|
|
|
| The Mutasynthetic Production of CHCB1 from a Streptomyces avermitilis Mutant in the Presence of Cyclohexanecarboxylic Acid |
| JIANG Wei1,2, HONG Xiang2, ZHANG Xiao-Lin2, GUO Wei-Qun2, LIU Di1 |
1.College of Life Science,Northeast Agricultural University,Harbin150030,China
2.Academy of State Administration of Grain,Beijing100037,China |
|
|
|
Abstract Gene deletion vector pLJ04(pKC1139∷△bkdF+△bkdH)was used to disrupt bkdFGH in Streptomyces avermitilis 76-02-e, an industrial producer of anthelmintic avermectin. The disruptants were confirmed by PCR. Shake flask experiment and HPLC analysis showed that the mutant lost the ability to produce avermectins. As it is expected, the mutant, named S.avermitilis bkd76-3, could restore the ability of producing avermectins when the feeding of methylbutyric acid and isobutyric acid to its fermentations was carried out. The addition of cyclohexanecarboxylic acid (CHC) into fermentations of the S.avermitilis bkd76-3 allowed for production of four components, two of which was confirmed as CHC-B1 and CHC-A2 by LC/MS analysis, respectively.
|
|
Received: 27 March 2009
Published: 28 July 2009
|
|
|
|
|
| ［1］ 白宝星，张春燕，田敏，等.突变生物合成在微生物药物领域的研究进展.中国抗生素杂志，2007，32(2):65~73
Bai B X, Zhang CH Y, Tian M, et al.Chinese Journal of Antibiotics, 2007, 32(2):65~73
［2］ 房春林，杨光有，古小彬.多拉菌素的研究进展.中国畜牧兽医，2006，33(5):55~57
Fang C L, Yang Y G, Gu X B, et al.China Animal Husbandry & Veterinary Medicine, 2006, 33(5):55~57
［3］ Burg R W, Miller B M,Baker E E,et al.Avermectins,new family of potent anthelmintic agents:producing organism and fermentation.Antimicorb Agents Chemother,1979, 15 :361
［4］ Claudio D D, Ronald W F, Edmund W H, et al. A second branchedchain alphaketo acid dehydrogenase gene cluster (bkdFGH) from Streptomyces avermitilis: its relationship to avermectin biosynthesis and construction of a bkdF mutant suitable for production of novel antiparasitic avermectins. J. Bacteriol,1995, 177: 3504~3511
［5］ Cropp T A, Dennis J W, Reynolds K A. Identification of a cyclohexylcarbonyl CoA biosynthetic gene cluster and application in the production of doramectin. Nature, 2000, 18:980~983
［6］ Hanahan D. Studies on transformation of Escherichia coli with plasmids. J Mot Biol,1983,166:557
［7］ 郭伟群.阿维链霉菌高产菌株的选育和发酵条件的优化.北京:中国农业大学，2007
Guo W Q. Screening of high avermectinproducing strain and optimizes on the fermentation condition.Beijing: China Agricultural University, 2007
［8］ Bierman M, Logan R, O'Brien K. Plasmid cloning vectors for the conjugal transfer of DNA from Escherichia coli to Streptomyces spp. Gene, 1992, 116:43~49
［9］ Sambrook J, Fritsch E F, Maniatis T. Molecular Cloning. A Laboratory Manual.2nded. New York: Cold Spring Harbor laboratory Press, 1989
［10］ MacNeil D J, Klapko L M. Transformation of Streptomyces avermitilis by plasmid DNA. J Ind Microbiol, 1987,2: 209~218
［11］ Ikeda H, Kotaki H, Tanaka H, et al. Involvement of glucose catabolism in avermectin production by Streptomyces avermitilis. Antimicrob Agents Chemother, 1988,32:282~284
［12］ Hopwood D A, Bibb M J, Chater K F, et al. Genetic manipulation of Streptomyces. A Laboratory Manual. Norwich: John Innes Foundation, 1985
［13］ 宋渊，曹贵明，陈芝，等.阿维菌素高产菌株的选育及阿维菌素B1的鉴定.生物工程学报，2000，16(1):31~35
Song Y, Cao G M, Chen Z.Chinese Journal of Biotechnology,2000,16(1):31~35
［14］ Christopher J D, Stephen P G, Alexander C G, et al. Novel avermectins produced by mutational biosynthesis. The Journal of Antibiotics, 1991,44:357~365 |
|
Viewed |
|
|
|
Full text
|
|
|
|
|
Abstract
|
|
|
|
|
Cited |
|
|
|
|
| |
Shared |
|
|
|
|
| |
Discussed |
|
|
|
|
