase

Abstract

A multistep conversion system composed of phenol hydroxylase (PH_IND) and 2,3-dihydroxy-biphenyl 1,2-dioxygenase (BphC_LA-4) was used to synthesize methylcatechols and semialdehydes from o- and m-cresol for the first time. Docking studies displayed by PyMOL predicted that cresols and methylcatechols could be theoretically transformed by this multistep conversion system. High performance liquid chromatography mass spectrometry (HPLC-MS) analysis also indicated that the products formed from multistep conversion were the corresponding 3-methylcatechol, 4-methylcatechol, 2-hydroxy-3-methyl-6-oxohexa-2,4-dienoic acid (2-hydroxy-3-methyl-ODA) and 2-hydroxy-5-methyl-6-oxohexa-2,4-dienoic acid (2-hydroxy-5-methyl-ODA). The optimal cell concentrations of the recombinant E. coli strain BL21 (DE3) expressing phenol hydroxylase (PH_IND) and 2,3-dihydroxy-biphenyl 1,2-dioxygenase (BphC_LA-4) and pH for the multistep conversion of o- and m-cresol were 4.0 (g·L^-1 cell dry weight) and pH 8.0, respectively. For the first step conversion, the formation rate of 3-methylcatechol (0.29 μmol·L^-1·min^-1·mg^-1 cell dry weight) from o-cresol was similarly with that of methylcatechols (0.28 μmol·L^-1·min^-1·mg^-1 cell dry weight) from m-cresol by strain PH_IND. For the second step conversion, strain BphC_LA-4 showed higher formation rate (0.83 μmol·L^-1·min^-1·mg^-1 cell dry weight) for 2-hydroxy-3-methyl-ODA and 2-hydroxy-5-methyl-ODA from m-cresol, which was 1.1-fold higher than that for 2-hydroxy-3-methyl-ODA (0.77 μmol·L^-1·min^-1·mg^-1cell dry weight) from o-cresol. The present study suggested the potential application of the multistep conversion system for the production of chemical synthons and high-value products.

Keywords

multistep conversion / cresols / phenol hydroxylase / 2 / 3-dihydroxybiphenyl 1 / 2-dioxygenase / methylcatechols

References

[1] MüllerJ A, GalushkoA S, KapplerA, SchinkB. Anaerobic degradation of m-cresol by Desulfobacterium cetonicum is initiated by formation of 3-hydroxybenzylsuccinate. Archives of Microbiology, 1999, 172(5): 287-294

[2] LondryK L, FedorakP M, SuflitaJ M. Anaerobic degradation of m-Cresol by a sulfate-reducing bacterium. Applied and Environmental Microbiology, 1997, 63(8): 3170-3175

[3] TallurP N, MegadiV B, KamanavalliC M, NinnekarH Z. Biodegradation of p-cresol by Bacillus sp. strain PHN 1. Current Microbiology, 2006, 53(6): 529-533

[4] KnackmussH J. Biochemistry and practical implications of organohalide degradation. In: KlugM J, ReddyC A, eds. Current Perspectives in Microbial. Washington, D C: American Society for Microbiology, 1984, 687-693

[5] DagleyS, GibsonD T. The bacterial degradation of catechol. The Biochemical Journal, 1965, 95(2): 466-474

[6] CoulombelL, NolanL C, NikodinovicJ, DoyleE M, O’ConnorK E. Biotransformation of 4-halophenols to 4-halocatechols using Escherichia coli expressing 4-hydroxyphenylacetate 3-hydroxylase. Applied Microbiology and Biotechnology, 2011, 89(6): 1867-1875

[7] QuY Y, ShiS N, QiaoM, KongC L, ZhouH, ZhangX W, ZhouJ T. Multistep conversion of para-substituted phenols by phenol hydroxylase and 2,3-dihydroxy-biphenyl 1,2-dioxygenase. Applied Biochemistry and Biotechnology, 2013, 169(7): 2064-2075.

[8] TaoY, FishmanA, BentleyW E, WoodT K. Oxidation of benzene to phenol, catechol, and 1,2,3-trihydroxybenzene by toluene 4-monooxygenase of Pseudomonas mendocina KR1 and toluene 3-monooxygenase of Ralstonia pickettii PKO1. Applied and Environmental Microbiology, 2004, 70(7): 3814-3820

[9] RobertsS J, MorrisJ C, DobsonR C J, GerrardJ A. The preparation of (S)-aspartate semi-aldehyde appropriate for use in biochemical studies. Bioorganic & Medicinal Chemistry Letters, 2003, 13(2): 265-267

[10] ParalesR E, BruceN C, SchmidA, WackettL P. Biodegradation, biotransformation, and biocatalysis (b3). Applied and Environmental Microbiology, 2002, 68(10): 4699-4709

[11] GarikipatiS V, McIverA M, PeeplesT L. Whole-cell biocatalysis for 1-naphthol production in liquid-liquid biphasic systems. Applied and Environmental Microbiology, 2009, 75(20): 6545-6552

[12] PollardD J, WoodleyJ M. Biocatalysis for pharmaceutical intermediates: the future is now. Trends in Biotechnology, 2007, 25(2): 66-73

[13] SchmidA, DordickJ S, HauerB, KienerA, WubboltsM, WitholtB. Industrial biocatalysis today and tomorrow. Nature, 2001, 409(6817): 258-268

[14] AzeradR. Editorial overview: better enzyme for green chemistry. Current Opinion in Biotechnology, 2001, 12(6): 533-534

[15] StraathofA J J, PankeS, SchmidA. The production of fine chemicals by biotransformations. Current Opinion in Biotechnology, 2002, 13(6): 548-556

[16] SelinheimoE, GasparettiC, MattinenM L, SteffensenC L, BuchertJ, KruusK. Comparison of substrate specificity of tyrosinases from Trichoderma reesei and Agaricus bisporus. Enzyme and Microbial Technology, 2009, 44(1): 1-10

[17] SazinskyM H, DuntenP W, McCormickM S, DiDonatoA, LippardS J. X-ray structure of a hydroxylase-regulatory protein complex from a hydrocarbon-oxidizing multicomponent monooxygenase, Pseudomonas sp. OX1 phenol hydroxylase. Biochemistry, 2006, 45(51): 15392-15404

[18] EltisL D, HofmannB, HechtH J, LünsdorfH, TimmisK N. Purification and crystallization of 2,3-dihydroxybiphenyl 1,2-dioxygenase. The Journal of Biological Chemistry, 1993, 268(4): 2727-2732

[19] WuZ L, PodustL M, GuengerichF P. Expansion of substrate specificity of cytochrome P450 2A6 by random and site-directed mutagenesis. The Journal of Biological Chemistry, 2005, 280(49): 41090-41100

[20] MorrisG M, Lim-WilbyM. Molecular docking. Methods in Molecular Biology (Clifton, N.J.), 2008, 443(1064-3745): 365-382

[21] MaF, ShiS N, SunT H, LiA, ZhouJ T, QuY Y. Biotransformation of benzene and toluene to catechols by phenol hydroxylase from Arthrobacter sp. W1. Applied Microbiology and Biotechnology, 2013, 97(11): 5097-5103

[22] CarliellC M, BarclayS J, NaidooN, BuckleyC A, MulhollandD A, SeniorE. Microbial decolorization of a reactive azo dye under anaerobic conditions. Water SA, 1995, 21(1): 61-69

RIGHTS & PERMISSIONS

Higher Education Press and Springer-Verlag Berlin Heidelberg