Exploring the diversity of bacteriophage specific to Oenococcus oeni and Lactobacillus spp and their role in wine production.

The widespread existence of bacteriophage has been of great interest to the biological research community and ongoing investigations continue to explore their diversity and role. They have also attracted attention and in-depth research in connection to fermented food processing, in particular from the dairy and wine industries. Bacteriophage, mostly oenophage, may in fact be a 'double edged sword' for winemakers: whilst they have been implicated as a causal agent of difficulties with malolactic fermentation (although not proven), they are also beginning to be considered as alternatives to using sulphur dioxide to prevent wine spoilage. Investigation and characterisation of oenophage of Oenococcus oeni, the main species used in winemaking, are still limited compared to lactococcal bacteriophage of Lactococcus lactis and Lactiplantibacillus plantarum (formally Lactobacillus plantarum), the drivers of most fermented dairy products. Interestingly, these strains are also being used or considered for use in winemaking. In this review, the genetic diversity and life cycle of phage, together with the debate on the consequent impact of phage predation in wine, and potential control strategies are discussed. KEY POINTS: • Bacteriophage detected in wine are diverse. • Many lysogenic bacteriophage are found in wine bacteria. • Phage impact on winemaking can depend on the stage of the winemaking process. • Bacteriophage as potential antimicrobial agents against spoilage organisms.

Influences of light on growth, reproduction and hypocrellin production by Shiraia sp. SUPER-H168.

Light is a very important signal for fungi since it influences many different physiological responses. The effects of dark or light at different wavelengths on growth, reproduction and hypocrellins of Shiraia sp. SUPER-H168 were studied: dark, white, red, yellow, green, blue and purple. All incubations under different light conditions had significant stimulating effects on aerial hyphae and suppressing effects on hypocrellin biosynthesis compared with dark incubation. Under blue and purple light especially blue light, the colonies with profuse growth of aerial mycelium were formed. Hypocrellin production reached 13.73 mg per dish under dark condition, and decreased to 4.01 mg and 2.83 mg per dish under white and blue light, respectively. Light condition not only influenced hypocrellin production but also influenced the composition of hypocrellins. Four types of hyphae, namely surface, aerial, biofilm and penetrative hyphae, were observed by light microscopy and SEM. This study found that biofilm hyphae was so closely connected with production of secondary metabolites, and hypocrellins were only produced by biofilm hyphae. Light promoted sexual development and inhibited asexual reproduction, especially blue light strongly inhibited asexual development.

Enhanced hypocrellin production of Shiraia sp. SUPER-H168 by overexpression of alpha-amylase gene.

Relative expression levels of twenty-four amylase genes in Shiraia sp. SUPER-H168 were investigated by real-time quantitative PCR when various carbohydrates, including glucose, sucrose, maltose, amylose, amylopectin and corn flour, were used as carbon source. Genes, including an α-glucosidase gene Amy33 (2997 bp), an α-amylase gene Amy365-1 (1749 bp) and a glycogen debranching enzyme gene Amy130 (2487 bp), were overexpressed, and four overexpression transformants were constructed, respectively. When Amy365-1 and Amy130 were co-overexpressed, relative expression levels of seven hypocrellin biosynthesis genes and four related genes in central carbon catabolism were all increased. Expression of Amy33 was also increased along with increase of Amy365-1 and Amy130. Under liquid state fermentation, biomasses and hypocrellin productions were both gradually increased in four overexpression strains than those of wild type strain. Under SSF, hypocrellin production of Amy365-1 and Amy130 co-expression strain reached 71.85 mg/gds, which was 2.83 fold than that of wild type strain, and residual sugar was decreased from 35.47% to 16.68%. These results can provide a practical approach for other secondary metabolites by filamentous fungi under SSF when raw starch material is used as carbon source.