Improvement of FR901379 production by mutant selection and medium optimization
Improvement of FR901379 production by mutant selection and medium optimization
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DOI：
10.1016/j.jbiosc.2009.01.002
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发表时间：
2009-05-01
影响因子：
2.8
通讯作者：
Honda, Hiroyuki
中科院分区：
文献类型：
--
作者：
Kanda, Munekazu;Tsuboi, Masaru;Honda, Hiroyuki
FR 901379（WF 11899 A）是一种新的棘白菌素型脂肽抗生素，由Coleophoma empetri F-11899产生。米卡芬净（FK 463）来源于脱酰FR 901379的化学修饰。本论文从野生型开始，进行了7代的品系选育。设计了用于筛选的选择培养基和用于高FR 901379产量的生产培养基。筛选平板中氯化钠含量对FR 901379的产量有影响，高产菌株的殖民地缩小。选择白念珠菌琼脂片法中菌落收缩率大、抑菌圈大的菌株作为筛选标记。在高氯化钠CMA培养基中，突变株M-1 ~ M-3的FR 901379产量较高，其中M-3的产量为5.0 U/mL，而野生型菌株的产量为1.0 U/mL。在A-2培养基中添加6%的可溶性淀粉作为碳源，0.6%的硫酸铵作为氮源，对突变体的筛选也是有效的。突变体M-4（第4代）的FR 901379产量增加到16.0 U/mL。培养基中磷酸盐的浓度似乎抑制生长，从而延长培养期。当设计并使用补充有低浓度磷酸盐和硫酸镁作为硫酸盐源的A-3培养基时，连续获得了具有提高的产量的突变体。最后，变异菌株M-7显示出30.0 U/mL的产量，这是约30倍的野生型菌株。(c)2009年，生物技术学会，日本。All rights reserved.
FR901379 (WF11899A) is a novel echinocandin type of lipopeptide antibiotic produced by Coleophoma empetri F-11899. Micafungin (FK463) is derived from the chemical modification of deacylated FR901379. In the present paper, we performed seven generation's strain-breeding, beginning with a wild type, was performed. Selection medium for screening and production medium for high FR901379 production were designed. Sodium chloride content in the selection plate was affected to FR901379 production and shrinkage of the colony size was observed in high producing strains. As selection markers, large colony-shrinking rate and large inhibition circle in the agar-piece method using C albicans was selected. Using CMA medium with high sodium chloride, 3 mutants, M-1 to M-3, have achieved a high FR901379 production and M-3 showed 5.0 U/mL,, while 1.0 U/mL, of production was achieved in wild type strain. A-2 medium supplemented with 6% of soluble starch as a carbon source and 0.6% of ammonium sulfate as nitrogen source was also further effective for mutant screening. The FR901379 production of mutant M-4 (fourth generation) increased until 16.0 U/mL The concentration of the phosphate salt in the medium seemed to inhibit the growth so as to extend the culture period. When the A-3 medium supplemented with low concentration of phosphate salt and magnesium sulfate as a sulfate source was designed and used, mutants with improved production were successively obtained. Finally, variant strain M-7 showed 30.0 U/mL of production, which was about 30 times higher than that of the wild strain. (c) 2009, The Society for Biotechnology, Japan. All rights reserved.