[논문]Discovery and Biochemical Characterization of a Methanol Dehydrogenase From Lysinibacillus xylanilyticus

Lee, Jin-Young; Park, Sung-Hyun; Oh, So-Hyung; Lee, Jin-Ju; Kwon, Kil Koang; Kim, Su-Jin; Choi, Minjeong; Rha, Eugene; Lee, Hyewon; Lee, Dae-Hee; Sung, Bong Hyun; Yeom, Soo-Jin; Lee, Seung-Goo

doi:10.3389/fbioe.2020.00067

[해외논문] Discovery and Biochemical Characterization of a Methanol Dehydrogenase From Lysinibacillus xylanilyticus 원문보기

Frontiers in bioengineering and biotechnology, v.8, 2020년, pp.67 -

Lee, Jin-Young (Synthetic Biology and Bioengineering Research Center, Korea Research Institute of Bioscience and Biotechnology , Daejeon , South Korea) , Park, Sung-Hyun (Synthetic Biology and Bioengineering Research Center, Korea Research Institute of Bioscience and Biotechnology , Daejeon , South Korea) , Oh, So-Hyung (Synthetic Biology and Bioengineering Research Center, Korea Research Institute of Bioscience and Biotechnology , Daejeon , South Korea) , Lee, Jin-Ju (Synthetic Biology and Bioengineering Research Center, Korea Research Institute of Bioscience and Biotechnology , Daejeon , South Korea) , Kwon, Kil Koang (Synthetic Biology and Bioengineering Research Center, Korea Research Institute of Bioscience and Biotechnology , Daejeon , South Korea) , Kim, Su-Jin (Synthetic Biology and Bioengineering Research Center, Korea Research Institute of Bioscience and Biotechnology , Daejeon , South Korea South Kore) , Choi, Minjeong , Rha, Eugene , Lee, Hyewon , Lee, Dae-Hee , Sung, Bong Hyun , Yeom, Soo-Jin , Lee, Seung-Goo

Abstract ▼ AI-Helper

Bioconversion of C1 chemicals such as methane and methanol into higher carbon-chain chemicals has been widely studied. Methanol oxidation catalyzed by methanol dehydrogenase (Mdh) is one of the key steps in methanol utilization in bacterial methylotrophy. In bacteria, few NAD+-dependent Mdhs have been reported that convert methanol to formaldehyde. In this study, an uncharacterized Mdh gene from Lysinibacillus xylanilyticus (Lxmdh) was cloned and expressed in Escherichia coli. The maximum alcohol oxidation activity of the recombinant enzyme was observed at pH 9.5 and 55°C in the presence of 10 mM Mg2+. To improve oxidation activity, rational approach-based, site-directed mutagenesis of 16 residues in the putative active site and NAD+-binding region was performed. The mutations S101V, T141S, and A164F improved the enzyme’s specific activity toward methanol compared to that of the wild-type enzyme. These mutants show a slightly higher turnover rate than that of wild-type, although their KM values were increased compared to that of wild-type. Consequently, according the kinetic results, S101, T141, and A164 positions may related to the catalytic activity in the active site for methanol dehydrogenation. It should be further studied other mutant variants with high activity for methanol. In conclusion, we characterized a new Lxmdh and its variants that may be potentially useful for the development of synthetic methylotrophy in the future.

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참고문헌 (24)

Arfman N. Hektor H. J. Bystrykh L. V. Govorukhina N. I. Dijkhuizen L. Frank J. ( 1997 ). Properties of an NAD(H)-containing methanol dehydrogenase and its activator protein from Bacillus methanolicus . Eur. J. Biochem. 244 426 ？ 433 . 10.1111/j.1432-1033.1997.00426.x 9119008

상세보기
Bradford M. M. ( 1976 ). A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72 248 ？ 254 . 10.1016/0003-2697(76)90527-3 942051

상세보기
Cahn J. K. Baumschlager A. Brinkmann-Chen S. Arnold F. H. ( 2016 ). Mutations in adenine-binding pockets enhance catalytic properties of NAD(P)H-dependent enzymes. Protein Eng. Des. Sel. 29 31 ？ 38 . 10.1093/protein/gzv057 26512129

상세보기
Chen C. T. Chen F. Y. Bogorad I. W. Wu T. Y. Zhang R. Lee A. S. ( 2018 ). Synthetic methanol auxotrophy of Escherichia coli for methanol-dependent growth and production. Metab. Eng. 49 257 ？ 266 . 10.1016/j.ymben.2018.08.010 30172686

상세보기
Chothia C. Lesk A. M. ( 1986 ). The relation between the divergence of sequence and structure in proteins. EMBO J. 5 823 ？ 826 . 10.1002/j.1460-2075.1986.tb04288.x 3709526

상세보기
Hektor H. J. Kloosterman H. Dijkhuizen L. ( 2002 ). Identification of a magnesium-dependent NAD(P)(H)-binding domain in the nicotinoprotein methanol dehydrogenase from Bacillus methanolicus . J. Biol. Chem. 277 46966 ？ 46973 . 10.1074/jbc.m207547200 12351635

상세보기
Krog T. M. Heggeset T. M. Muller J. E. Kupper C. E. Schneider O. Vorholt J. A. ( 2013 ). Methylotrophic Bacillus methanolicus encodes two chromosomal and one plasmid born NAD+ dependent methanol dehydrogenase paralogs with different catalytic and biochemical properties. PLoS One 8 : e59188 . 10.1371/journal.pone.0059188 23527128

상세보기
Laskowski R. A. MacArthur M. W. Moss D. S. Thornton J. M. ( 1993 ). PROCHECK: a program to check the stereochemical quality of protein structures. J. Appl. Crystallogr. 26 283 ？ 291 . 10.1107/s0021889892009944

상세보기
Lesk A. M. Chothia C. ( 1980 ). How different amino acid sequences determine similar protein structures: the structure and evolutionary dynamics of the globins. J. Mol. Biol. 136 225 ？ 230 . 7373651

상세보기
Lesk A. M. Chothia C. ( 1982 ). Evolution of proteins formed by beta-sheets. II. The core of the immunoglobulin domains. J. Mol. Biol. 160 325 ？ 342 . 10.1016/0022-2836(82)90179-6 7175935

상세보기
Marcal D. Rego A. T. Carrondo M. A. Enguita F. J. ( 2009 ). 1,3-Propanediol dehydrogenase from Klebsiella pneumoniae : decameric quaternary structure and possible subunit cooperativity. J. Bacteriol. 191 1143 ？ 1151 . 10.1128/JB.01077-08 19011020

상세보기
Marti-Renom M. A. Stuart A. C. Fiser A. Sanchez R. Melo F. Sali A. ( 2000 ). Comparative protein structure modeling of genes and genomes. Annu. Rev. Biophys. Biomol. Struct. 29 291 ？ 325 . 10.1146/annurev.biophys.29.1.291 10940251

상세보기
Montella C. Bellsolell L. Perez-Luque R. Badia J. Baldoma L. Coll M. ( 2005 ). Crystal structure of an iron-dependent group III dehydrogenase that interconverts L-lactaldehyde and L-1,2-propanediol in Escherichia coli . J. Bacteriol. 187 4957 ？ 4966 . 10.1128/jb.187.14.4957-4966.2005 15995211

상세보기
Moon J. H. Lee H. J. Park S. Y. Song J. M. Park M. Y. Park H. M. ( 2011 ). Structures of iron-dependent alcohol dehydrogenase 2 from Zymomonas mobilis ZM4 with and without NAD+ cofactor. J. Mol. Biol. 407 413 ？ 424 . 10.1016/j.jmb.2011.01.045 21295587

상세보기
Muller J. E. N. Meyer F. Litsanov B. Kiefer P. Potthoff E. Heux S. ( 2015 ). Vorholt, engineering escherichia coli for methanol conversion. Metab. Eng. 28 190 ？ 201 . 10.1016/j.ymben.2014.12.008 25596507

상세보기
Nguyen A. D. Hwang I. Y. Chan J. Y. Lee E. Y. ( 2016 ). Reconstruction of methanol and formate metabolic pathway in non-native host for biosynthesis of chemicals and biofuels. Biotechnol. Bioproces. Eng. 21 477 ？ 482 . 10.1007/s12257-016-0301-7

원문보기 상세보기
Obradors N. Cabiscol E. Aguilar J. Ros J. ( 1998 ). Site-directed mutagenesis studies of the metal-binding center of the iron-dependent propanediol oxidoreductase from Escherichia coli . Eur. J. Biochem. 258 207 ？ 213 . 10.1046/j.1432-1327.1998.2580207.x 9851711

상세보기
Ochsner A. M. Muller J. E. Mora C. A. Vorholt J. A. ( 2014 ). In vitro activation of NAD-dependent alcohol dehydrogenases by nudix hydrolases is more widespread than assumed. FEBS Lett. 588 2993 ？ 2999 . 10.1016/j.febslet.2014.06.008 24928437

상세보기
Read R. J. Brayer G. D. Jurasik L. James M. N. G. ( 1984 ). Critical evaluation of comparative model building of Streptomyces griseus trypsin. Biochemistry 23 6570 ？ 6575 . 10.1021/bi00321a045 6442164

상세보기
Sridhara S. Wu T. T. Chused T. M. Lin E. C. ( 1969 ). Ferrous-activated nicotinamide adenine dinucleotide-linked dehydrogenase from a mutant of Escherichia coli capable of growth on 1, 2-propanediol. J. Bacteriol. 98 87 ？ 95 . 10.1128/jb.98.1.87-95.1969 4306747

상세보기
Trott O. Olson A. J. ( 2010 ). AutoDock vina: improving the speed and accuracy of docking with a new scoring function, efficient optimization, and multithreading. J. Comput. Chem. 31 455 ？ 461 . 10.1002/jcc.21334 19499576

상세보기
Whitaker W. B. Jones J. A. Bennett R. K. Gonzalez J. E. Vernacchio V. R. Collins S. M. ( 2017 ). Engineering the biological conversion of methanol to specialty chemicals in Escherichia coli . Metab. Eng. 39 49 ？ 59 . 10.1016/j.ymben.2016.10.015 27815193

상세보기
Wu T. Y. Chen C. T. Liu J. T. Bogorad I. W. Damoiseaux R. Liao J. C. ( 2016 ). Characterization and evolution of an activator-independent methanol dehydrogenase from Cupriavidus necator N-1. Appl. Microbiol. Biotechnol. 100 4969 ？ 4983 . 10.1007/s00253-016-7320-3 26846745

상세보기
Zhang W. Zhang T. Wu S. Wu M. Xin F. Dong W. ( 2017 ). Guidance for engineering of synthetic methylotrophy based on methanol metabolism in methylotrophy. RSC Adv. 7 4083 ？ 4091 . 10.1039/c6ra27038g

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