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http://dx.doi.org/10.4014/jmb.2103.03030

NADP+-Dependent Dehydrogenase SCO3486 and Cycloisomerase SCO3480: Key Enzymes for 3,6-Anhydro-ʟ-Galactose Catabolism in Streptomyces coelicolor A3(2)  

Tsevelkhorloo, Maral (Department of Biosciences and Bioinformatics, Myongji University)
Kim, Sang Hoon (Department of Animal Resources Science, Dankook University)
Kang, Dae-Kyung (Department of Animal Resources Science, Dankook University)
Lee, Chang-Ro (Department of Biosciences and Bioinformatics, Myongji University)
Hong, Soon-Kwang (Department of Biosciences and Bioinformatics, Myongji University)
Publication Information
Journal of Microbiology and Biotechnology / v.31, no.5, 2021 , pp. 756-763 More about this Journal
Abstract
Agarose is a linear polysaccharide composed of ᴅ-galactose and 3,6-anhydro-ʟ-galactose (AHG). It is a major component of the red algal cell wall and is gaining attention as an abundant marine biomass. However, the inability to ferment AHG is considered an obstacle in the large-scale use of agarose and could be addressed by understanding AHG catabolism in agarolytic microorganisms. Since AHG catabolism was uniquely confirmed in Vibrio sp. EJY3, a gram-negative marine bacterial species, we investigated AHG metabolism in Streptomyces coelicolor A3(2), an agarolytic gram-positive soil bacterium. Based on genomic data, the SCO3486 protein (492 amino acids) and the SCO3480 protein (361 amino acids) of S. coelicolor A3(2) showed identity with H2IFE7.1 (40% identity) encoding AHG dehydrogenase and H2IFX0.1 (42% identity) encoding 3,6-anhydro-ʟ-galactonate cycloisomerase, respectively, which are involved in the initial catabolism of AHG in Vibrio sp. EJY3. Thin layer chromatography and mass spectrometry of the bioconversion products catalyzed by recombinant SCO3486 and SCO3480 proteins, revealed that SCO3486 is an AHG dehydrogenase that oxidizes AHG to 3,6-anhydro-ʟ-galactonate, and SCO3480 is a 3,6-anhydro-ʟ-galactonate cycloisomerase that converts 3,6-anhydro-ʟ-galactonate to 2-keto-3-deoxygalactonate. SCO3486 showed maximum activity at pH 6.0 at 50℃, increased activity in the presence of iron ions, and activity against various aldehyde substrates, which is quite distinct from AHG-specific H2IFE7.1 in Vibrio sp. EJY3. Therefore, the catabolic pathway of AHG seems to be similar in most agar-degrading microorganisms, but the enzymes involved appear to be very diverse.
Keywords
Streptomyces coelicolor A3 (2); 3,6-anhydro-${\tiny\text{L}}$-galactose catabolism; 3,6-anhydro-${\tiny\text{L}}$-galactose dehydrogenase; 3,6-anhydro-${\tiny\text{L}}$-galactonate cycloisomerase; SCO3480; SCO3486;
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1 Temuujin U, Chi WJ, Chang YK, Hong SK. 2012. Identification and biochemical characterization of Sco3487 from Streptomyces coelicolor A3(2), an exo- and endo-type β-agarase-producing neoagarobiose. J. Bacteriol. 194: 142-149.   DOI
2 Green MR, Sambrook J. 2012. Molecular Cloning. A Laboratory Manual, 4th ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, USA.
3 Laemmli UK. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680-685.   DOI
4 Bradford MM.1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72: 248-254.   DOI
5 Asghar S, Lee CR, Chi WJ, Kang DK, Hong SK. 2019. Molecular cloning and characterization of a novel cold-adapted alkaline 1,3-α-3,6-anhydro-l-galactosidase, Ahg558, from Gayadomonas joobiniege G7. Appl. Biochem. Biotechnol. 188: 1077-1095.   DOI
6 Bentley SD, Chater KF, Cerdeno-Tarraga AM, Challis GL, Thomson NR, James KD, et al. 2002. Complete genome sequence of the model actinomycete Streptomyces coelicolor A3(2). Nature 417: 141-147.   DOI
7 Gerlt JA, Babbit, PC, Jacobson MP, Almo SC. 2012. Divergent evolution in the enolase superfamily: strategies for assigning functions. J. Biol. Chem. 287: 29-34.   DOI
8 Andberg M, Maaheimo H, Boer H, Penttila M, Koivula A, Richard P. 2012. Characterization of a novel Agrobacterium tumefaciens galactarolactone cycloisomerase enzyme for direct conversion of D-galactarolactone to 3-deoxy-2-keto-L-threo-hexarate. J. Biol. Chem. 287: 17662-17671.   DOI
9 Vasiliou V, Pappa A, Estay T. 2004. Role of human aldehyde dehydrogenases in endobiotic and xenobiotic metabolism. Drug Metab. Rev. 36: 279-299.   DOI
10 Chen Y, Mehta G, Vasiliou V. 2009. Antioxidant defenses in the ocular surface. Ocul. Surf. 7: 176-185.   DOI
11 Szumilo T. 1981. Pathway for D-galactonate catabolism in non-pathogenic mycobacteria. J. Bacteriol. 148: 368-370.   DOI
12 Wong TY, Yao XT. 1994. The DeLey-Doudoroff pathway of galactose metabolism in Azotobacter vinelandii. Appl. Environ. Microbiol. 60: 2065-2068.   DOI
13 Lee SB, Cho SJ, Kim JA, Lee SY, Kim SM, Lim HS. 2014. Metabolic pathway of 3,6-anhydro-L-galactose in agar-degrading microorganisms. Biotechnol. Bioproc. E. 19: 866-878.   DOI
14 Kieser T, Bibb MJ, Buttner MJ, Chater KF, Hopwood DA. 2000. Practical Streptomyces Genetics, John Innes Foundation, Norwich Research Park, Colney, Norwich NR4 7UH, England.
15 Petersen TN, Brunak S, von Heijne G, Nielsen H. 2011. SignalP 4.0: discriminating signal peptides from transmembrane regions. Nat. Methods 8: 785-786.   DOI
16 Chi WJ, Chang YK, Hong SK. 2012. Agar degradation by microorganisms and agar-degrading enzymes. Appl. Microbiol. Biotechnol. 94: 917-930.   DOI
17 Yun EJ, Yu S, Kim KH. 2017. Current knowledge on agarolytic enzymes and the industrial potential of agar-derived sugars. Appl. Microbiol. Biotechnol. 101: 5581-5589.   DOI
18 Hehemann JH, Correc G, Barbeyron T, Helbert W, Czjzek M, Michel G. 2010. Transfer of carbohydrate-active enzymes from marine bacteria to Japanese gut microbiota. Nature 464: 908-912.   DOI
19 Park SH, Lee CR, Hong SK. 2020. Implications of agar and agarase in industrial applications of sustainable marine biomass. Appl. Microbiol. Biotechnol. 104: 2815-2832.   DOI
20 Yun EJ, Lee S, Kim JH, Kim BB, Kim HT, Lee SH, et al. 2013. Enzymatic production of 3,6-anhydro-L-galactose from agarose and its purification and in vitro skin whitening and anti-inflammatory activities. Appl. Microbiol. Biotechnol. 97: 2961-2970.   DOI
21 Singh S, Brocker C, Jackson B, Matsumoto A, Thompson D. 2013. Aldehyde dehydrogenases in cellular response to oxidative/electrophilic stress. Free Radic. Biol. Med. 56: 89-101.   DOI
22 Yu S, Choi I-G, Yun EJ, Kim KH. 2018. High substrate specificity of 3,6-anhydro-l-galactose dehydrogenase indicates its essentiality in the agar catabolism of a marine bacterium, Process Biochem. 64: 130-135.   DOI
23 Wang Y, Li PY, Zhang Y, Cao HY, Wang YJ, Li CY, et al. 2020. 3,6-Anhydro-L-galactose dehydrogenase VvAHGD is a member of a new aldehyde dehydrogenase family and catalyzes by a novel mechanism with conformational switch of two catalytic residues Cysteine 282 and Glutamate 248. J. Mol. Biol. 432: 2186-2203.   DOI
24 Yun EJ, Lee AR, Kim JH, Cho KM, Kim KH. 2017. 3,6-Anhydro-L-galactose, a rare sugar from agar, a new anticariogenic sugar to replace xylitol. Food Chem. 221: 976-983.   DOI
25 Kim NJ, Li H, Jung K, Chang HN, Lee PC. 2011. Ethanol production from marine algal hydrolysates using Escherichia coli KO11. Bioresour. Technol. 107: 7466-7469.
26 Seo JW, Tsevelkhorloo M, Lee CR, Kim SH, Kang DK, Asghar S, et al. 2020. Molecular characterization of a novel 1,3-α-3,6-anhydro-l-galactosidase, ahg943, with cold- and high-salt-tolerance from Gayadomonas joobiniege G7. J. Microbiol. Biotechnol. 30: 1659-1669.   DOI
27 Stanier RY. 1942. Agar decomposing strains of the Actinomyces coelicolor species group. J. Bacteriol. 44: 555-570.   DOI
28 Temuujin U, Chi WJ, Lee SY, Chang YK, Hong SK. 2011. Overexpression and biochemical characterization of DagA from Streptomyces coelicolor A3(2): an endo-type β-agarase producing neoagarotetraose and neoagarohexaose. Appl. Microbiol. Biotechnol. 92: 749-759.   DOI
29 Yun EY, Lee HT, Kim KH. 2015. The novel catabolic pathway of 3,6-anhydro-L-galactose, the main component of red macroalgae, in a marine bacterium. Environ. Microbiol. 17: 1677-1688.   DOI
30 Lee S, Yun EJ, Kim KH, Kim HY, Choi IG. 2017. 3,6-Anhydro-L-galactonate cycloisomerase from Vibrio sp. strain EJY3: crystallization and X-ray crystallographic analysis. Acta Crystallogr. F Struct. Biol. Commun. 73: 511-514.   DOI