Hernández Christian, Farnet Da Silva Anne-Marie, Ziarelli Fabio, Perraud-Gaime Isabelle, Gutiérrez-Rivera Beatriz, García-Pérez José Antonio, Alarcón Enrique
Instituto de Biotecnología y Ecología Aplicada (INBIOTECA), Universidad Veracruzana, Av. de las Culturas Veracruzanas No. 101, Col. Emiliano Zapata, C.P. 910910, Xalapa, Veracruz, Mexico.
Equipe Systèmes Microbiens, IMBE, UMR CNRS 7263, IRD 237, Aix Marseille Université, Avignon Université, Campus de l'Etoile, Site de St. Jérôme, Marseille, France.
Appl Microbiol Biotechnol. 2017 Feb;101(3):1189-1201. doi: 10.1007/s00253-016-7890-0. Epub 2016 Oct 14.
The use of synthetic dyes for laccase induction in vivo has been scarcely explored. We characterized the effect of adding different synthetic dyes to liquid cultures of Pycnoporus sanguineus on laccase production. We found that carminic acid (CA) can induce 722 % and alizarin yellow 317 % more laccase than control does, and they promoted better fungal biomass development in liquid cultures. Aniline blue and crystal violet did not show such positive effect. CA and alizarin yellow were degraded up to 95 % during P. sanguineus culturing (12 days). With this basis, CA was selected as the best inducer and used to evaluate the induction of laccase on solid-state fermentation (SSF), using sugarcane bagasse (SCB) as substrate, in an attempt to reach selective delignification. We found that laccase induction occurred in SSF, and a slight inhibition of cellulase production was observed when CA was added to the substrate; also, a transformation of SCB under SSF was followed by the C cross polarization magic angle spinning (CPMAS) solid-state nuclear magnetic resonance (NMR). Results showed that P. sanguineus can selectively delignify SCB, decreasing aromatic C compounds by 32.67 % in 16 days; O-alkyl C region (polysaccharides) was degraded less than 2 %; delignification values were not correlated with laccase activities. Cellulose-crystallinity index was increased by 27.24 % in absence of CA and 15.94 % when 0.01 mM of CA was added to SCB; this dye also inhibits the production of fungal biomass in SSF (measured as alkyl C gain). We conclude that CA is a good inducer of laccase in liquid media, and that P. sanguineus is a fungus with high potential for biomass delignification.
合成染料在体内诱导漆酶的应用鲜有研究。我们表征了向血红密孔菌液体培养物中添加不同合成染料对漆酶产生的影响。我们发现,胭脂红酸(CA)能比对照诱导产生多722%的漆酶,茜素黄能比对照诱导产生多317%的漆酶,且它们能促进液体培养物中真菌生物量更好地生长。苯胺蓝和结晶紫未显示出这种积极作用。在血红密孔菌培养(12天)过程中,CA和茜素黄的降解率高达95%。在此基础上,选择CA作为最佳诱导剂,并用于评估其在以甘蔗渣(SCB)为底物的固态发酵(SSF)中对漆酶的诱导作用,以尝试实现选择性脱木质素。我们发现在SSF中发生了漆酶诱导,当将CA添加到底物中时,观察到纤维素酶产生略有抑制;此外,通过交叉极化魔角旋转(CPMAS)固态核磁共振(NMR)跟踪了SSF条件下SCB的转化。结果表明,血红密孔菌能选择性地使SCB脱木质素,在16天内使芳香族C化合物减少32.67%;O-烷基C区域(多糖)的降解率小于2%;脱木质素值与漆酶活性不相关。在不添加CA的情况下,纤维素结晶度指数增加了27.24%,当向SCB中添加0.01 mM CA时,纤维素结晶度指数增加了15.94%;这种染料也抑制了SSF中真菌生物量的产生(以烷基C增加量衡量)。我们得出结论,CA是液体培养基中漆酶的良好诱导剂,且血红密孔菌是一种具有高生物质脱木质素潜力的真菌。