caa a22 4500 445818263 CHVBK 20180317145235.0 cr unu---uuuuu 170323e20110101xx s 000 0 eng 10.1007/s00792-010-0332-5 doi (NATIONALLICENCE)springer-10.1007/s00792-010-0332-5 Expression and characterization of a novel mesophilic protease from metagenomic library derived from Antarctic coastal sediment [Elektronische Daten] [Yingjing Zhang, Jing Zhao, Runying Zeng] A metagenomic cosmid library was constructed, in which the insert DNA was derived from the coastal sediment near Antarctic China Zhongshan Station. One clone (ACPRO001) expressing protease activity was isolated from the library using milk agar plates. Sequencing of the clone revealed a novel protease gene. The amino acid sequence comparison and phylogenetic analysis indicated that it could be classified as a subtilisin-like serine protease, though the highly conserved residue Asp was replaced by Ala. The ACPRO001 protease gene was expressed in pET-His and purified for characterization. The optimal temperature and pH for the activity of the ACPRO001 protease were 60°C and pH 9.0, respectively. The enzyme retained about 73% of residual activity after 2h incubation at 50°C in the presence of Ca2+. The presence of Ca2+ increased the thermostability of ACPRO001 protease obviously. The enzymatic activity was inhibited by 1mM phenylmethyl sulfonylfluoride (PMSF) and hydrochloride 4-(2-aminoethyl)-benzenesulfonyl fluoride (AEBSF), indicating that it was a serine protease. Springer, 2010 Protease nationallicence Metagenomic library nationallicence Antarctic nationallicence Zhang Yingjing Key Lab of Marine Biogenetic Resources, Third Institute of Oceanography, SOA, Daxue Road 178#, 361005, Xiamen, China aut Zhao Jing College of Oceanography and Environmental Science, Xiamen University, 361005, Xiamen, China aut Zeng Runying Key Lab of Marine Biogenetic Resources, Third Institute of Oceanography, SOA, Daxue Road 178#, 361005, Xiamen, China aut Extremophiles Springer Japan 15/1(2011-01-01), 23-29 1431-0651 15:1<23 2011 15 792 https://doi.org/10.1007/s00792-010-0332-5 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s00792-010-0332-5 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Zhang Yingjing Key Lab of Marine Biogenetic Resources, Third Institute of Oceanography, SOA, Daxue Road 178#, 361005, Xiamen, China aut NATIONALLICENCE 700 1- Zhao Jing College of Oceanography and Environmental Science, Xiamen University, 361005, Xiamen, China aut NATIONALLICENCE 700 1- Zeng Runying Key Lab of Marine Biogenetic Resources, Third Institute of Oceanography, SOA, Daxue Road 178#, 361005, Xiamen, China aut NATIONALLICENCE 773 0- Extremophiles Springer Japan 15/1(2011-01-01), 23-29 1431-0651 15:1<23 2011 15 792 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer