Identification of a polysaccharide produced by the pyruvate overproducer Candida glabrata CCTCC M202019

• Published in: Applied microbiology and biotechnology 2017-06, Vol.101 (11), p.4447-4458
• Main Authors: Luo, Zhengshan, Liu, Song, Du, Guocheng, Zhou, Jingwen, Chen, Jian
• Format: Article
• Language: English
• Subjects: Analysis; Biomedical and Life Sciences; Biosynthesis; Biotechnological Products and Process Engineering; Biotechnology; Candida glabrata - drug effects; Candida glabrata - genetics; Candida glabrata - metabolism; Chemical analysis; Chemical bonds; Conversion; Conversion ratio; Fermentation; Flasks; Fourier transforms; Fructose; Fungi; Genes; Glucose; Glucose - metabolism; Infrared analysis; Life Sciences; Mannose; Mass distribution; Microbial Genetics and Genomics; Microbiology; Monosaccharides - chemistry; Mutagenesis; Physiological aspects; Polysaccharides; Polysaccharides - biosynthesis; Polysaccharides - chemistry; Polysaccharides - isolation & purification; Protein binding; Proteins - metabolism; Pyruvic acid; Pyruvic Acid - metabolism; Saccharides; Sugar; Temperature effects; Transcription; Transcription (Genetics); Vitamins - pharmacology
• ISSN: 0175-7598; 1432-0614
• DOI: 10.1007/s00253-017-8245-1

Candida glabrata has great potential for the accumulation of pyruvate as a preferred strain in pyruvate production by fermentation. However, its substrate conversion rate is relatively low. In this study, a novel polysaccharide containing α-1,4-glucosidic bonds was observed accidentally in screening a high-titer pyruvate strain by atmospheric and room temperature plasma mutagenesis of C. glabrata . Chemical analysis of the partially purified polysaccharide S 4-C10 showed the main components were 1.2% ( w / w ) protein and 94.2% ( w / w ) total sugar. Fourier transform infrared and molecular mass distribution analysis indicated that the main component (PSG-2) of S 4-C10 was a small molecular homogeneous protein-bound polysaccharide. Monosaccharide analysis of PSG-2 showed it consisted of glucose, mannose, and fructose. By optimizing the vitamin mix content, 77.6 g L −1 S 4-C10 polysaccharide could be obtained after 72 h fermentation at 30 °C in 500-mL flasks. RT-qPCR analysis showed that transcriptional level of some key genes related to polysaccharide biosynthesis was upregulated compared to that of wild-type strain. By knocking out two most significantly upregulated genes, CAGL0H02695g and CAGL0K10626g, in the wild-type strain, the pyruvate consumption rate was significantly reduced in late pyruvate fermentation phase, while the titer of polysaccharides was reduced by 18.0%. Besides the potential applications of the novel identified polysaccharide, this study provided clues for increasing the conversion ratio of glucose to pyruvate in C. glabrata by further decreasing the accumulation of polysaccharides.