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Identification of GH15 Family Thermophilic Archaeal Trehalases That Function within a Narrow Acidic-pH Range.
Appl Environ Microbiol 2015; 81(15):4920-31AE

Abstract

Two glucoamylase-like genes, TVN1315 and Ta0286, from the archaea Thermoplasma volcanium and T. acidophilum, respectively, were expressed in Escherichia coli. The gene products, TVN1315 and Ta0286, were identified as archaeal trehalases. These trehalases belong to the CAZy database family GH15, although they have putative (α/α)6 barrel catalytic domain structures similar to those of GH37 and GH65 family trehalases from other organisms. These newly identified trehalases function within a narrow range of acidic pH values (pH 3.2 to 4.0) and at high temperatures (50 to 60°C), and these enzymes display Km values for trehalose higher than those observed for typical trehalases. These enzymes were inhibited by validamycin A; however, the inhibition constants (Ki) were higher than those of other trehalases. Three TVN1315 mutants, corresponding to E408Q, E571Q, and E408Q/E571Q mutations, showed reduced activity, suggesting that these two glutamic acid residues are involved in trehalase catalysis in a manner similar to that of glucoamylase. To date, TVN1315 and Ta0286 are the first archaeal trehalases to be identified, and this is the first report of the heterologous expression of GH15 family trehalases. The identification of these trehalases could extend our understanding of the relationships between the structure and function of GH15 family enzymes as well as glycoside hydrolase family enzymes; additionally, these enzymes provide insight into archaeal trehalose metabolism.

Authors+Show Affiliations

Department of Chemistry and Life Science, Kogakuin University, Hachioji, Tokyo, Japan bt11532@ns.kogakuin.ac.jp.Department of Chemistry and Life Science, Kogakuin University, Hachioji, Tokyo, Japan.Department of Chemistry and Life Science, Kogakuin University, Hachioji, Tokyo, Japan.Department of Chemistry and Life Science, Kogakuin University, Hachioji, Tokyo, Japan.Department of Chemistry and Life Science, Kogakuin University, Hachioji, Tokyo, Japan.Department of Chemistry and Life Science, Kogakuin University, Hachioji, Tokyo, Japan.Department of Chemistry and Life Science, Kogakuin University, Hachioji, Tokyo, Japan.Department of Chemistry and Life Science, Kogakuin University, Hachioji, Tokyo, Japan Stem Cell Project, Tokyo Metropolitan Institute of Medical Science, Setagaya-ku, Tokyo, Japan.

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

25979886

Citation

Sakaguchi, Masayoshi, et al. "Identification of GH15 Family Thermophilic Archaeal Trehalases That Function Within a Narrow Acidic-pH Range." Applied and Environmental Microbiology, vol. 81, no. 15, 2015, pp. 4920-31.
Sakaguchi M, Shimodaira S, Ishida SN, et al. Identification of GH15 Family Thermophilic Archaeal Trehalases That Function within a Narrow Acidic-pH Range. Appl Environ Microbiol. 2015;81(15):4920-31.
Sakaguchi, M., Shimodaira, S., Ishida, S. N., Amemiya, M., Honda, S., Sugahara, Y., ... Kawakita, M. (2015). Identification of GH15 Family Thermophilic Archaeal Trehalases That Function within a Narrow Acidic-pH Range. Applied and Environmental Microbiology, 81(15), pp. 4920-31. doi:10.1128/AEM.00956-15.
Sakaguchi M, et al. Identification of GH15 Family Thermophilic Archaeal Trehalases That Function Within a Narrow Acidic-pH Range. Appl Environ Microbiol. 2015;81(15):4920-31. PubMed PMID: 25979886.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Identification of GH15 Family Thermophilic Archaeal Trehalases That Function within a Narrow Acidic-pH Range. AU - Sakaguchi,Masayoshi, AU - Shimodaira,Satoru, AU - Ishida,Shin-Nosuke, AU - Amemiya,Miko, AU - Honda,Shotaro, AU - Sugahara,Yasusato, AU - Oyama,Fumitaka, AU - Kawakita,Masao, Y1 - 2015/05/15/ PY - 2015/03/24/received PY - 2015/05/09/accepted PY - 2015/5/17/entrez PY - 2015/5/17/pubmed PY - 2016/3/30/medline SP - 4920 EP - 31 JF - Applied and environmental microbiology JO - Appl. Environ. Microbiol. VL - 81 IS - 15 N2 - Two glucoamylase-like genes, TVN1315 and Ta0286, from the archaea Thermoplasma volcanium and T. acidophilum, respectively, were expressed in Escherichia coli. The gene products, TVN1315 and Ta0286, were identified as archaeal trehalases. These trehalases belong to the CAZy database family GH15, although they have putative (α/α)6 barrel catalytic domain structures similar to those of GH37 and GH65 family trehalases from other organisms. These newly identified trehalases function within a narrow range of acidic pH values (pH 3.2 to 4.0) and at high temperatures (50 to 60°C), and these enzymes display Km values for trehalose higher than those observed for typical trehalases. These enzymes were inhibited by validamycin A; however, the inhibition constants (Ki) were higher than those of other trehalases. Three TVN1315 mutants, corresponding to E408Q, E571Q, and E408Q/E571Q mutations, showed reduced activity, suggesting that these two glutamic acid residues are involved in trehalase catalysis in a manner similar to that of glucoamylase. To date, TVN1315 and Ta0286 are the first archaeal trehalases to be identified, and this is the first report of the heterologous expression of GH15 family trehalases. The identification of these trehalases could extend our understanding of the relationships between the structure and function of GH15 family enzymes as well as glycoside hydrolase family enzymes; additionally, these enzymes provide insight into archaeal trehalose metabolism. SN - 1098-5336 UR - https://www.unboundmedicine.com/medline/citation/25979886/Identification_of_GH15_Family_Thermophilic_Archaeal_Trehalases_That_Function_within_a_Narrow_Acidic_pH_Range_ L2 - http://aem.asm.org/cgi/pmidlookup?view=long&pmid=25979886 DB - PRIME DP - Unbound Medicine ER -