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PspAG97A: A Halophilic α-Glucoside Hydrolase with Wide Substrate Specificity from Glycoside Hydrolase Family 97

  • Li, Wei (School of Life Sciences, Anhui University) ;
  • Fan, Han (School of Life Sciences, Anhui University) ;
  • He, Chao (School of Life Sciences, Anhui University) ;
  • Zhang, Xuecheng (School of Life Sciences, Anhui University) ;
  • Wang, Xiaotang (Department of Chemistry and Biochemistry, Florida International University) ;
  • Yuan, Jing (School of Life Sciences, Anhui University) ;
  • Fang, Zemin (School of Life Sciences, Anhui University) ;
  • Fang, Wei (School of Life Sciences, Anhui University) ;
  • Xiao, Yazhong (School of Life Sciences, Anhui University)
  • Received : 2016.06.21
  • Accepted : 2016.08.02
  • Published : 2016.11.28

Abstract

A novel ${\alpha}-glucoside$ hydrolase (named PspAG97A) from glycoside hydrolase family 97 (GH97) was cloned from the deep-sea bacterium Pseudoalteromonas sp. K8, which was screened from the sediment of Kongsfjorden. Sequence analysis showed that PspAG97A belonged to GH97, and shared 41% sequence identity with the characterized ${\alpha}-glucoside$ BtGH97a. PspAG97A possessed three key catalytically related glutamate residues. Mutation of the glutamate residues indicated that PspAG97A belonged to the inverting subfamily of GH97. PspAG97A showed significant reversibility against changes in salt concentration. It exhibited halophilic ability and improved thermostability in NaCl solution, with maximal activity at 1.0 M NaCl/KCl, and retained more than 80% activity at NaCl concentrations ranging from 0.8 to 2.0 M for over 50 h. Furthermore, PspAG97A hydrolyzed not only ${\alpha}-1,4-glucosidic$ linkage, but also ${\alpha}-1,6-$ and ${\alpha}-1,2-glucosidic$ linkages. Interestingly, PspAG97A possessed high catalytic efficiency for long-chain substrates with ${\alpha}-1,6-linkage$. These characteristics are clearly different from other known ${\alpha}-glucoside$ hydrolases in GH97, implying that PspAG97A is a unique ${\alpha}-glucoside$ hydrolase of GH97.

Keywords

References

  1. Arnosti C. 2011. Microbial extracellular enzymes and the marine carbon cycle. Annu. Rev. Mar. Sci. 3: 401-425. https://doi.org/10.1146/annurev-marine-120709-142731
  2. Boutaiba S, Bhatnagar T, Hacene H, Mitchell D, Baratti J. 2006. Preliminary characterisation of a lipolytic activity from an extremely halophilic archaeon, Natronococcus sp. J. Mol. Catal. B Enzym. 41: 21-26. https://doi.org/10.1016/j.molcatb.2006.03.010
  3. Henrissat B, Davies G. 1997. Structural and sequence-based classification of glycoside hydrolases. Curr. Opin. Struct. Biol. 7: 637-644. https://doi.org/10.1016/S0959-440X(97)80072-3
  4. Hough DW, Danson MJ. 1999. Extremozymes. Curr. Opin. Chem. Biol. 3: 39-46. https://doi.org/10.1016/S1367-5931(99)80008-8
  5. Hughes C, Malki G, Loo C, Tanner A, Ganeshkumar N. 2003. Cloning and expression of ${\alpha}$-D-glucosidase and Nacetyl- ${\beta}$-glucosaminidase from the periodontal pathogen, Tannerella forsythensis (Bacteroides forsythus). Oral Microbiol. Immunol. 18: 309-312. https://doi.org/10.1034/j.1399-302X.2003.00091.x
  6. Kitamura M, Okuyama M, Tanzawa F, Mori H, Kitago Y, Watanabe N, et al. 2008. Structural and functional analysis of a glycoside hydrolase family 97 enzyme from Bacteroides thetaiotaomicron. J. Biol. Chem. 283: 36328-36337. https://doi.org/10.1074/jbc.M806115200
  7. Lombard V, Golaconda Ramulu H, Drula E, Coutinho PM, Henrissat B. 2014. The carbohydrate-active enzymes database (CAZy) in 2013. Nucleic Acids Res. 42: D490-D495. https://doi.org/10.1093/nar/gkt1178
  8. Médigue C, Krin E, Pascal G, Barbe V, Bernsel A, Bertin PN, et al. 2005. Coping with cold: the genome of the versatile marine Antarctica bacterium Pseudoalteromonas haloplanktis TAC125. Genome Res. 15: 1325-1335. https://doi.org/10.1101/gr.4126905
  9. Mevarech M, Frolow F, Gloss LM. 2000. Halophilic enzymes: proteins with a grain of salt. Biophys. Chem. 86: 155-164. https://doi.org/10.1016/S0301-4622(00)00126-5
  10. Naumoff DG. 2005. GH97 is a new family of glycoside hydrolases, which is related to the alpha-galactosidase superfamily. BMC Genomics 6: 112. https://doi.org/10.1186/1471-2164-6-112
  11. Noguchi A, Nakayama T, Hemmi H, Nishino T. 2003. Altering the substrate chain-length specificity of an ${\alpha}$-glucosidase. Biochem. Biophys. Res. Commun. 304: 684-690. https://doi.org/10.1016/S0006-291X(03)00647-8
  12. Oh E-J, Lee Y-J, Chol J, Seo MS, Lee MS, Kim GA, Kwon ST. 2008. Mutational analysis of Thermus caldophilus GK24 beta-glycosidase: role of His119 in substrate binding and enzyme activity. J. Microbiol. Biotechnol. 18: 287-294.
  13. Okuyama M. 2011. Function and structure studies of GH family 31 and 97 ${\alpha}$-glycosidases. Biosci. Biotechnol. Biochem. 75: 2269-2277. https://doi.org/10.1271/bbb.110610
  14. Okuyama M, Kitamura M, Hondoh H, Kang M-S, Mori H, Kimura A, et al. 2009. Catalytic mechanism of retaining ${\alpha}$-galactosidase belonging to glycoside hydrolase family 97. J. Mol. Biol. 392: 1232-1241. https://doi.org/10.1016/j.jmb.2009.07.068
  15. Okuyama M, Yoshida T, Hondoh H, Mori H, Yao M, Kimura A. 2014. Catalytic role of the calcium ion in GH97 inverting glycoside hydrolase. FEBS Lett. 588: 3213-3217. https://doi.org/10.1016/j.febslet.2014.07.002
  16. Pavlovic M, Dimitrijevic A, Bezbradica D, Milosavic N, Gavrovic-Jankulovic M, Segan D, Velickovic D. 2014. Dual effect of benzyl alcohol on ${\alpha}$-glucosidase activity: efficient substrate for high yield transglucosylation and noncompetitive inhibitor of its hydrolytic activity. Carbohydr. Res. 387: 14-18. https://doi.org/10.1016/j.carres.2013.08.028
  17. Rao L, Zhao X, Pan F, Li Y, Xue Y, Ma Y, Lu JR. 2009. Solution behavior and activity of a halophilic esterase under high salt concentration. PLoS One 4: e6980. https://doi.org/10.1371/journal.pone.0006980
  18. Saburi W, Rachi-Otsuka H, Hondoh H, Okuyama M, Mori H, Kimura A. 2015. Structural elements responsible for the glucosidic linkage-selectivity of a glycoside hydrolase family 13 exo-glucosidase. FEBS Lett. 589: 865-869. https://doi.org/10.1016/j.febslet.2015.02.023
  19. Shirai T, Hung VS, Morinaka K, Kobayashi T, Ito S. 2008. Crystal structure of GH13 ${\alpha}$-glucosidase GSJ from one of the deepest sea bacteria. Proteins 73: 126-133. https://doi.org/10.1002/prot.22044
  20. Tagami T, Yamashita K, Okuyama M, Mori H, Yao M, Kimura A. 2015. Structural advantage of sugar beet ${\alpha}$-glucosidase to stabilize the Michaelis complex with longchain substrate. J. Biol. Chem. 290: 1796-1803. https://doi.org/10.1074/jbc.M114.606939
  21. Zhang C, Kim S-K. 2010. Research and application of marine microbial enzymes: status and prospects. Marine Drugs 8: 1920-1934. https://doi.org/10.3390/md8061920

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