HETEROLOGOUS EXPRESSION AND CHARACTERIZATION OF MANNITOL-1-PHOSPHATE DEHYDROGENASE FROM THE BASIODIOMYCETE PHOLIOTA NAMEKO

ABSTRACT The gene encoding mannitol-1-phosphate dehydrogenase, mpd, which has similar function with alcohol dehydrogenase, has been sequenced and characterized from the basiodiomycete Pholiota nameko. The coding region of mpd was composed of 2992 bp and found to encode a polypeptide of 359 amino acids that has similarity with Laccaria bicolor. To evaluate the expression level of mannitol-1-phosphate from P. nameko, mpd cDNA was inserted into pCold shock vector and expressed in host BL21 (DE3) by 24 h induction with 1 mM isopropyl 1-thio-β-Dgalactopyranoside at 15 O C after 2 h growth at 37 O C. The purified protein was detected by SDS-PAGE and western blot

Mycelial Growth-promoting Potential of Extracellular Metabolites of Paraburkholderia spp. Isolated from Rhizopogon roseolus Sporocarp

This study aimed to investigate the effect of potential metabolite(s) produced by Paraburkholderia spp. isolated from the Rhizopogon roseolus (shouro mushroom) sporocarp on the mycelial growth of R. roseolus. For this purpose, we selected two molecularly identified bacteria: P. fungorum GIB024 and P. caledonica KN1. Direct confrontation assay at three different distances, a pour plate method that sampled bacterial spent broth either with and without agitation at 25 °C, and an indirect confrontation assay was carried out in order to assess the R. roseolus growth-promoting ability of Paraburkholderia spp. These assessments were carried out in a 1:5 diluted Melin-Norkran-modified medium with glucose (hs-dMMN) and without glucose (ls-dMMN). GIB024 promoted the growth of R. roseolus in ls-dMMN in short distance, whereas KN1 inhibited the growth of the fungus in that condition. In hs-dMMN, both bacteria have neutral or slightly promotion effect toward R. roseolus. We determined from the spent broth analysis that Paraburkholderia spp. that grew axenically under static conditions had a more pronounced mycelial growth-promoting effect on R. roseolus than under agitation conditions. We also found that high concentration of spent broth resulted in a decrease in mycelial growth-promoting ability. Volatile metabolite(s) produced by both bacteria did not promote the mycelial growth of R. roseolus. In conclusion, Paraburkholderia spp. exhibited a species- and nutrient (sugar)-dependent ability to promote the mycelial growth of R. roseolus, and the bacterial soluble metabolite(s) play a crucial role in their growth-promoting ability

Rapid Species Identification of Cooked Poisonous Mushrooms by Using Real-Time PCR▿

Species-specific identification of the major cooked and fresh poisonous mushrooms in Japan was performed using a real-time PCR system. Specific fluorescence signals were detected, and no nonspecific signals were detected. Therefore, we succeeded in developing a species-specific test for the identification of poisonous mushrooms within 1.5 h