Increasing production of spinosad in Saccharopolyspora spinosa by metabolic engineering

被引:4
作者
Bridget, Adzemye Fovennso [1 ]
Nguyen, Chung Thanh [1 ]
Magar, Rubin Thapa [1 ]
Sohng, Jae Kyung [1 ,2 ,3 ]
机构
[1] Sun Moon Univ, Inst Biomol Reconstruct iBR, Dept Life Sci & Biochem Engn, Asan, Chungnam, South Korea
[2] Sun Moon Univ, Dept BT Convergent Pharmaceut Engn, Asan, Chungnam, South Korea
[3] Sun Moon Univ, Inst Biomol Reconstruct iBR, Dept Life Sci & Biochem Engn, 70 Sun Moon Ro,221 Tangjeong Myeon, Asan 31460, Chungnam, South Korea
基金
新加坡国家研究基金会;
关键词
metabolic engineering; pIBR-SPN; Saccharopolyspora spinosa; spinosad; BIOSYNTHETIC GENE-CLUSTER; CONTROL AGENTS; IMPROVEMENT; ENHANCEMENT; EXPRESSION; METK1-SP; PATHWAY; PCR;
D O I
10.1002/bab.2418
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Spinosad, a combination of spinosyn A and D produced by Saccharopolyspora spinosa, is a highly efficient pesticide. There has been a considerable interest in the improvement of spinosad production because of a low yield achieved by wild-type S. spinosa. In this study, we designed and constructed a pIBR-SPN vector. pIBR-SPN is an integrative vector that can be used to introduce foreign genes into the chromosome of S. spinosa. Different combinations of genes encoding forasamine and rhamnose were synthesized and used for the construction of different recombinant plasmids. The following recombinant strains were developed: S. spinosa pIBR-SPN (only the vector), S. spinosa pIBR-SPN F (forosamine genes), S. spinosa pIBR-SPN R (rhamnose genes), S. spinosa pIBR-SPN FR (forosamine and rhamnose genes), S. spinosa pIBR-SPN FRS (forosamine, rhamnose, and SAM [S-adenosyl-L-methionine synthetase] genes), and S. spinosa MUV pIBR-SPN FR. Among these recombinant strains, S. spinosa pIBR-SPN FR produced 1394 +/- 163 mg/L spinosad, which was 13-fold higher than the wild-type. S. spinosa MUV pIBR-SPN FR produced 1897 (+/- 129) mg/L spinosad, which was seven-fold higher than S. spinosa MUV and 17-fold higher than the wild-type strain.
引用
收藏
页码:1035 / 1043
页数:9
相关论文
共 43 条
  • [1] An Insight into the "-Omics" Based Engineering of Streptomycetes for Secondary Metabolite Overproduction
    Chaudhary, Amit Kumar
    Dhakal, Dipesh
    Sohng, Jae Kyung
    [J]. BIOMED RESEARCH INTERNATIONAL, 2013, 2013
  • [2] Characterization of the chromosomal integration of Saccharopolyspora plasmid pCM32 and its application to improve production of spinosyn in Saccharopolyspora spinosa
    Chen, Jian
    Xia, Haiyang
    Dang, Fujun
    Xu, Qingyu
    Li, Wenjun
    Qin, Zhongjun
    [J]. APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 2015, 99 (23) : 10141 - 10149
  • [3] DeAmicis CV, 1997, ACS SYM SER, V658, P144
  • [4] Stimulation of polyketide metabolism in Streptomyces fradiae by tylosin and its glycosylated precursors
    Fish, SA
    Cundliffe, E
    [J]. MICROBIOLOGY-UK, 1997, 143 : 3871 - 3876
  • [5] PCR-targeted Streptomyces gene replacement identifies a protein domain needed for biosynthesis of the sesquiterpene soil odor geosmin
    Gust, B
    Challis, GL
    Fowler, K
    Kieser, T
    Chater, KF
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2003, 100 (04) : 1541 - 1546
  • [6] In vitro characterization of the enzymes involved in TDP-D-forosamine biosynthesis in the spinosyn pathway of Saccharopolyspora spinosa
    Hong, Lin
    Zhao, Zongbao
    Melancon, Charles E., III
    Zhang, Hua
    Liu, Hung-Wen
    [J]. JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2008, 130 (14) : 4954 - 4967
  • [7] Improvement of Spinosad Production upon Utilization of Oils and Manipulation of β-Oxidation in a High-Producing Saccharopolyspora spinosa Strain
    Huang, Ying
    Zhang, Xiaolin
    Zhao, Chen
    Zhuang, Xuhui
    Zhu, Lin
    Guo, Chao
    Song, Yuan
    [J]. JOURNAL OF MOLECULAR MICROBIOLOGY AND BIOTECHNOLOGY, 2018, 28 (02) : 53 - 64
  • [8] Metabolic Engineering of Rational Screened Saccharopolyspora spinosa for the Enhancement of Spinosyns A and D Production
    Jha, Amit Kumar
    Pokhrel, Anaya Raj
    Chaudhary, Amit Kumar
    Park, Seong-Whan
    Cho, Wan Je
    Sohng, Jae Kyung
    [J]. MOLECULES AND CELLS, 2014, 37 (10) : 727 - 733
  • [9] Enhancement of Herboxidiene Production in Streptomyces chromofuscus ATCC 49982
    Jha, Amit Kumar
    Lamichhane, Janardan
    Sohng, Jae Kyung
    [J]. JOURNAL OF MICROBIOLOGY AND BIOTECHNOLOGY, 2014, 24 (01) : 52 - 58
  • [10] Enhanced Production of Spinosad in Saccharopolyspora spinosa by Genome Shuffling
    Jin, Z. H.
    Xu, B.
    Lin, S. Z.
    Jin, Q. C.
    Cen, P. L.
    [J]. APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY, 2009, 159 (03) : 655 - 663