ABSTRACT
Lanosterol synthase( LS) is a key enzyme involving in the mevalonate pathway( MVA pathway) to produce lanosterol,which is a precursor of ganoderma triterpenoid. And the transcriptional regulation of LS gene directly affects the content of triterpenes in Ganoderma lucidum. In order to study the transcriptional regulation mechanism of LS gene,yeast one-hybrid technique was used to screen the transcription regulators which interact withthe promoter of LS. The bait vector was constructed by LS promoter,then the vector was transformed yeast cells to construct bait yeast strain. One-hybrid c DNA library was constructed via SMART technology. Then the c DNA and p GADT7-Rec vector were co-transformed into the bait yeast strain to screen the upstream regulatory factors of the promoter region of LS by homologous recombination. Total of 23 positive clones were screened. After sequencing,blast was performed against the whole-genome sequence of G. lucidum. As a result,8 regulatory factors were screened out including the transcription initiation TFIIB,the alpha/beta hydrolase super family,ALDH-SF superfamily,60 S ribosomal protein L21,ATP synthase β-subunit,microtubule associated protein Cript,prote asome subunit β-1,and transaldolase. Until now,the regulation effect of these 8 regulatory factors in G.lucidum has not been reported. This study provides candidate proteins for in-depth study on the expression regulation of LS.
Subject(s)
Gene Library , Intramolecular Transferases/metabolism , Reishi/genetics , Saccharomyces cerevisiae , Transcription Factors/metabolismABSTRACT
El hongo Ganoderma lucidum, en los constituyentes de su biomasa, tiene compuestos con propiedades benéficas para la salud; es por esto que el conocimiento de las condiciones nutricionales adecuadas para su crecimiento permitirá su producción industrial y a bajo costo. En este trabajo se evaluó a nivel de matraz el efecto de la relación C/N, y la presencia de diferentes fuentes de carbono, nitrógeno y micronutrientes sobre la producción de biomasa. Empleando glucosa y peptona como fuentes de carbono y nitrógeno, respectivamente, se encontró una relación C/N óptima de 16,7:1 para la cual la máxima producción de biomasa fue de 25 g/L. Manteniendo esta relación C/N, y sustituyendo la glucosa por lactosa o harina de cebada y la peptona por extracto de levadura, la producción de biomasa se incrementó a 35 g/L. En presencia de harina de cebada la adición al medio de cultivo de sales de Mg y K, y de tiamina, no generó un mayor incremento en la producción de biomasa. La producción de biomasa de G. lucidum se ve favorecida por la presencia en el medio de cultivo de relaciones C/N cercanas a las reportadas conforme a la composición típica de los hongos, así como por la presencia de sustratos complejos como la harina de cebada que le aportan además de la fuente de carbono micronutrientes necesarios para su desarrollo.
Ganoderma lucidum fungus has some biomass components with beneficial health properties. The knowledge about its nutritionals requirements for growing will favor its industrial production at lower cost. In this work, the effect of C/N ratio, the presence of different carbon, nitrogen and micronutrients sources, on fungal biomass production, were evaluated. Using glucose and peptone as carbon and nitrogen sources, respectively, an optimal C/N ratio of 16,7:1 was found, for which the maximal biomass production was 25 g/L. Replacing glucose by lactose or barley flour and peptone by yeast extract at the same C/N ratio, the biomass production was enhanced to 35 g/L. With barley flour in the culture medium, the presence of Mg and K salts and thiamine did not turn out into a major increase of biomass. The G. lucidum biomass production is promoted by C/N ratios in the culture medium nearly equivalent to that found in the fungus, as well as the presence of complex substrates as barley flour which, additionally, contributes with important micronutrients along with the carbon source.