Characterization of a G protein α subunit encoded gene from the dimorphic fungus-Tremella fuciformis

Antonie van Leeuwenhoek - Tremella fuciformis is a dimorphic fungus which can undertake the reversible transition between yeast and pseudohypha forms. G protein α subunit (Gα) carries...     

微载体培养中不同感染剂量的麻疹病毒对病毒产量的影响

用微载体培养法在搅拌瓶和生物反应罐中小规模培养麻疹病毒,以探索在可能实现的大规模工业化麻疹生产中不同感染剂量ＭｕｌｔｉｐｌｙｏｆＩｎｆｅｃｔｉｏｎ（ｍ．ｏ．ｉ）的麻疹病毒对病毒产量的影响。增加ｍ．ｏ．ｉ可以将病毒的高峰期提前两到三天,但病毒产量却并不因此而提高,还有可能引起微载体上贴附的细胞层损坏加剧,从而被迫过早终止培养,导致病毒总产量下降。     

Use of the yeast-like cells of <Emphasis Type="Italic">Tremella fuciformis</Emphasis> as a cell factory to produce a <Emphasis Type="Italic">Pleurotus ostreatus</Emphasis> hydrophobin

Objectives

To obtain hydrophobin, a Class I hydrophobin gene, Po.hyd from Pleurotus ostreatus, was transformed into the yeast-like cells of Tremella fuciformis using Agrobacterium tumefaciens.

Results

The hydrophobin Po.HYD from P. ostreatus was heterogeneously expressed by the yeast-like cells of T. fuciformis. Plasmids harboring the Po.hyd gene driven by endogenous glyceraldehyde-3-phosphate dehydrogenase promoter were transformed by A. tumefaciens. The integration and expression of the rPo.HYD in the T. fuciformis cells were confirmed by PCR, Southern blot, fluorescence microscopy and quantitative real-time PCR. SDS-PAGE demonstrated that the rPo.HYD was extracted with the expected MW of 14 kDa. The yield of purified rPo.HYD was 0.58 mg/g dry wt. The protein, with its ability to stabilize oil droplets, exhibited a better emulsifying activity than the typical food emulsifiers Tween 20 and sodium caseinate.

Conclusion

Tremella fuciformis can be used as a cell factory to produce hydrophobin on a large scale for the food industry.

     

Comparing the sugar profiles and primary structures of alkali-extracted water-soluble polysaccharides in cell wall between the yeast and mycelial phases from <Emphasis Type="Italic">Tremella fuciformis</Emphasis>

To gain insights into dimorphism, cell wall polysaccharides from Tremella fuciformis strains were obtained from alkali-extracted water-soluble fractions PTF-M38 (from the mycelial form), PTF-Y3 and PTF-Y8 (from the yeast form) of T. fuciformis strains were used to gain some insights into dimorphism study. Their chemical properties and structural features were investigated using gel permeation chromatography, gas chromatography, UV and IR spectrophotometry and Congo red binding reactions. The results indicated that the backbones of PTF-M38, PTF-Y3 and PTF-Y8 were configured with α-linkages with average molecular weights of 1.24, 1.08, and 1.19 kDa, respectively. PTF-M38 was mainly composed of xylose, mannose, glucose, and galactose in a ratio of 1:1.47:0.48:0.34, while PTF-Y3 and PTF-Y8 were mainly composed of xylose, mannose and glucose in a ratio of 1:1.65:4.06 and 1:1.21:0.44, respectively. The sugar profiles of PTF-M38, PTF-Y3 and PTF-Y8 were also established for further comparison. These profiles showed that all three polysaccharides contained the same sugars but in different ratios, and the carbon sources (xylose, mannose, glucose, and galactose) affected the sugar ratios within the polysaccharides.