Project description:Cyberlindnera jadinii yeast is a potential sustainable novel feed ingredient for aquaculture industries. Yeasts contain bio-active components and proteins such as beta-glucans, mannans, nucleic acids and proteins that can enhance fish immunity against the disease. In our study, we focused on the characterization of intestinal immunoregulatory pathways in zebrafish (Danio rerio) by quantifying the intestine proteins with isobaric tags for relative and absolute quantitation (iTRAQ) and 2D LC-MS/MS approach. Zebrafish were fed either a control diet (C) or a diet supplemented with autolyzed C. jadinii (ACJ). The KEGG pathways analysis revealed that compared with the control diet, the ACJ yeast diet induced an increased abundance of proteins related to arginine and proline metabolism, phagosome, C-lectin receptor signalling pathway, ribosome pathway and PPAR signalling pathway, which can modulate and enhance the innate response of zebrafish. Moreover, fish fed ACJ yeast diet also showed decreased abundance of proteins associated with inflammatory pathways including apoptosis, necroptosis and ferroptosis pathways. These findings support a mobilization of the innate immune response and a control of inflammatory-related pathways in the intestine of zebrafish. Our findings in the well annotated proteome of zebrafish enabled a detailed investigation of intestinal responses and provide insight into the health-beneficial effects of the yeast species C. jadinii relevant for aquaculture species.
Project description:Cyberlindnera jadinii is widely used as a source of single-cell protein and is known for its ability to synthesize a great variety of valuable compounds for the food and pharmaceutical industries. Its capacity to produce compounds such as food additives, supplements, and organic acids, among other fine chemicals, has turned it into an attractive microorganism in the biotechnology field. In this review, we performed a robust phylogenetic analysis using the core proteome of C. jadinii and other fungal species, from Asco- to Basidiomycota, to elucidate the evolutionary roots of this species. In addition, we report the evolution of this species nomenclature over-time and the existence of a teleomorph (C. jadinii) and anamorph state (Candida utilis) and summarize the current nomenclature of most common strains. Finally, we highlight relevant traits of its physiology, the solute membrane transporters so far characterized, as well as the molecular tools currently available for its genomic manipulation. The emerging applications of this yeast reinforce its potential in the white biotechnology sector. Nonetheless, it is necessary to expand the knowledge on its metabolism, regulatory networks, and transport mechanisms, as well as to develop more robust genetic manipulation systems and synthetic biology tools to promote the full exploitation of C. jadinii.
Project description:The yeast Cyberlindnera jadinii has great potential in the biotechnology industry due to its ability to produce a variety of compounds of interest, including carboxylic acids. In this work, we identified genes encoding carboxylate transporters from this yeast species. The functional characterization of sixteen plasma membrane carboxylate transporters belonging to the AceTr, SHS, TDT, MCT, SSS, and DASS families was performed by heterologous expression in Saccharomyces cerevisiae. The newly identified C. jadinii transporters present specificity for mono-, di-, and tricarboxylates. The transporters CjAto5, CjJen6, CjSlc5, and CjSlc13-1 display the broadest substrate specificity; CjAto2 accepts mono- and dicarboxylates; and CjAto1,3,4, CjJen1-5, CjSlc16, and CjSlc13-2 are specific for monocarboxylic acids. A detailed characterization of these transporters, including phylogenetic reconstruction, 3D structure prediction, and molecular docking analysis is presented here. The properties presented by these transporters make them interesting targets to be explored as organic acid exporters in microbial cell factories.
Project description:A strand-specific RNA-seq library was constructed for a single sample, specifically the wild-type strain NBRC0988 grown in YEP medium supplemented with 2% w/v glycerol. To prepare the RNA sample, NBRC0988 cells grown overnight were inoculated into 100 mL of liquid Yeast Extract Peptone Dextrose (YEPD) medium, with an initial inoculation OD600 of 0.1. These cells were then cultured for 15 hours at 30°C and 250 rpm. Subsequently, the cells were collected by centrifugation at 6,000g for 5 minutes. After being washed twice with phosphate buffer saline (PBS), they were inoculated into fresh 100 mL of YEP medium containing 2% w/v glycerol. Following flask culturing at 30°C and 250 rpm for an additional 5 hours, the yeast cells were collected by centrifugation for total RNA isolation and Illumina RNA-seq library construction. Total RNA from the sample was isolated using the TRIzol reagent (Invitrogen, Grand Island, USA) and then utilized for high-throughput RNA sequencing. Commercially, the 150-nt paired-end strand-specific RNA-seq libraries (SS_lib_type RF) were generated by Novogene Biotechnology Co. Ltd (Tianjin, China) using the Illumina's Novaseq 6000 platform (Illumina, San Diego, USA).