基因组重排选育Epothilone B高产菌株

生物技术通报 ›› 2013, Vol. 0 ›› Issue (5): 155-160.

基因组重排选育Epothilone B高产菌株

王大红¹, 刘飞²

(1.河南科技大学食品与生物工程学院，洛阳 471023；2.南阳师范学院生命科学与技术学院，南阳 473061)

收稿日期:2012-11-05 修回日期:2013-05-24 出版日期:2013-05-24 发布日期:2013-05-24
作者简介:王大红，男，博士，讲师，研究方向：发酵工程和微生物制药；E-mail：wangdahong2003@163.com
基金资助:
河南科技大学青年科学基金项目(2012QN003)

Breeding of High Epothilone B-producing Strains by Genome Shuffling

Wang Dahong¹, Liu Fei²

(1. College of Food and Bioengineering，Henan University of Science and Technology，Luoyang 471023；2. School of Life Science and Technology，Nanyang Normal University，Nanyang 473061)

Received:2012-11-05 Revised:2013-05-24 Published:2013-05-24 Online:2013-05-24

摘要/Abstract

摘要： 以纤维堆囊菌AHB125经过紫外诱变和2%乙酸钠抗性筛选出来的SC4-14和SC4-56为出发菌株，采用基因组重排技术选育Epothilone B高产菌株，并探索该技术在菌种选育中的基本规律。通过4轮基因组重排成功选育出了5株遗传稳定的Epothilone B高产菌株，其中1株重排菌株F4-28 Epothilone B产量达到67.4 mg/L，是原始菌株纤维堆囊菌AHB125 Epothilone B产量的8倍，是两亲本菌株Epothilone B产量的4-5倍，并且F4-28的发酵周期比SC4-56短1 d。因此，基因组重排技术可有效的选育出Epothilone B高产菌株。

关键词: Epothilone, B, 纤维堆囊菌, AHB125, 基因组重排

Abstract: The mutation of Sorangium cellulosum AHB125 was induced using the classical UV-mutation method plus selection pressures of 2% sodium acetate. Two mutants, SC4-14 and SC4-56, were used as original strains to screen and breed high Epothilone B-producing strains by genome shuffling and explore the basic rule of genome shuffling in strain breeding. Five hereditarily stable strains with high Epothilone B production were obtained after four cycles of genome shuffling. Strain F4-28 yielded 67.4 mg/L of Epothilone B, which was 8-fold higher than that of the initial strain S. cellulosum AHB125 and 4 to 5-fold higher than the mutants used as the original strains for genome shuffling. In addition, the fermentation period of F4-28 was one day shorter than that of SC4-56. The technique of genome shuffling was an efficient method to screen and breed high Epothilone B-producing strains.

Key words: Epothilone, B, Sorangium cellulosum, AHB125, Genome shuffling

王大红, 刘飞. 基因组重排选育Epothilone B高产菌株[J]. 生物技术通报, 2013, 0(5): 155-160.

Wang Dahong, Liu Fei. Breeding of High Epothilone B-producing Strains by Genome Shuffling[J]. Biotechnology Bulletin, 2013, 0(5): 155-160.

参考文献

[1] Gerth K, Bedorf N, Hofle G, et al. Epothilones A and B：antifungal and cytotoxic compounds from Sorangium cellulosum (Myxobacteria)：production, physico-chemical and biological properties [J]. Journal Antibiotics, 1996, 49(3)：560-563.
[2] Whitney H, Juwen D, Quincy Q. The microtubule inhibiting agent epothilone B antagonizes glioma cell motility associated with reorganization of the actin-binding protein α-actinin 4 [J]. Oncology Reports, 2011, 25(3)：887-893.
[3] Kumar A, Heise H, Blommers M, et al. Interaction of Epothilone B (Patupilone) with microtubules as detected by two-dimensional solid-state NMR spectroscopy [J]. Angewandte Chemie, 2010, 49(41)：7504-7507.
[4] Danishefsky S. On the potential of natural products in the discovery of pharma leads：A case for reassessment [J]. Natural Product Reports, 2010, 27：1114-1116.
[5] Harichand-Herdt S, O’Regan RM. Identifying subsets of metastatic breast cancer patients likely to benefit from treatment with the Epothilone B analog Ixabepilone [J]. American Journal of Clinical Oncology, 2010, 33(6)：561-567.
[6] Frykman S, Tsuruta H, Lau J, et al. Modulation of epothilone analog production through media design [J]. Journal of Industrial Microbiology Biotechnology, 2002, 28(1)：17-20.
[7] Zhang YX, Perry KA, Vinci V, et al. Genome shuffling leads to rapid phenotypic improvement in bacteria [J]. Nature, 2002, 6872：644-646.
[8] Gendy M, Bondkly A. Genome shuffling of marine derived bacterium Nocardia sp. ALAA 2000 for improved ayamycin production [J]. Antonie van Leeuwenhoek, 2011, 99(4)：773-780.
[9] Xu F, Jin H, Li H, et al. Genome shuffling of Trichoderma viride for enhanced cellulase production [J]. Annals of Microbiology, 2012, 62(2)：509-515.
[10] 王大红, 原江锋, 郭文杰.产Epothilone B纤维堆囊菌的选育与发酵优化[J].生物技术, 2012, 22(3)：70-72.
[11] Julien B, Shah S. Heterologous expression of epothilone biosynthetic genes in Myxococcus xanthus [J]. Antimicrobial Agents and Chemotherapy, 2002, 46(9)：2772-2778.
[12] Petri R, Claudia SD. Dealing with complexity：evolutionary engineering and genome shuffling [J]. Current Opinion in Biotechnology, 2004, 15(4)：298-304.
[13] 张旭, 张惠文, 徐明恺.基因组重排技术选育乳链菌肽高产菌株[J].中国生物制品学杂志, 2010, 23(10)：1065-1073.
[14] Gong G, Sun X, Liu X, et al. Mutation and a high-throughput screening method for improving the production of Epothilones of Sorangium [J]. Journal of Industrial Microbiology Biotechnology, 2007, 34(9)：615-623.
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