Journal of Biological Chemistry p. 1239 - 1248 (2010)
Update date:2022-08-11
Topics:
Yasuhira, Kengo
Shibata, Naoki
Mongami, Go
Uedo, Yuki
Atsumi, Yu
Kawashima, Yasuyuki
Hibino, Atsushi
Tanaka, Yusuke
Lee, Young-Ho
Kato, Dai-Ichiro
Takeo, Masahiro
Higuchi, Yoshiki
Negoro, Seiji
We performed x-ray crystallographic analyses of the 6-aminohexanoate cyclic dimer (Acd) hydrolase (NylA) from Arthrobacter sp., an enzyme responsible for the degradation of the nylon-6 industry byproduct. The fold adopted by the 472-amino acid polypeptide generated a compact mixed α/β fold, typically found in the amidase signature superfamily; this fold was especially similar to the fold of glutamyl-tRNAGln amido-transferase subunit A (z score, 49.4) and malonamidase E2 (z score, 44.8). Irrespective of the high degree of structural similarity to the typical amidase signature superfamily enzymes, the specific activity of NylA for glutamine, malonamide, and indoleacetamide was found to be lower than 0.5% of that for Acd. However, NylA possessed carboxylesterase activity nearly equivalent to the Acd hydrolytic activity. Structural analysis of the inactive complex between the activity-deficient S174A mutant of NylA and Acd, performed at 1.8 A resolution, suggested the following enzyme/substrate interactions: a Ser174-cis-Ser150-Lys72 triad constitutes the catalytic center; the backbone N in Ala171 and Ala172 are involved in oxyanion stabilization; Cys316-Sγ forms a hydrogen bond with nitrogen (Acd-N7) at the uncleaved amide bond in two equivalent amide bonds of Acd. A single S174A, S150A, or K72A substitution in NylA by site-directed mutagenesis decreased the Acd hydrolytic and esterolytic activities to undetectable levels, indicating that Ser174-cis-Ser150-Lys72 is essential for catalysis. In contrast, substitutions at position 316 specifically affected Acd hydrolytic activity, suggesting that Cys316 is responsible for Acd binding. On the basis of the structure and functional analysis, we discussed the catalytic mechanisms and evolution of NylA in comparison with other Ser-reactive hydrolases.
View MoreYantai Derun Liquid Crystal Materials Co. Ltd.
website:http://www.ytderun.com
Contact:86-535-6300169
Address:ROOM 90, XIANGFU STREET, FUSHAN NEW-HIGH-TECH IDUSTRY ZONE, YANTAI
Shanghai Bojing Chemical Co., Ltd.
Contact:021-37122233
Address:6F Buildiing 11,No.388,Baifu Road,District Fengxian.Shanghai, China.
Hunan Zhongqi Pharmaceutical Co., Ltd
website:http://www.hnzqzy.com
Contact:0730-8722288 13807308622
Address:Wanjiafan Road ,Yueyang Economic And Technological Development Zone ,Hunan,PRC
Liaoning Yufeng Chemical Co.,Ltd.
Contact:86-0419-3418888
Address:The metallurgical industrial zone,shoushan town, Liaoyang, Liaoning, China
Xiamen Kaijia Imp & Exp Co., Ltd.
Contact:86-592-5101177
Address:Room406 Luhui Building No. 65 Haitian Road Huli Xiamen,China.
Doi:10.1016/0022-328X(94)80176-2
(1994)Doi:10.1016/j.molstruc.2015.01.050
(2015)Doi:10.1021/acs.macromol.9b02274
(2020)Doi:10.1039/c9cc00797k
(2019)Doi:10.1016/j.saa.2004.04.028
(2005)Doi:10.1021/j150498a017
(1952)