title: Sequence-based screening and characterization of cytosolic mandelate oxidase using oxygen as electron acceptor.
abstract: Sequence-based screening was carried out to find a type of cytosolic mandelate oxidase that converted l-mandelate to phenylglyoxylate using oxygen as the final electron acceptor. The sequence features of the cytosolic mandelate oxidase were summarized, and were used in the screening process. Mandelate oxidases from Streptomyces coelicolor (HmoSC) and Amycolatopsis orientalis (HmoAO) were screened and then they were heterologously expressed and characterized. At pH 7.3 40 °C, the HmoAO showed kcat and Km values of 140 min(-1) and 10.2 mM, the HmoSC showed kcat and Km values of 105.1 min(-1) and 2.06 mM. The HmoSC was thermal stable and retained its 90% activity at 60 °C for up to 5 h, while HmoAO lost most of its activity at this temperature. The HmoSC could effectively catalyze the conversion of l-mandelate to phenylglyoxylate at higher temperature using oxygen as the final electron acceptor.
authors: National Engineering Laboratory for Cereal Fermentation Technology, The Key Laboratory of Industrial Biotechnology of Ministry of Education, Jiangnan University, Wuxi 214122, PR China.
authors: Shuang Ping
authors: SP
authors: Liu
authors: National Engineering Laboratory for Cereal Fermentation Technology, The Key Laboratory of Industrial Biotechnology of Ministry of Education, Jiangnan University, Wuxi 214122, PR China.
authors: Rui Xia
authors: RX
authors: Liu
authors: National Engineering Laboratory for Cereal Fermentation Technology, The Key Laboratory of Industrial Biotechnology of Ministry of Education, Jiangnan University, Wuxi 214122, PR China.
authors: Liang
authors: L
authors: Zhang
authors: National Engineering Laboratory for Cereal Fermentation Technology, The Key Laboratory of Industrial Biotechnology of Ministry of Education, Jiangnan University, Wuxi 214122, PR China. Electronic address: gyshi@jiangnan.edu.cn.
authors: Gui Yang
authors: GY
authors: Shi
mesh_terms: D039903:Actinobacteria / Q000201:enzymology / Q000235:genetics; D000429:Alcohol Oxidoreductases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D003600:Cytosol / Q000201:enzymology; D004583:Electrons; D004795:Enzyme Stability; D006038:Glyoxylates / Q000378:metabolism; D007700:Kinetics; D008333:Mandelic Acids / Q000378:metabolism*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010100:Oxygen / Q000378:metabolism; D011487:Protein Conformation; D017386:Sequence Homology, Amino Acid; D048372:Streptomyces coelicolor / Q000201:enzymology / Q000235:genetics; D013696:Temperature
chemical_list: D001426:Bacterial Proteins; D006038:Glyoxylates; D008333:Mandelic Acids; C012482:phenylglyoxylic acid; D000429:Alcohol Oxidoreductases; C037938:mandelic acid; D010100:Oxygen
keywords: Amycolatopsis orientalis; Mandelate oxidase; Phenylglyoxylate; Streptomyces coelicolor