title: Enzymatic characterization of two acetyl-CoA synthetase genes from Populus trichocarpa.
abstract: The acetyl-CoA synthetase (ACS) family is a subfamily of adenylate-forming enzymes, which has a close evolutionary relationship with the 4-coumarate:CoA ligase (4CL) family. In this study, two ACS genes were cloned from Populus trichocarpa and were named PtrACS1 and PtrACS2. Bioinformatics characterization of PtrACS1 and PtrACS2 showed that they contained the key ACS residues and a putative peroxisome targeting sequence 1 (PTS1) at the end of the C-terminal sequence. Real-time PCR results showed that PtrACS1 and PtrACS2 were expressed in the phloem, xylem, leaves, and roots of one-year-old P. trichocarpa, but were expressed primarily in the leaves. The ACS enzyme activity was higher in leaves than other tissues in P. trichocarpa. Two overexpressed recombinant proteins showed no catalytic activity toward the substrates of 4CL, but did have notable catalytic activity toward sodium acetate and substrates of ACS. The relative activities of PtrACS1 and PtrACS2 were 194.16 ± 11.23 and 422.25 ± 21.69 μM min(-1) mg(-1), respectively. The K m and V max of PtrACS1 were 0.25 mM and 698.85 μM min(-1) mg(-1), while those for PtrACS2 were 0.72 mM and 245.96 μM min(-1) mg(-1), respectively. Our results revealed that both proteins belong to the ACS family, and provide a theoretical foundation for the identification and functional analysis of members of the adenylate-forming enzyme superfamily.
authors.affiliation: College of Life Sciences and Biotechnology, Beijing Forestry University, No. 35 Qinghua East Road, Haidian District, Beijing, 100083 People's Republic of China.|National Engineering Laboratory for Tree Breeding, Beijing, 100083 People's Republic of China.
authors.forename: Shan
authors.initials: S
authors.lastname: Cao
authors.affiliation: College of Life Sciences and Biotechnology, Beijing Forestry University, No. 35 Qinghua East Road, Haidian District, Beijing, 100083 People's Republic of China.|National Engineering Laboratory for Tree Breeding, Beijing, 100083 People's Republic of China.
authors.forename: Hui
authors.initials: H
authors.lastname: Li
authors.affiliation: College of Life Sciences and Biotechnology, Beijing Forestry University, No. 35 Qinghua East Road, Haidian District, Beijing, 100083 People's Republic of China.|National Engineering Laboratory for Tree Breeding, Beijing, 100083 People's Republic of China.
authors.forename: Xiaoyun
authors.initials: X
authors.lastname: Yao
authors.affiliation: College of Life Sciences and Biotechnology, Beijing Forestry University, No. 35 Qinghua East Road, Haidian District, Beijing, 100083 People's Republic of China.|National Engineering Laboratory for Tree Breeding, Beijing, 100083 People's Republic of China.
authors.forename: Lihong
authors.initials: L
authors.lastname: Li
authors.affiliation: College of Life Sciences and Biotechnology, Beijing Forestry University, No. 35 Qinghua East Road, Haidian District, Beijing, 100083 People's Republic of China.|National Engineering Laboratory for Tree Breeding, Beijing, 100083 People's Republic of China.
authors.forename: Luyao
authors.initials: L
authors.lastname: Jiang
authors.affiliation: College of Life Sciences and Biotechnology, Beijing Forestry University, No. 35 Qinghua East Road, Haidian District, Beijing, 100083 People's Republic of China.|National Engineering Laboratory for Tree Breeding, Beijing, 100083 People's Republic of China.
authors.forename: Qiang
authors.initials: Q
authors.lastname: Zhang
authors.affiliation: College of Life Sciences and Biotechnology, Beijing Forestry University, No. 35 Qinghua East Road, Haidian District, Beijing, 100083 People's Republic of China.|National Engineering Laboratory for Tree Breeding, Beijing, 100083 People's Republic of China.
authors.forename: Jiaxue
authors.initials: J
authors.lastname: Zhang
authors.affiliation: College of Life Sciences and Biotechnology, Beijing Forestry University, No. 35 Qinghua East Road, Haidian District, Beijing, 100083 People's Republic of China.
authors.forename: Di
authors.initials: D
authors.lastname: Liu
authors.affiliation: College of Life Sciences and Biotechnology, Beijing Forestry University, No. 35 Qinghua East Road, Haidian District, Beijing, 100083 People's Republic of China.|National Engineering Laboratory for Tree Breeding, Beijing, 100083 People's Republic of China.
authors.forename: Hai
authors.initials: H
authors.lastname: Lu
keywords: Acetyl-CoA synthetase (ACS); Enzyme activity; Populus trichocarpa; Prokaryotic expression