caa a22 4500 605506299 CHVBK 20210128100626.0 cr unu---uuuuu 210128e20150601xx s 000 0 eng 10.1007/s00253-014-6247-9 doi (NATIONALLICENCE)springer-10.1007/s00253-014-6247-9 Immobilization on macroporous resin makes E. coli RutB a robust catalyst for production of (−) Vince lactam [Elektronische Daten] [Jianjun Wang, Junge Zhu, Sheng Wu] A novel (+) γ-lactamase gene (rutB) was cloned from Escherichia coli JM109 and expressed in E. coli BL21 (DE3), and the recombinant protein was characterized. The optimal conditions for the enzyme were pH 7.0 and temperature 30°C, which indicated that it was a mesophilic protein. The free purified enzyme was deactivated when incubated at 50°C for 30min. However, the k cat value of RutB at its optimal temperature was about 2.5 times that of the archaeal enzyme from Sulfolobus sofataricus at its optimal temperature (85°C). After immobilization on macroporous resin using glutaraldehyde cross-linkage, the thermostability of the crude enzyme was greatly enhanced and the deactivating temperature was raised to 70°C. After immobilization, the minimal substrate inhibition concentration for RutB also improved from 0.75 to 1.5M. The optimal concentrations of immobilized enzyme and substrate were determined to be 250mg/ml and 1.5M, when the initial reaction velocity was the response variable in batch transformations. This immobilization of RutB on macroporous resins provides another feasible approach for the preparation of optically active Vince lactam. As a member of the isochorismatase superfamily, RutB was demonstrated to be another typical γ-lactamase that showed catalytic promiscuity. Springer-Verlag Berlin Heidelberg, 2014 (+) γ-lactamase nationallicence Glutaraldehyde cross-linkage nationallicence Immobilization nationallicence RutB nationallicence Wang Jianjun State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, No 1 Beichen West Road, 100101, Beijing, Chaoyang, People's Republic of China aut Zhu Junge State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, No 1 Beichen West Road, 100101, Beijing, Chaoyang, People's Republic of China aut Wu Sheng State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, No 1 Beichen West Road, 100101, Beijing, Chaoyang, People's Republic of China aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/11(2015-06-01), 4691-4700 0175-7598 99:11<4691 2015 99 253 https://doi.org/10.1007/s00253-014-6247-9 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-014-6247-9 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Wang Jianjun State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, No 1 Beichen West Road, 100101, Beijing, Chaoyang, People's Republic of China aut NATIONALLICENCE 700 1- Zhu Junge State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, No 1 Beichen West Road, 100101, Beijing, Chaoyang, People's Republic of China aut NATIONALLICENCE 700 1- Wu Sheng State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, No 1 Beichen West Road, 100101, Beijing, Chaoyang, People's Republic of China aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/11(2015-06-01), 4691-4700 0175-7598 99:11<4691 2015 99 253