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首页> 外文期刊>RSC Advances >Effect of overexpression of endogenous and exogenous Streptomyces antibiotic regulatory proteins on tacrolimus (FK506) production in Streptomyces sp KCCM11116P
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Effect of overexpression of endogenous and exogenous Streptomyces antibiotic regulatory proteins on tacrolimus (FK506) production in Streptomyces sp KCCM11116P

机译：内源性和外源链霉菌抗生素调控蛋白在链霉菌SP KCCM11116P中产生抗生素调节蛋白的影响

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摘要

With the development of synthetic biology and systems biology, overproduction of biomolecules can be achieved by manipulation of regulatory proteins from various sources. FK506 (tacrolimus) is a 23-membered macrolide antibiotic and an important immunosuppressant. Overproduction of FK506 has been achieved by the manipulation of regulatory genes that are located inside the biosynthetic gene cluster, however, little is known on the effect of other regulators on FK506 production. In this study, the effects of three Streptomyces antibiotic regulatory proteins (SARPs) on FK506 production were investigated. These SARPs include bulZ and bulY that are cloned from a FK506 producer Streptomyces sp. KCCM11116P, and one novel SARP family regulatory protein Sx5140 that was obtained from a marine streptomycete S. xinghaiensis. The production titer of the engineered strains exhibited a higher production level compared to that in the control strain (227.99 mg L-1), and overexpression of bulZ resulted in the highest FK506 titer (365.59 mg L-1), which is 1.6-fold of that of the control. Real-time PCR analysis showed that overexpression of bulZ and bulY resulted in increased transcription of tsuR1 which is the gamma-butyrolactone receptor. Variation of transcription levels of fkbN, the positive regulator of FK506 biosynthesis, as well as fkbG, fkbH, fkbI, tcsA, tcsB, tcsD and fkbQ genes that are involved in the FK506 biosynthesis were observed in the SARP overexpression strains compared to those in the control strain. Our results demonstrate the promising potential to utilize alternative regulatory proteins including both endogenous and exogenous ones to enhance the production of useful compounds in microbial strains.
机译：随着合成生物学和系统生物学的发展，通过操纵来自各种来源的调节蛋白，可以实现生物分子的过度生产。 FK506（Tacrolimus）是23元大环内酯抗生素和重要的免疫抑制剂。然而，通过操纵位于生物合成基因簇内的调节基因来实现FK506的过度生产，然而，对其他调节剂对FK506生产的影响很少。在这项研究中，研究了三种链霉菌抗生素调节蛋白（SARP）对FK506生产的影响。这些SARP包括来自FK506生产商链霉菌SP的FK506生产商克隆的山谷和粗壮。 KCCM11116P和一项新的SARP家庭调节蛋白SX5140是从海洋链霉素S. Xinghaiensis获得的。与对照菌株（227.99mg L-1）中的制造滴度相比，工程株的生产滴度表现出更高的生产水平，并且膨胀的过表达导致最高的FK506滴度（365.59mg L-1），为1.6倍对照的影响。实时PCR分析表明，膨胀和茂硼的过度表达导致TSUR1的转录增加，这是γ-丁内酯受体。 FKBN的转录水平，FK506生物合成的阳性调节剂，以及参与FK506生物合成的FKBG，FKBH，FKBI，TCSA，TCSB，TCSD和FKBQ基因，与其中的SARP过表达菌株相比观察到控制菌株。我们的结果证明了利用包括内源和外源性的替代调节蛋白的有希望的潜力，以增强微生物菌株中的有用化合物的产生。

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  • 来源
    《RSC Advances》 |2015年第21期|共7页
  • 作者

    Chen Chao; Zhao Xinqing; Chen Liangyu; Jin Yingyu; Zhao Zongbao K.; Suh Joo-Won;

  • 作者单位

    Dalian Univ Technol Sch Life Sci &

    Biotechnol Dalian 116024 Peoples R China;

    Dalian Univ Technol Sch Life Sci &

    Biotechnol Dalian 116024 Peoples R China;

    Dalian Univ Technol Sch Life Sci &

    Biotechnol Dalian 116024 Peoples R China;

    Myongji Univ Div Biosci &

    Bioinformat Yongin 449728 South Korea;

    Chinese Acad Sci Dept Biotechnol Dalian Inst Chem Phys Dalian 116023 Peoples R China;

    Myongji Univ Div Biosci &

    Bioinformat Yongin 449728 South Korea;

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