|
|
|
| Isolation and Fermentation Optimization of a (+)γ-Lactamase-producing Strain |
| CHEN Hong-Gan, NI Ye, SUN Zhi-Hao |
| Key Lab of Industrial Biotechnology, Ministry of Education, Jiangnan University, School of Biotechnology, Wuxi 214122, China |
|
|
|
Abstract A (+)γ-lactamase-producing microorganisms with high enantioselectivity was isolated from soil, and was identified and deposited as Delftia sp.CGMCC No.5755. The optimized fermentation medium consists of: 30 g/L sucrose, 30 g/L peptone, 25 g/L beef extract, 5 g/L acetamide, and 1 g/L MgSO4; and the optimum temperature and initial pH are 32℃ and pH 7.0, respectively. Under the optimized conditions, 15.6 g/L of Delftia cells and 687 U/L of (+)γ-lactamase were obtained after 20 h of fermentation. Biotransformation using Delftia resting cells was carried out at 100 g/L of (±)γ-lactam substrate, the optical purify of product (-)γ-lactam reached over 99.9%e.e., and the conversion rate was 53.7% after 12h.The study provides a feasible approach for the biocatalytic preparation of (-)γ-lactam.
|
|
Received: 08 June 2012
Published: 25 September 2012
|
|
|
|
|
[1] Brabban A D,Littlechild J,Wisdom R. Stereospecific γ-lactamase activity in a Pseudomonas fluorescensspecies. Journal of industrial Microbiology,1996,16(1):8-14.
[2] Toogood H S,Brown R C,Line K,et al. The use of a thermostable signature amidase in the resolution of the bicyclic synthon (rac)γ-lactam. Tetrahedron,2004,60 (3):711-716.
[3] Taylor S J C,Sutherland A G,Lee C,et al. Chemoenzymatic synthesis of (-)-carbovir utilizing a whole cell catalysed resolution of 2-azabicyclo hept-5-en-3-one. Journal of the Chemical Society:Chemical Communications,1990,16(1):1120-1121.
[4] Chand P,Kotian P L,Dehghani A,et al. Systematic structure-based design and stereoselective synthesis of novel multisubstitutedcyclopentane derivatives with potent antiinfluenzaactivity. Journal of medicinal chemistry,2001,44(25):4379-4392.
[5] Lagoja I M,Clercq E D. Anti-influenzavirus agents:synthesis and mode of action. Medicinal Research Reviews,2008,28(1):1-38.
[6] 王建军,郑国钧,吴胜.微生物来源的γ-内酰胺酶的研究进展.微生物学报,2010,50(8):988-994. Wang J J, Zheng G J, Wu S. Advances in lactamase from microbes-A review. Acta Microbiologica Sinica,2010,50(8):988-994.
[7] Karen E H T. (+)-and (-)-2-Azabicyclo hept-5-en-3-one Extremely useful synthons. Chemsitry Today,2009,27(4):23-25.
[8] Chirotech Technology,Ltd.,United Kingdom. Microorganism,lactamase enzyme obtained therefrom,and their use:United States patent,US006090616A. 2000-7-18.
[9] Taylor S J C,Brown R C,Keene P A,et al. Novel Screening Methods-The Key to Cloning Commercially Successful Biocatalysts. Bioorganic and Medicinal Chemistry,1999,7(10):2163-2168.
[10] Hickey A M,Ngamsom B,Wiles C,et al. A microreactor for the study of biotransformations by a cross-linked γ-lactamase enzyme. Biotechnology journal,2009,4(4):510-516.
[11] 李海泉,苏磊,杨柳,等.产γ-内酰胺水解酶菌株的筛选及发酵条件研究.微生物学报,2006,46(4):571-575. Li H Q, Su L, Yang L, et al. Study on the screening of lactamase and its fermentation conditions. Acta Microbiologica Sinica,2006,46(4):571-575.
[12] Wang J J, Guo X Y, Zheng G J. Purification and characterization of an novel (-)gamma-lactamase from Microbacterium hydrocarbonoxydans. Annals of Microbiology,2009,59(2):345-348.
[13] Yang M, Gao Q,Wu S,et al. Characterization of a recombinant (-)γ-lactamase from Microbacterium hydrocarbonoxydans. Biotechnology Letters,2012,34(2):275-279.
[14] 北京化工大学.微杆菌及诱变方法与其在手性药物生产上的应用:中国,CN101240257A. 2008-8-30. Beijing University of Chemical Technology. Microbacterium,mutation method and its application in synthesis of chiral pharmaceutical:China, CN101240257A. 2008-8-30.
[15] Qin X X, Wang J J, Zheng G J. Enantioselective Resolution of γ-Lactam by a Whole Cell of Microbacterium hydrocarbonoxydans (L29-9)Immobilized in Polymer of PVA-Alginate-Boric Acid. Appl Biochem Biotechnol,2010,162(8):2345-2354.
[16] 中国科学院微生物研究所.一种具有拆分消旋体γ-内酰胺活性的(+)γ-内酰胺酶及编码基因与应用:中国,CN 1019217422010A. 2010-12-22. Institute of Microbiology,Chinese Academy of Sciences. A-Lactamase for enantioselective resolution of-Lactam,coding gene and its use:China, CN1019217422010A. 2010-12-22.
[17] Altshchul S F,Madden T L,Schaffer A A,et al. Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Research,1997,25(17):3389-3402. |
|
Viewed |
|
|
|
Full text
|
|
|
|
|
Abstract
|
|
|
|
|
Cited |
|
|
|
|
| |
Shared |
|
|
|
|
| |
Discussed |
|
|
|
|
