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The loop structure of Actinomycete glycoside hydrolase family 5 mannanases governs substrate recognition

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Title: The loop structure of Actinomycete glycoside hydrolase family 5 mannanases governs substrate recognition
Authors: Kumagai, Yuya Browse this author
Yamashita, Keitaro Browse this author
Tagami, Takayoshi Browse this author
Uraji, Misugi Browse this author
Wan, Kun Browse this author
Okuyama, Masayuki Browse this author →KAKEN DB
Yao, Min Browse this author →KAKEN DB
Kimura, Atsuo Browse this author →KAKEN DB
Hatanaka, Tadashi Browse this author
Keywords: Actinomycete
chimeric enzyme
glycoside hydrolase family 5
Issue Date: Oct-2015
Publisher: Wiley-Blackwell
Journal Title: Febs journal
Volume: 282
Issue: 20
Start Page: 4001
End Page: 4014
Publisher DOI: 10.1111/febs.13401
PMID: 26257335
Abstract: Endo--1,4-mannanases from Streptomyces thermolilacinus (StMan) and Thermobifida fusca (TfMan) demonstrated different substrate specificities. StMan hydrolyzed galactosylmannooligosaccharide (GGM5; 6(III),6(IV)--d-galactosyl mannopentaose) to GGM3 and M2, whereas TfMan hydrolyzed GGM5 to GGM4 and M1. To determine the region involved in the substrate specificity, we constructed chimeric enzymes of StMan and TfMan and evaluated their substrate specificities. Moreover, the crystal structure of the catalytic domain of StMan (StMandC) and the complex structure of the inactive mutant StE273AdC with M6 were solved at resolutions of 1.60 and 1.50 angstrom, respectively. Structural comparisons of StMandC and the catalytic domain of TfMan lead to the identification of a subsite around -1 in StMandC that could accommodate a galactose branch. These findings demonstrate that the two loops (loop7 and loop8) are responsible for substrate recognition in GH5 actinomycete mannanases. In particular, Trp281 in loop7 of StMan, which is located in a narrow and deep cleft, plays an important role in its affinity toward linear substrates. Asp310 in loop8 of StMan specifically bound to the galactosyl unit in the -1 subsite.
Type: article (author version)
Appears in Collections:農学院・農学研究院 (Graduate School of Agriculture / Faculty of Agriculture) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

Submitter: 熊谷 祐也

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