Matselyukh B P, Polishchuk L V, Lukyanchuk V V, Golembiovska S L, Lavrenchuk V Y
Mikrobiol Z. 2016 Nov-Dec;78(6):60-70.
Streptomyces globisporus 1912 and its derivatives 1912-2 and 1912-4Crt are the producers of the landomycin E, carotenoids and the regulator of antibiotics biosynthesis and morphogenesis of streptomycetes. The genome DNA of two mutant strains, 1912-2, the more effective producer of the landomycin E and the regulator, and 1912-4Crt, the producer of beta-carotene and lycopene, was sequenced by Illumina. Comparative analysis of the DNA sequences using GenBank data allowed localization of 36 landomycin E biosynthetic genes lnd of S. globisporus 1912 in one cluster. Twenty of these lnd genes have been sequenced for the first time. The new regulatory responce gene lndRR and the sensor kinase gene lndY1 were proposed as the members of the putative two-component system. High identity (94 – 95 %) was determined for the lnd genes of the 1912 strain and those of the metagenomic clone AZ97. Seven carotenoid biosynthetic genes crt of the strain 1912-4Crt were sequenced and localized in one cluster consisting of two convergent operons from 4 and 3 crt genes. High homology (93 %) of the crt gene clusters of S. globisporus 1912 and S. griseus IFO 13350 was shown. Two non-punctual repeats (NPRs) of 21 bp were identified in the sequence of crtY gene coding lycopene cyclase. It was shown that the deletion of 117 bp including the sequence between NPRs of 96 bp and one NPR from 5`-side activated the crt gene cluster and increased the production of beta carotene (6.91 mg/l) and lycopene (3.24 mg/l) by the strain 1912-4Crt. Deletion of 86 bp was revealed in the regulatory gene lndRR resulting in the deficiency of landomycin E production in the strain 1912-4Crt. The DNA sequences of crt and lnd genes of S. globisporus 1912 were submitted to the NCBI database with accession numbers KM349312 and KJ645792, respectively. S. globisporus 1912 produced a low-molecular-weight compound that, like A-factor, restored the landomycin E and streptomycin biosynthesis and sporulation of the defective mutants S. globisporus 1912-B2 and S. griseus 1439, respectively. The compound was purified by thin layer chromatography and HPLC. It had an absorption maximum at λmax= 245 nm and a molecular mass m/z 244. On the basis of NMR spectroscopy the chemical structure of the transcriptional regulator was elucidated as the new (L)-N-methylphenylalanyl-dehydrobutyrine diketopiperazine.
球形链霉菌1912及其衍生物1912 - 2和1912 - 4Crt是地霉素E、类胡萝卜素的产生菌，也是链霉菌抗生素生物合成及形态发生的调节因子。通过Illumina技术对两个突变株1912 - 2（地霉素E和调节因子的更高效产生菌）和1912 - 4Crt（β-胡萝卜素和番茄红素产生菌）的基因组DNA进行了测序。利用GenBank数据对DNA序列进行比较分析，可将球形链霉菌1912的36个地霉素E生物合成基因lnd定位在一个簇中。其中20个lnd基因首次被测序。新的调节响应基因lndRR和传感器激酶基因lndY1被认为是假定的双组分系统的成员。1912菌株的lnd基因与宏基因组克隆AZ97的lnd基因具有高度同一性（94 - 95%）。对菌株1912 - 4Crt的7个类胡萝卜素生物合成基因crt进行了测序，并定位在一个由4个和3个crt基因组成的两个反向操纵子的簇中。结果表明，球形链霉菌1912和灰色链霉菌IFO 13350的crt基因簇具有高度同源性（93%）。在编码番茄红素环化酶的crtY基因序列中鉴定出两个21 bp的非点突变重复序列（NPRs）。结果表明，缺失包括96 bp的NPRs之间的序列和5`端的一个NPRs在内的117 bp可激活crt基因簇，并使菌株1912 - 4Crt的β-胡萝卜素（6.91 mg/l）和番茄红素（3.24 mg/l）产量增加。在调节基因lndRR中发现缺失86 bp，导致菌株1912 - 4Crt中地霉素E产量不足。球形链霉菌1912的crt和lnd基因的DNA序列已分别提交至NCBI数据库，登录号分别为KM349312和KJ645792。球形链霉菌1912产生一种低分子量化合物，该化合物与A因子类似，分别恢复了缺陷突变株球形链霉菌1912 - B2和灰色链霉菌1439的地霉素E和链霉素生物合成及孢子形成。该化合物通过薄层色谱和HPLC进行纯化。其最大吸收波长为λmax = 245 nm，分子量为m/z 244。基于核磁共振光谱，阐明了该转录调节因子的化学结构为新的（L）-N-甲基苯丙氨酰-脱氢丁氨酸二酮哌嗪。
