tailieunhanh - Expression of glutaryl7 aminocephalosporanic acid acylase in escherichia coli BL21(de3) and immobilization of recombinant enzyme on nanoporous materials

The synthesis of 7-ACA from cephalosporin C (CPC) by a two-step bioconversion using D-amino acid oxidase (DAAO) and glutaryl 7-ACA acylase (GLA) has been effectively and largely applied in pharmaceutical industry. In this study, the gene gla coding for 720-amino acid GLA from plasmid pUC57::gla was analyzed and successfully inserted into vector pET22b(+) to form expression vector pET22b(+)::gla. | Journal of Science and Technology 54 (4A) (2016) 123-131 EXPRESSION OF GLUTARYL7-AMINOCEPHALOSPORANIC ACID ACYLASE IN ESCHERICHIA COLI BL21(DE3) AND IMMOBILIZATION OF RECOMBINANT ENZYME ON NANOPOROUS MATERIALS Vu Thi Hanh Nguyen, Pham Thanh Huyen, Le Gia Hy, Phi Quyet Tien* Institute of Biotechnology, VAST, 18 Hoang Quoc Viet, Cau Giay, Hanoi, Vietnam * Email: tienpq@ Received: 15 August 2016; Accepted for publication: 5 October 2016 ABSTRACT The synthesis of 7-ACA from cephalosporin C (CPC) by a two-step bioconversion using D-amino acid oxidase (DAAO) and glutaryl 7-ACA acylase (GLA) has been effectively and largely applied in pharmaceutical industry. In this study, the gene gla coding for 720-amino acid GLA from plasmid pUC57::gla was analyzed and successfully inserted into vector pET22b(+) to form expression vector pET22b(+)::gla. The newly constructed expression vector pET22b(+)::gla was cloned and then transformed into Escherichia coli BL21(DE3) to generate recombinant strain E. coli BL21(DE3)[pET22b(+)::gla]. The suitable conditions for expression of gla gene were in LB medium at 30 oC and induced by mM of Isopropyl β-D-1thiogalactopyranoside (IPTG) for 3 hours. Under the chosen culturing parameters, expression of gla gene by E. coli BL21(DE3)/[pET22b(+)::gla] resulted in a recombinant GLA (rGLA) with molecular weight of 83 kDa and catalytic activity of U/mg of total protein. Experimental research on immobilization of rGLA onto ten nanoporous materials were showed that, SBA-15 was the best one for immobilization of rGLA, reaching activity of immobilized enzyme of U/g matrix. Furthermore, optimal conditions of procedure for immobilizing rGLA on nanomaterials (SBA-15) were determined as follows: temperature is 25 C, and immobilization time –60 minutes. Therefore the results reported in this study revealed the successfully heterologous expression of GLA in recombinant E. coli and potential immobilization of enzyme on inorganic .

• Hoá học
• Journal of Science and Technology
• Expression of glutaryl7 aminocephalosporanic acid acylase
• In escherichia coli BL21(de3)
• Immobilization of recombinant enzyme on nanoporous materials
• 7 ACA from cephalosporin C
• Journal of Marine science and Technology
• Marine science and Technology
• Marine science and Technology no 56
• Marine science and Technology year 2018
• Evaluation method
• Electrical energy management model
• Marine science and Technology no 53
• Diesel engines
• Journal of Fisheries science and Technology
• Fisheries science and Technology
• Fisheries science and Technology no 3
• Fisheries science and Technology year 2016
• Striped catfish
• Common carp cyprinus carpio
• Fisheries science and Technology no 4
• Fisheries science and Technology year 2018
• Coastal fi shing activities
• White striped cleaner shrimp
• Journal of Science
• Journal of Technology
• Technology Management
• Science and Technology
• International integration
• Science and Technology Strategy
• Evaluation of science and technology
• Global journal of computer science
• Global journal of computer science and technology
• Information & technology
• GIS based fire emergency response system
• Marine science and technology no 61
• Actual GPS receivers
• Display information on stability
• Nuclear science and technology
• The journal of Nuclear science and technology
• Rotary rack served for elemental concentration
• Seawaterusing co precipitation method
• Natural radionuclides
• Experimental data
• Low energy experiments
• Multigroup cross sections
• Stable isotopic values
• Radial blanket
• Dissolved nitrogen compounds
• Irradiation channels
• Neutron rich nuclei
• Prescila neutron probe
• Full core conversion
• Multipurpose research reactor
• Aquatic environment
• Geochemical parameters
• External reactor vessel cooling method
• Radiation crosslinked scaffolds
• Adipose derived stem cell culture
• Treatment plans
• Minor actinide compositions
• IM NAA of non standard geometrical samples
• Cross shaped obstacle
• University entrepreneurship
• Entrepreneurial research university
• Technology innovation
• Industry university cooperation
• Regional innovation
• Journal of technology and science education
• Bring science and technology closer to secondary school
• Technology closer to secondary school
• Secondary school teachers
• Face to face
• Technology development
• Technology commercialization
• The role of informal science centers in science education
• Informal science center
• Attudes toward science
• Scientfic thinking skills
• Scientific research
• Research quality
• Strategy and policy
• Research Organizations
• Policy Planning
• Science and Technology development
• Theory Z
• Human Resources
• Development policy of Japan
• ‘Standards’ on the bench
• Technological literacy render an adequate image of technology
• Adequate image of technology
• Standards for technological literacy
• Philosophy of technology
• The nature of technology
• Concepts of technology
• Post secondary project based learning in science
• Post secondary project based learning in technology
• Post secondary project based learning in engineering and mathematics
• Post secondary project