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Two trehalose-hydrolyzing enzymes from Crenarchaeon Sulfolobus acidocaldarius exhibit distinct activities and affinities toward trehalose.
Appl Microbiol Biotechnol 2018; 102(10):4445-4455AM

Abstract

Two archaeal trehalase-like genes, Saci1250 and Saci1816, belonging to glycoside hydrolase family 15 (GH15) from the acidophilic Crenarchaeon Sulfolobus acidocaldarius were expressed in Escherichia coli. The gene products showed trehalose-hydrolyzing activities, and the names SaTreH1 and SaTreH2 were assigned to Saci1816 and Saci1250 gene products, respectively. These newly identified enzymes functioned within a narrow range of acidic pH values at elevated temperatures, which is similar to the behavior of Euryarchaeota Thermoplasma trehalases. SaTreH1 displayed high KM and kcat values, whereas SaTreH2 had lower KM and kcat values despite a high degree of identity in their primary structures. A mutation analysis indicated that two glutamic acid residues in SaTreH1, E374 and E574, may be involved in trehalase catalysis because SaTreH1 E374Q and E574Q showed greatly reduced trehalose-hydrolyzing activities. Additional mutations substituting G573 and H575 residues with serine and glutamic acid residues, respectively, to mimic the TVN1315 sequence resulted in a decrease in trehalase activity and thermal stability. Taken together, the results indicated that Crenarchaea trehalases adopt active site structures that are similar to Euryarchaeota enzymes but have distinct molecular features. The identification of these trehalases could extend our understanding of the relationships between the structure and function of GH15 trehalases as well as other family enzymes and will provide insights into archaeal trehalose metabolism.

Authors+Show Affiliations

Department of Chemistry and Life Science, Kogakuin University, 2,665-1 Nakano-cho, Hachioji, Tokyo, 192-0015, Japan.Department of Chemistry and Life Science, Kogakuin University, 2,665-1 Nakano-cho, Hachioji, Tokyo, 192-0015, Japan.Department of Chemistry and Life Science, Kogakuin University, 2,665-1 Nakano-cho, Hachioji, Tokyo, 192-0015, Japan.Department of Chemistry and Life Science, Kogakuin University, 2,665-1 Nakano-cho, Hachioji, Tokyo, 192-0015, Japan.Department of Chemistry and Life Science, Kogakuin University, 2,665-1 Nakano-cho, Hachioji, Tokyo, 192-0015, Japan.Department of Chemistry and Life Science, Kogakuin University, 2,665-1 Nakano-cho, Hachioji, Tokyo, 192-0015, Japan. Stem Cell Project, Tokyo Metropolitan Institute of Medical Science, 2-1-6 Kami-kitazawa, Setagaya-ku, Tokyo, 156-8506, Japan.Department of Chemistry and Life Science, Kogakuin University, 2,665-1 Nakano-cho, Hachioji, Tokyo, 192-0015, Japan.Department of Chemistry and Life Science, Kogakuin University, 2,665-1 Nakano-cho, Hachioji, Tokyo, 192-0015, Japan. bt11532@ns.kogakuin.ac.jp.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

29574614

Citation

Yuasa, Mitsuhiro, et al. "Two Trehalose-hydrolyzing Enzymes From Crenarchaeon Sulfolobus Acidocaldarius Exhibit Distinct Activities and Affinities Toward Trehalose." Applied Microbiology and Biotechnology, vol. 102, no. 10, 2018, pp. 4445-4455.
Yuasa M, Okamura T, Kimura M, et al. Two trehalose-hydrolyzing enzymes from Crenarchaeon Sulfolobus acidocaldarius exhibit distinct activities and affinities toward trehalose. Appl Microbiol Biotechnol. 2018;102(10):4445-4455.
Yuasa, M., Okamura, T., Kimura, M., Honda, S., Shin, Y., Kawakita, M., ... Sakaguchi, M. (2018). Two trehalose-hydrolyzing enzymes from Crenarchaeon Sulfolobus acidocaldarius exhibit distinct activities and affinities toward trehalose. Applied Microbiology and Biotechnology, 102(10), pp. 4445-4455. doi:10.1007/s00253-018-8915-7.
Yuasa M, et al. Two Trehalose-hydrolyzing Enzymes From Crenarchaeon Sulfolobus Acidocaldarius Exhibit Distinct Activities and Affinities Toward Trehalose. Appl Microbiol Biotechnol. 2018;102(10):4445-4455. PubMed PMID: 29574614.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Two trehalose-hydrolyzing enzymes from Crenarchaeon Sulfolobus acidocaldarius exhibit distinct activities and affinities toward trehalose. AU - Yuasa,Mitsuhiro, AU - Okamura,Takeshi, AU - Kimura,Masahiro, AU - Honda,Shotaro, AU - Shin,Yongchol, AU - Kawakita,Masao, AU - Oyama,Fumitaka, AU - Sakaguchi,Masayoshi, Y1 - 2018/03/24/ PY - 2017/11/02/received PY - 2018/03/03/accepted PY - 2018/03/01/revised PY - 2018/3/27/pubmed PY - 2019/1/15/medline PY - 2018/3/26/entrez KW - Archaeal GH15 trehalase KW - Characterization KW - Crenarchaea KW - Expression KW - Sulfolobus acidocaldarius SP - 4445 EP - 4455 JF - Applied microbiology and biotechnology JO - Appl. Microbiol. Biotechnol. VL - 102 IS - 10 N2 - Two archaeal trehalase-like genes, Saci1250 and Saci1816, belonging to glycoside hydrolase family 15 (GH15) from the acidophilic Crenarchaeon Sulfolobus acidocaldarius were expressed in Escherichia coli. The gene products showed trehalose-hydrolyzing activities, and the names SaTreH1 and SaTreH2 were assigned to Saci1816 and Saci1250 gene products, respectively. These newly identified enzymes functioned within a narrow range of acidic pH values at elevated temperatures, which is similar to the behavior of Euryarchaeota Thermoplasma trehalases. SaTreH1 displayed high KM and kcat values, whereas SaTreH2 had lower KM and kcat values despite a high degree of identity in their primary structures. A mutation analysis indicated that two glutamic acid residues in SaTreH1, E374 and E574, may be involved in trehalase catalysis because SaTreH1 E374Q and E574Q showed greatly reduced trehalose-hydrolyzing activities. Additional mutations substituting G573 and H575 residues with serine and glutamic acid residues, respectively, to mimic the TVN1315 sequence resulted in a decrease in trehalase activity and thermal stability. Taken together, the results indicated that Crenarchaea trehalases adopt active site structures that are similar to Euryarchaeota enzymes but have distinct molecular features. The identification of these trehalases could extend our understanding of the relationships between the structure and function of GH15 trehalases as well as other family enzymes and will provide insights into archaeal trehalose metabolism. SN - 1432-0614 UR - https://www.unboundmedicine.com/medline/citation/29574614/Two_trehalose_hydrolyzing_enzymes_from_Crenarchaeon_Sulfolobus_acidocaldarius_exhibit_distinct_activities_and_affinities_toward_trehalose_ L2 - https://dx.doi.org/10.1007/s00253-018-8915-7 DB - PRIME DP - Unbound Medicine ER -