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  • Author or Editor: S.-Q. Liu x
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Saccharomyces cerevisiae MERIT.ferm was used as mono- and mixed-cultures with Williopsis saturnus var. mrakii NCYC500 in mango wine fermentation. A ratio of 1:1000 (Saccharomyces:Williopsis) was chosen for mixed-culture fermentation to enable longer persistence of the latter. The monoculture of S. cerevisiae and mixed-culture was able to ferment to dryness with 7.0% and 7.7% ethanol, respectively. The monoculture of W. mrakii produced 1.45% ethanol. The mango wines fermented by S. cerevisiae alone and the mixed-culture were more yeasty and winey, which reflected their higher amounts of fusel alcohols, ethyl esters and medium-chain fatty acids. The mango wine fermented by W. mrakii alone was much less alcoholic, but fruitier, sweeter, which corresponded to its higher levels of acetate esters.

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Abstract

Nattokinase (NK) is effective in the prevention and treatment of cardiovascular disease. Cucumber is rich in nutrients with low sugar content and is safe for consumption. The aim of this study was to construct a therapeutic cucumber that can express NK, which can prevent and alleviate cardiovascular diseases by consumption. Because the Bitter fruit (Bt) gene contributes to bitter taste but has no obvious effect on the growth and development of cucumber, so the NK-producing cucumber was constructed by replacing the Bt gene with NK by using CRISPR/Cas9. The pZHY988-Cas9-sgRNA and pX6-LHA-U6-NK-T-RHA vectors were constructed and transformed into Agrobacterium tumefaciens EHA105, which was transformed into cucumber by floral dip method. The crude extract of NK-producing cucumber had significant thrombolytic activity in vitro. In addition, treatment with the crude extract significantly delayed thrombus tail appearance, and the thrombin time of mice was much longer than that of normal mice. The degrees of coagulation and blood viscosity as well as hemorheological properties improved significantly after crude extract treatment. These findings show that NK-producing cucumber can effectively alleviate thrombosis and improve blood biochemical parameters, providing a new direction for diet therapy against cardiovascular diseases.

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