Matselyukh B P, Matselyukh D Ya, Golembiovska S L, Polishchuk L V, Lavrinchuk V Ya
Zabolotny Institute of Microbiology and Virology, National Academy of Sciences of Ukraine, 154 Akad Zabolotny St., Kyiv D03680, Ukraine.
Mikrobiol Z. 2013 Nov-Dec;75(6):10-6.
Hyperpigmented mutants of Streptomyces globisporus 1912-Hp7 and Blakeslea trispora 18(+), 184(-) were isolated by action of hydrogen peroxide and nitrosoguanidine, correspondingly, from initial strains S. globisporus 1912-4Lcp and B. trispora 72(-), 198(+). The carotenoids of dry biomass of obtained strains, rubbed thoroughly with glass powder by a pestle in porcelain mortar were extracted by acetone and purified by TLC. Identification of the individual carotenoids was performed by means of HPLC and LC/MS spectrometry. It was shown that strain S. globisporus 1912-4Crt produced beta-carotene/lycopene (6.91/3.24 mg/L), mutants 1912-4Lcp and 1912-7Hp synthesized only lycopene (26.05 and 50.9 mg/L, respectively), and strains B. trispora 18(+) and 184(-)-beta-carotene (6.2% in dry biomass or more 2.5 g/L) without illumination in shake flasks. It is the first example of high constitutive production of the carotenoids by the representative of genus Streptomyces without photoinduction or increased synthesis of sigma factor The improved strains of B. trispora 18(+) and 184(-) can be used for biotechnological production of beta-carotene in industrial conditions.
分别通过过氧化氢和亚硝基胍的作用，从初始菌株球形链霉菌1912 - 4Lcp和三孢布拉氏霉72(-)、198(+)中分离得到了球形链霉菌1912 - Hp7和三孢布拉氏霉18(+)、184(-)的色素沉着突变体。将所得菌株的干燥生物量与玻璃粉在瓷研钵中用杵充分研磨后，用丙酮提取类胡萝卜素，并通过薄层层析进行纯化。通过高效液相色谱和液相色谱/质谱联用对各个类胡萝卜素进行鉴定。结果表明，球形链霉菌1912 - 4Crt菌株产生β-胡萝卜素/番茄红素（6.91/3.24毫克/升），突变体1912 - 4Lcp和1912 - 7Hp仅合成番茄红素（分别为26.05和50.9毫克/升），而三孢布拉氏霉18(+)和184(-)菌株在摇瓶中无光条件下产生β-胡萝卜素（干生物量中占6.2%或超过2.5克/升）。这是链霉菌属代表菌株在没有光诱导或增加σ因子合成的情况下高水平组成型生产类胡萝卜素的首个实例。改良后的三孢布拉氏霉18(+)和184(-)菌株可用于工业条件下β-胡萝卜素的生物技术生产。
