Lipid production from sweet sorghum bagasse through yeast fermentation

Cryptococcus curvatus has great potential in fermenting unconditioned hydrolysates of sweet sorghum bagasse. With hydrolysates obtained by enzymatic hydrolysis of the solid pretreated by microwave with lime, the maximal yeast cell dry weight and lipid content were 10.83 g/l and 73.26%, respectively. For hydrolysates obtained in the same way but without lime, these two parameters were 15.50 g/l and 63.98%, respectively. During yeast fermentation, glucose and xylose were consumed simultaneously while cellobiose was released from the residual bagasse. The presence of lime, on one hand, made cellulose more accessible to enzymes as evidenced by higher total reducing sugar release compared to that without during enzymatic hydrolysis step; on the other hand, it caused the degradation of sugars to non-sugar chemicals during pretreatment step. As a result, higher lipid yield of 0.11 g/g bagasse or 0.65 ton/hectare of land was achieved from the pathway of microwave pretreatment and enzymatic hydrolysis while 0.09 g/g bagasse or 0.51 ton/hectare of land was attained from the process of lime-assisted microwave pretreatment followed by the same enzymatic saccharification.     

A nuclear-encoded intein in the fungal pathogen Cryptococcus neoformans.

We have used comparative sequence analysis to identify an intein-like sequence (protein splicing element) present in Cryptococcus neoformans, a fungal pathogen of humans. The sequence encoding this element is present in the C. neoformans PRP8 gene, as an in-frame insertion relative to the PRP8 genes of other organisms. It contains sequences similar to those of the protein-splicing domains of two previously described yeast inteins (in Saccharomyces cerevisiae and Candida tropicalis), although it lacks any recognizable internal endonuclease domain. The Cryptococcus neoformans intein (Cne PRP8) is only the second to be found in a eukaryote nuclear genome; the previously described yeast inteins occur at the same site in the VMA gene homologues of S. cerevisiae and C. tropicalis. The host gene of the Cryptococcus intein, PRP8, encodes a highly conserved mRNA splicing protein found as part of the spliceosome. The Cne PRP8 intein may be a useful drug target in addressing the cryptococcal infections so prevalent in AIDS patients.     

Control of postharvest<Emphasis Type="Italic"> Rhizopus</Emphasis> rot of peach by microwave treatment and yeast antagonist

The potential of using microwave power alone, or in combination with antagonistic yeast, for control of Rhizopus stolonifer on peach, and its effects on the postharvest quality of peach were investigated. In in vitro tests, the growth of R. stolonifer was completely inhibited by a 2,450 MHz microwave heating for 2 min or more. The population density of R. stolonifer in surface wounds of microwave treatment fruits was significantly lower than that in the control. In vivo studies of inoculation of peach fruit with R. stolonifer followed by microwave treatment, Cryptococcus laurentii or both of them, microwave power and C. laurentii, as stand-alone treatments, were capable of reducing the percentage of decayed fruit from 95% in control, untreated fruit to 42.1% and 75%, respectively. However, in fruit treated with a combination of microwave power and C. laurentii, the percentage of decayed fruits was only 23.7%. The experiments on reducing natural decay development of fruit gave similar results. The microwave treatment, C. laurentii, or both of them did not impair the quality parameters of the fruit. Thus, the combination of microwave and C. laurentii could be an alternative to chemicals for control of postharvest Rhizopus rot on peach fruits.     

The diversity of retrotransposons in the yeast Cryptococcus neoformans.

We have undertaken an analysis of the retrotransposons in the medically important basidiomycetous fungus Cryptococcus neoformans. Using the data generated by a C. neoformans genome sequencing project at the Stanford Genome Technology Center, 15 distinct families of LTR retrotransposons and several families of non-LTR retrotransposons were identified. Members of at least seven families have transposed recently and are probably still active. For several families, only partial elements could be identified and these are quite diverse in sequence, suggesting that they are ancient components of the C. neoformans genome. Most C. neoformans elements are not closely related to previously identified fungal retrotransposons, suggesting that the diversity of fungal retrotransposons has been only sparsely sampled to date. C. neoformans has fewer distinct retrotransposon families than Candida albicans (37 or more), in particular fewer families represented solely by ancient and inactive elements, but it has considerably more families than either Saccharomyces cerevisiae (five) or Schizosaccharomyces pombe (two). The findings suggest that elimination of retrotransposons is faster in C. neoformans than in C. albicans, but perhaps not as rapid as in S. cerevisiae or Sz. pombe. The identification of the retrotransposons of C. neoformans should assist in the molecular characterization of this important pathogen, and also further our understanding of the role played by retroelements in genome evolution.     

Suppression of postharvest blue mould of apple fruit by Cryptococcus laurentii and N6‐benzyladenine

BACKGROUND: Cryptococcus laurentii is a well‐known postharvest yeast antagonist. N⁶‐benzyladenine (6‐BA), a cytokinin plant hormone, has a role in retarding ripening and senescence of harvested produce. This study was conducted to evaluate the efficacy of C. laurentii and 6‐BA in reducing the blue mould disease of apple fruit. RESULTS: The combination of C. laurentii with 6‐BA (20 µg mL^?1) was more effective in suppressing the Penicillium expansum infection in apple fruit wounds than C. laurentii alone, although 6‐BA (20 µg mL^?1) alone neither affected the growth of C. laurentii nor reduced the incidence of the blue mould disease in vivo. Moreover, treatment of apple fruit with C. laurentii and 6‐BA (20 µg mL^?1) resulted in stimulation of superoxide dismutase activity but in inhibition of the increase in peroxidase activity. CONCLUSION: 6‐BA (20 µg mL^?1) could enhance the efficacy of C. laurentii in reducing the postharvest blue mould disease of apple fruit, which offered great potential in minimizing the postharvest decay of apple fruit in an integrated pest management strategy. Copyright © 2008 Society of Chemical Industry     

基于新一代高通量测序技术分析拮抗酵母Cryptoccocus laurentii诱导处理樱桃番茄果实后转录组学的变化

罗伦隐球酵母（Cryptococcus laurentii）作为生物激发子,能显著诱导采后樱桃番茄对灰葡萄孢（Botrytis cinerea）的抗性响应,但其防治机制尚未完全明确。本实验以樱桃番茄为材料,利用新一代高通量测序技术Illumina HiSeqTM4000对罗伦隐球酵母处理及水对照处理的番茄果肉组织进行转录组测序,采用生物信息学方法对差异基因进行功能注释和通路分析,并利用实时荧光定量聚合酶链式反应（real time-quantitative polymerase chain reaction,RT-qPCR）技术对随机选取的差异表达基因进行验证。结果表明,樱桃番茄果实经过罗伦隐球酵母诱导处理48?h后,共有3?141?个基因差异表达,上、下调表达基因分别为1?689?个和1?452?个,这些差异表达基因涉及多个代谢路径,揭示罗伦隐球酵母对番茄的诱导作用是一个多代谢路径参与的调控过程。此外,RT-qPCR验证结果与转录组分析结果趋势一致。     

In Silico Structural Modeling and Analysis of Interactions of Tremellomycetes Cytochrome P450 Monooxygenases CYP51s with Substrates and Azoles

Cytochrome P450 monooxygenase CYP51 (sterol 14α-demethylase) is a well-known target of the azole drug fluconazole for treating cryptococcosis, a life-threatening fungal infection in immune-compromised patients in poor countries. Studies indicate that mutations in CYP51 confer fluconazole resistance on cryptococcal species. Despite the importance of CYP51 in these species, few studies on the structural analysis of CYP51 and its interactions with different azole drugs have been reported. We therefore performed in silico structural analysis of 11 CYP51s from cryptococcal species and other Tremellomycetes. Interactions of 11 CYP51s with nine ligands (three substrates and six azoles) performed by Rosetta docking using 10,000 combinations for each of the CYP51-ligand complex (11 CYP51s × 9 ligands = 99 complexes) and hierarchical agglomerative clustering were used for selecting the complexes. A web application for visualization of CYP51s’ interactions with ligands was developed (http://bioshell.pl/azoledocking/). The study results indicated that Tremellomycetes CYP51s have a high preference for itraconazole, corroborating the in vitro effectiveness of itraconazole compared to fluconazole. Amino acids interacting with different ligands were found to be conserved across CYP51s, indicating that the procedure employed in this study is accurate and can be automated for studying P450-ligand interactions to cater for the growing number of P450s.     

Nitrate Capture Investigation in Plasma-Activated Water and Its Antifungal Effect on Cryptococcus pseudolongus Cells

This research investigated the capture of nitrate by magnesium ions in plasma-activated water (PAW) and its antifungal effect on the cell viability of the newly emerged mushroom pathogen Cryptococcus pseudolongus. Optical emission spectra of the plasma jet exhibited several emission bands attributable to plasma-generated reactive oxygen and nitrogen species. The plasma was injected directly into deionized water (DW) with and without an immersed magnesium block. Plasma treatment of DW produced acidic PAW. However, plasma-activated magnesium water (PA-Mg-W) tended to be neutralized due to the reduction in plasma-generated hydrogen ions by electrons released from the zero-valent magnesium. Optical absorption and Raman spectra confirmed that nitrate ions were the dominant reactive species in the PAW and PA-Mg-W. Nitrate had a concentration-dependent antifungal effect on the tested fungal cells. We observed that the free nitrate content could be controlled to be lower in the PA-Mg-W than in the PAW due to the formation of nitrate salts by the magnesium ions. Although both the PAW and PA-Mg-W had antifungal effects on C. pseudolongus, their effectiveness differed, with cell viability higher in the PA-Mg-W than in the PAW. This study demonstrates that the antifungal effect of PAW could be manipulated using nitrate capture. The wide use of plasma therapy for problematic fungus control is challenging because fungi have rigid cell wall structures in different fungal groups.