Salim Rasha G, Fadel Mohamed, Youssef Yehya A, Taie Hanan A A, Abosereh Nivien A, El-Sayed Ghada M, Marzouk Mohamed
Microbial Genetic Department, Biotechnology Research Institute, National Research Centre, 33 El-Bohouth St. (Former El-Tahrir St.), Dokki, Cairo, 12622, Egypt.
Microbial Chemistry Department, Biotechnology Research Institute, National Research Centre, 33 El-Bohouth St. (Former El-Tahrir St.), Dokki, Cairo, 12622, Egypt.
J Genet Eng Biotechnol. 2022 Apr 22;20(1):62. doi: 10.1186/s43141-022-00335-2.
During the last decade, enormous research efforts have been directed at identifying potent microorganisms as sustainable green cell factories for eco-friendly pigments. Talaromyces atroroseus has recently been shown to excrete large amounts of azaphilone mycotoxin-free red pigment mixture comprising some known coloring components together with many uncharacterized metabolites. In this study, a new Talaromyces atroroseus isolate was identified via sequencing of the fragment of the nuclear ribosomal gene cluster containing internal transcribed spacers and 5.8S rRNA gene. The parameters that affected the level of pigment production were optimized in uncommon static conditions of culture and genetic improvement, via γ-irradiation, to improve pigment yield. Moreover, chemical characterization using LC/MS and skin safety test of the target pigment mixture were precisely conducted to maximize its benefits as a natural and safe red pigment for wool fabrics.
Molecular identification via the sequencing of the ITS of the rDNA encoding gene cluster revealed that the fungal isolate TRP-NRC was T. atroroseus TRP-NRC (deposited in GenBank under accession number MW282329). In the static conditions of culture, pigment production was dramatically enhanced to 27.36 g/L in an optimum yeast malt peptone medium of 2% mannitol at pH 2-4.5 and 30 °C for 7 days of incubation. Under exposure to a 400-Gy γ-radiation dose, pigment yield was increased to a 3-fold level higher than that recorded for the wild type. Based on the inter-simple sequence repeats (ISSR), as a molecular marker tool, the wild-type T. atroroseus TRP-NRC strain and its mutants were discriminated. The UHPLC/HRESI-MS analytical tool characterized 60 metabolites, including many unknown molecules, at appropriate concentrations. It is worthy to note that four mitorubrin derivatives were identified for the first time in T. atroroseus, i.e., mitorubrinolamine acetate, dihydro-PP-O, mitorobrinal, and mitorubrinol. The range of irritation indexes (0-0.1) demonstrated an adequate skin safety after the direct local application of the pigment mixture. Finally, the pigment mixture exhibited a remarkably good dyeing ability in wool fabrics, with high-fastness properties.
Because of its sustainable and economic production, the target red pigment mixture may be applied in the future in textile, food, cosmetics, or different pharmaceutical industries after extensive conventional safety and toxicity studies, which are currently under consideration.
在过去十年中,人们投入了大量研究精力来寻找强大的微生物,将其作为生产环保型色素的可持续绿色细胞工厂。最近发现,黑玫瑰色篮状菌能分泌大量不含氮杂蒽酮霉菌毒素的红色色素混合物,其中包括一些已知的色素成分以及许多未鉴定的代谢产物。在本研究中,通过对包含内部转录间隔区和5.8S rRNA基因的核糖体基因簇片段进行测序,鉴定出一种新的黑玫瑰色篮状菌分离株。在不常见的静态培养条件下,通过γ射线辐照对影响色素产量的参数进行优化,以提高色素产量。此外,还精确地进行了使用液相色谱/质谱联用仪的化学表征以及目标色素混合物的皮肤安全性测试,以最大限度地发挥其作为羊毛织物天然安全红色色素的优势。
通过对编码基因簇的rDNA的ITS进行测序,分子鉴定表明真菌分离株TRP-NRC为黑玫瑰色篮状菌TRP-NRC(保藏于GenBank,登录号为MW282329)。在静态培养条件下,在含有2%甘露醇、pH值为2 - 4.5、30℃的最佳酵母麦芽蛋白胨培养基中培养7天,色素产量显著提高至27.36 g/L。在400 Gy的γ射线辐照剂量下,色素产量比野生型提高了3倍。基于简单重复序列区间(ISSR)这一分子标记工具,区分了野生型黑玫瑰色篮状菌TRP-NRC菌株及其突变体。超高效液相色谱/高分辨电喷雾电离质谱联用分析工具鉴定出60种代谢产物,包括许多未知分子,且浓度适宜。值得注意的是,在黑玫瑰色篮状菌中首次鉴定出四种米托菌素衍生物,即米托菌素醇胺乙酸酯、二氢-PP-O、米托菌素醛和米托菌素醇。刺激指数范围为0 - 0.1,表明直接局部应用色素混合物后皮肤安全性良好。最后,色素混合物在羊毛织物上表现出显著良好的染色能力,且色牢度高。
由于其可持续且经济的生产方式,在目前正在进行的广泛常规安全性和毒性研究之后,目标红色色素混合物未来可能应用于纺织、食品、化妆品或不同的制药行业。