Capability of a large bacterial artificial chromosome clone harboring multiple biosynthetic gene clusters for the production of diverse compounds

IF 2.1 4区医学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of Antibiotics Pub Date : 2024-03-04 DOI:10.1038/s41429-024-00711-9

Kei Kudo, Takehiro Nishimura, Miho Izumikawa, Ikuko Kozone, Junko Hashimoto, Manabu Fujie, Hikaru Suenaga, Haruo Ikeda, Nori Satoh, Kazuo Shin-ya

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Abstract

The biosynthetic gene clusters (BGCs) for the macrocyclic lactone-based polyketide compounds are extremely large-sized because the polyketide synthases that generate the polyketide chains of the basic backbone are of very high molecular weight. In developing a heterologous expression system for the large BGCs amenable to the production of such natural products, we selected concanamycin as an appropriate target. We obtained a bacterial artificial chromosome (BAC) clone with a 211-kb insert harboring the entire BGC responsible for the biosynthesis of concanamycin. Heterologous expression of this clone in a host strain, Streptomyces avermitilis SUKA32, permitted the production of concanamycin, as well as that of two additional aromatic polyketides. Structural elucidation identified these additional products as ent-gephyromycin and a novel compound that was designated JBIR-157. We describe herein sequencing and expression studies performed on these BGCs, demonstrating the utility of large BAC clones for the heterologous expression of cryptic or near-silent loci.

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携带多个生物合成基因簇的大型细菌人工染色体克隆生产多种化合物的能力。

大环内酯类多酮化合物的生物合成基因簇（BGCs）非常大，因为产生基本骨架多酮链的多酮合成酶分子量非常高。在开发适合生产此类天然产物的大型 BGCs 的异源表达系统时，我们选择了康加霉素作为合适的目标。我们获得了一个细菌人工染色体（BAC）克隆，该克隆具有 211 kb 的插入片段，其中包含负责生物合成 concanamycin 的整个 BGC。将该克隆异源表达于宿主菌株--阿维米蒂斯链霉菌 SUKA32--中，不仅能生产出凹霉素，还能生产出另外两种芳香族多酮类化合物。通过结构阐释，我们确定这些额外的产物为 ent-gephyromycin 和一种新型化合物，并将其命名为 JBIR-157。我们在此描述了对这些 BGC 进行的测序和表达研究，证明了大型 BAC 克隆在隐性或接近沉默基因座的异源表达方面的实用性。

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来源期刊

Journal of Antibiotics 医学-免疫学

CiteScore

6.60

自引率

3.00%

发文量

审稿时长

1 months

期刊介绍： The Journal of Antibiotics seeks to promote research on antibiotics and related types of biologically active substances and publishes Articles, Review Articles, Brief Communication, Correspondence and other specially commissioned reports. The Journal of Antibiotics accepts papers on biochemical, chemical, microbiological and pharmacological studies. However, studies regarding human therapy do not fall under the journal’s scope. Contributions regarding recently discovered antibiotics and biologically active microbial products are particularly encouraged. Topics of particular interest within the journal''s scope include, but are not limited to, those listed below: Discovery of new antibiotics and related types of biologically active substances Production, isolation, characterization, structural elucidation, chemical synthesis and derivatization, biological activities, mechanisms of action, and structure-activity relationships of antibiotics and related types of biologically active substances Biosynthesis, bioconversion, taxonomy and genetic studies on producing microorganisms, as well as improvement of production of antibiotics and related types of biologically active substances Novel physical, chemical, biochemical, microbiological or pharmacological methods for detection, assay, determination, structural elucidation and evaluation of antibiotics and related types of biologically active substances Newly found properties, mechanisms of action and resistance-development of antibiotics and related types of biologically active substances.